Biophotonics201111 DL

November/December 2011
P
h
o
t
o
c
o
a
g
u
l
a
t
i
o
n

I
n
v
e
r
t
e
d

F
l
u
o
r
e
s
c
e
n
c
e

M
i
c
r
o
s
c
o
p
y

F
o
r
e
n
s
i
c

M
i
c
r
o
s
c
o
p
y
N
/
D

1
1
Worldwide Coverage: Optics, Lasers, Imaging, Fiber Optics, Electro-Optics, Photonics Component Manufacturing
www.Photonics.com
THE EMERGENCE OF MULTIMODAL NLO IMAGING
Cover1_Layout 1 11/1/11 9:56 AM Page 1
Bright Ideas in Fiberoptics
FOLLOW
THE LEADER
PH 508-909-2200 WWW.I NCOMUSA.COM SALES@I NCOMUSA.COM
Well help you do things you havent even thought of yet.
Medical
Life Sciences Display Scientic
Incom is the commercial leader in fused ber optics.
Incoms ber optic
faceplates continue to lead
the digital revolution in
medical X-ray technology.
Incoms ber optic tapers
help propel researchers and
equipment manufacturers
to the limits of scientic
discovery.
Incoms microwell and
microcapillary arrays
provide high-speed
solutions for genetic
sequencing in advanced
genome studies.
Incoms ber optics enable
seamless and dynamic
digital displays with tactile
feedback.
nd11_Incom_PgCVR2_Layout 1 10/27/11 10:30 AM Page CVR2
nd11_Newport_InSight_Pg3_Layout 1 10/27/11 10:32 AM Page 3
4
www.photonics.com
10 BIOSCAN
BioPhotonics editors curate the most significant headlines
of the month for photonics in the life sciences and take
you deeper inside the news. Featured stories include:
DNA nanosensors pave way for cancer tests, drugs
STED microscopy reveals how immune system
attacks infected cells
Laser technique produces synthetic tissue
for regenerative medicine
21 BUSINESSSCAN
Photonics-related projects score NIH grants
NEWS
35
BioPhotonics November/December 2011
Volume 18 Issue 9
t
TABLE OF CONTENTS
24 TO LABEL OR NOT?
by Dr. Neil Anderson, Semrock Inc., a unit of Idex Corp.
The emerging field of multimodal nonlinear optical imaging, which
integrates label-based and label-free methods, could become the
definitive diagnostic tool of the future.
28 PHOTOCOAGULATION: HEALING THROUGH LASER DAMAGE
by Marie Freebody, Contributing Editor
Used in almost all surgical specialties, this decades-old method of containing
tissue damage has become especially important to ophthalmologists.
31 DIFFRACTION-IMAGING FLOW CYTOMETRY ENABLES RAPID CELL ASSAY
by Xin-Hua Hu, East Carolina University and Wavmed Technologies Corp.;
Yuanming Feng, Tianjin University; and Jun Qing Lu, East Carolina University
A novel method of combining flow cytometry with diffraction rather than fluores-
cence imaging enables 3-D visualization and suggests new applications in medicine.
35 CSI EXPERTS FIND CLUES FASTER WITH MICROSCOPY
by Marie Freebody, Contributing Editor
The field of forensics is being catalyzed by the efficiencies of digital imaging
and by combining optical techniques with spectroscopy and spectrophotometry.
38 INVERTED FLUORESCENCE MICROSCOPY AIDS MICROSEPARATION STUDIES
by Dr. Yolanda Fintschenko, Labsmith Inc., and Dr. Blanca Lapizco-Encinas,
Tennessee Technological University
This microscope has the stationary stage and software required by researchers
working with micro- or nanofluidic experiments.
FEATURES
NEWS
7 EDITORIAL
8 LETTERS
43 BREAKTHROUGHPRODUCTS
48 APPOINTMENTS
Upcoming Courses and Shows
49 ADVERTISER INDEX
50 POST SCRIPTS
by Laura S. Marshall
Genetic approach shows bright promise against AIDS
DEPARTMENTS
PHOTONICS
The technology of generating and harnessing light and other forms of radiant energy whose
quantum unit is the photon. The range of applications of photonics extends from energy generation
to detection to communications and information processing.
BIOPHOTONICS
The application of photonic products and techniques to solve problems for researchers,
product developers, clinical users, physicians and others in the fields of medicine,
biology and biotechnology.
This months cover is based
on an article about multimodal
nonlinear optical imaging.
Written by Dr. Neil Anderson
of Semrock Inc., it begins
on page 24. Design by Art
Director Suzanne L. Schmidt.
ND11_Contents_Layout 1 10/27/11 11:57 AM Page 4
nd11_OptBuildingBlocks_Pg5_Layout 1 10/27/11 10:33 AM Page 5
www.photonics.com
Group Publisher Karen A. Newman
Editorial Staff
Managing Editor Laura S. Marshall
Senior Editor Melinda A. Rose
Features Editor Lynn M. Savage
News Editors Gary Boas, Caren B. Les, Ashley N. Paddock,
Krista Zanolli
Contributing Editors Hank Hogan, Marie Freebody
Copy Editors Judith E. Storie, Patricia A. Vincent,
Margaret W. Bushee
Creative Staff
Senior Art Director Lisa N. Comstock
BioPhotonics Art Director Suzanne L. Schmidt
Designer Janice R. Tynan
Director of Publishing Operations Kathleen A. Alibozek
Electronic Media Staff
Director Charley Rose
Multimedia Services & Marketing
.NET Developers Brian L. LeMire, Alan W. Shepherd
Corporate Staff
Chairman/CEO Teddi C. Laurin
President Thomas F. Laurin
Director of Sales Ken Tyburski
Controller Mollie M. Armstrong
Accounting Manager Lynne Lemanski
Accounts Receivable Manager Mary C. Gniadek
Business Manager Elaine M. Filiault
Human Resources Coordinator Carol J. Atwater
Business Staff
Advertising Production Coordinator Kristina A. Laurin
Trade Show Coordinator Allison M. Mikaniewicz
Computer Systems Manager Deborah J. Lindsey
Computer Assistant Angel L. Martinez
Circulation Manager Heidi L. Miller
Assistant Circulation Manager Melissa J. Liebenow
Circulation Assistants Alice M. White, Kimberly M. LaFleur,
Theresa A. Horn
Subscriptions Janice L. Butler
Distribution Manager George A. Houghtlin
Traffic Manager Daniel P. Weslowski
EDITORIAL MAIN OFFICE
Laurin Publishing, Berkshire Common, 2 South St.
PO Box 4949, Pittsfield, MA 01202-4949
+1 (413) 499-0514; fax: +1 (413) 442-3180; e-mail: editorial@photonics.com
Subscription Policy BioPhotonics ISSN-1081-8693, (USPS 013913) is published 9 times per year by Laurin
Publishing Co. Inc. TITLE reg. in US Library of Congress. The issues will be as follows: January, February,
March, April, May/June, July/August, September, October, November/ December. Copyright 2011 by Lau-
rin Publishing Co. Inc. All rights reserved. POSTMASTER: Periodicals postage paid at Pittsfield, MA, and at ad-
ditional mailing offices. Postmaster: Send form 3579 to BioPhotonics, Berkshire Common, PO Box 4949,
Pittsfield, MA 01202-4949, +1 (413) 499-0514. CIRCULATION POLICY: BioPhotonics is distributed without charge
to qualified researchers, engineers, practitioners, technicians and management personnel working with the
fields of medicine or biotechnology. Eligibility requests must be returned with your business card or organi-
zations letterhead. Rates for others as follows: $45 domestic and $56.25 outside US per year prepaid. Over-
seas postage: $30 airmail per year. Publisher reserves the right to refuse nonqualified subscriptions. ARTICLES
FOR PUBLICATION: Individuals wishing to submit an article for possible publication in BioPhotonics should
contact Laurin Publishing Co. Inc., Berkshire Common, PO Box 4949, Pittsfield, MA 01202-4949; phone: +1 (413)
499-0514; fax: +1 (413) 442-3180; e-mail: editorial@ photonics.com. Contributed statements and opinions ex-
pressed in BioPhotonics are those of the contributors the publisher assumes no responsibility for them.
6 BioPhotonics November/December 2011
ND11_Masthead_Layout 1 10/27/11 10:10 AM Page 6
Enhance, Disrupt, Revolutionize, Repeat
I
n his introduction to a plenary session called Material
Nanoscience, Photonics and Technologies for Revolutionary
Innovation at LIAs ICALEO (International Congress on
Applications of Lasers & Electro-Optics) last month in Orlando,
Fla., session chairman Kunihiko Washio of Paradigm Laser
Research Ltd. in Tokyo said, Although incremental innovation
within a known systematic framework is very important for
steady technology progress, disruptive innovation is often desired
to push the limits of the imaginable and to leap forward to attain
revolutionary innovation.
Disruptive innovator Steve Jobs, the former Apple CEO,
changed the world with his computers and phones, and just one
day before he died last month at the age of 56, a press release
from the University of California, Davis, described an adaptation
to his iPhone that turned it into a microscope capable of both
revealing vital medical information and transmitting images for
analysis and diagnosis. I wonder if Jobs knew about the adapta-
tion. I wonder what he thought, if he knew, about his disruptive
phone being turned into a potentially game-changing device to
bring a new level of health care to the world.
The group at UC-Davis was not the first to build a smartphone
microscope, and it certainly wont be the last to add some good,
incremental innovation to an already great idea, whatever it may
be. And thats good for all of us. (You can read more about the
UC-Davis enhanced iPhone on photonics.com, http://www.
photonics.com/a48604.) Innovation of all kinds keeps an industry
growing, and thats pretty good, too.
In his introduction at ICALEO, Washio went on to say, Mate-
rials science, particularly material nanoscience, is believed to be
the treasure house of the seeds of desired disruptive innovations.
The keynote presentation that followed, The Story and Prospects
of Carbon Nanoscience and Technologies for Future Exciting
Applications by Stanford researcher Hongjie Dai, presented a
look at carbon nanotubes and their unique intrinsic physical
and chemical properties, which can be exploited for biological
and biomedical applications including detection, diagnostics,
imaging and novel therapy.
We cant wait to see where the next disruptive and incremental
biophotonics innovations will come from. In a small change of
our own, were going to focus a couple of articles in every issue
around a specific topic, exploring aspects of the subject that are
bringing real change to the way we think about biophotonics,
its applications and our world.
You might see an update on the smartphone microscope in
our September issue coverage of biophotonics and global health,
while Dais work on carbon nanotubes may make its way into
our March issue, with its focus on biophotonics and cancer. With
so much interesting work going on in photonics and the life
sciences, we wanted to organize it and bring it to you with some
sense of the impact it could have on the way we live. Well ex-
plore photonics in dentistry in January. In February, well take a
closer look at biophotonics in the pharmaceutical industry. Other
topics will be covered, too, and all the things you like about Bio-
Photonics will still be there, just enhanced. We hope you like it.
In the meantime, enjoy this issue, and make plans to visit us
at our booths at BiOS (8327) and Photonics West (323) in San
Francisco in January.
7
EDITORIAL
Karen A. Newman
karen.newman@photonics.com
ND11_Editorial_Layout 1 10/27/11 10:33 AM Page 7
8
The limits of diffraction limits
In your recent article on high-resolution
microscopy (July/August 2011, p. 31),
the heading asks, Can optical micros-
copy further shred the Abbe diffraction
limit? Although the statement about
shredding the Abbe diffraction limit often
is perpetuated by those working in the
field of STED [stimulated emission
depletion] microscopy, it is basically
wrong. The Abbe resolution criterion
(or Rayleigh, for that matter) is valid only for real imaging
systems like a microscope lens. It basically teaches us that, even
if the best optical materials are used in the lens manufacturing,
the resolution of the lens will be limited by diffraction of the
lens aperture.
The new, and impressive, scanning techniques like STED and
PALM [photoactivated localization microscopy] rely on a clever
interplay between fluorophores and sophisticated laser scanning
methods and, as such, are more related to raster scanning known
in scanning electron microscopes.
It is my hope that the erroneous comparison of classical imag-
ing and laser scanning methods will be avoided in the future.
Lars Ren Lindvold
Senior Scientist, Radiation Research Div.
Ris National Laboratory for Sustainable Energy
Technical University of Denmark
Letters
From UV to NIR, Hamamatsus extensive family of scientic cameras have your imaging
needs covered. We offer over 60 high-quality cameras including our ve major cameras: a
high-speed scientic CMOS camera, a versatile cooled CCD camera, a dual-CCD camera
for dual wavelength imaging, and ultra sensitive EMCCD cameras.
For complete specications of our major cameras, go to http://hamamatsucameras.com.
Learn more about our family of products at
www.hamamatsucameras.com, or call 1-800 524 0504.
NEED A
CAMERA?
HTTP://SALES.HAMAMATSU.COM
USA 1-800 524 0504, FREEPHONE EUROPE 00 800 800 800 88
ORCA-D2
ImagEM
ORCA-Flash2.8
ORCA-R2
ND11_Letters_Layout 1 10/27/11 10:11 AM Page 8
Photonics Medias industry-leading site features the latest industry news and events
from around the world.
Welcome to
Web Exclusive:
Optogenetics using light to control geneti-
cally altered living cells began as a way
to probe neural code and reveal the secrets
of disorders in the still-mysterious human
brain. But teams at two universities are taking
their optogenetic work to heart, literally,
with the goal of replacing electrical pace-
makers. News Editor Caren Les asked
Stanfords Oscar Abilez and Stony Brook
University Medical Centers Emilia Entcheva
about the challenges they face and whats
next for their research. For more, visit:
www.photonics.com/webexclusives
2011 Prism Awards Finalists
Finalists for the 2011 Prism Awards for photonic innovation have
been chosen! Upon reviewing nearly 150 cutting-edge product
entries from around the world, our prestigious panel of judges
has selected the best and brightest in the industry. Winners will
be announced on Jan. 25, 2012, at SPIE Photonics West. For a
list of finalists, visit: Photonicsprismaward.com
Sponsored by
Search Biophotonics-Related News
nd11Online_Layout 1 10/27/11 10:12 AM Page 9

BIOSCAN
SANTA BARBARA, Calif. Custom
DNA molecules can be used to make sen-
sors that can quickly detect a broad class
of proteins and could be used to person-
alize cancer treatment and even to monitor
the quality of stem cells.
Researchers from the University of Cal-
ifornia, Santa Barbara (UCSB), and the
University of Rome Tor Vergata developed
the new nanosensors, which monitor the
activity of proteins called transcription
factors, then read the genome and translate
it into instructions for synthesizing the
various molecules that compose and con-
trol the cell. This information could deter-
mine which transcription factors in a pa-
tients cancer cells are activated or
repressed, enabling physicians to prescribe
the right combination of drugs for each
patient, according to Alexis Valle-Blisle,
a postdoctoral researcher in UCSBs de-
partment of chemistry and biochemistry.
Andrew Bonham, a postdoctoral scholar
at UCSB and co-first author of the study,
explains that the teams approach to read-
ing transcription factors is quick and con-
venient. By adding their sensors to the
mashed-up cells, they can measure the
samples level of fluorescence.
The international research effort or-
ganized by senior authors Kevin Plaxco
and Francesco Ricci started when Ricci
realized that all of the information neces-
sary to detect transcription factor activities
is already encrypted in the human genome
and could be used to build sensors. Once
activated, each of the thousands of differ-
ent transcription factors can bind to its
own specific target DNA sequence, he ex-
plained. These sequences were used as the
foundation for building the nanosensors.
The key to the technology came from
studies of the natural biosensors inside
cells. The scientists expounded upon the
fact that all creatures monitor their sur-
roundings with biomolecular switches
composed of RNA or proteins, which are
small enough to operate inside a cell and
could work in complex environments. In-
spired by the efficiency of these natural
nanosensors, the researchers teamed with
professor Norbert Reich of UCSB to build
synthetic switching nanosensors using
DNA rather than proteins or RNA.
Specifically, they re-engineered three
naturally occurring DNA sequences, each
recognizing a different transcription factor,
into molecular switches that become fluo-
rescent when they bind to their intended
targets. Using these nanosensors, the re-
searchers can determine transcription fac-
tor activity directly in cellular extracts by
measuring their fluorescence level. The
work was described in an online article
published Aug. 4 in the Journal of the
American Chemical Society (doi: 10.1021/
ja204775k).
This strategy ultimately will allow biol-
ogists to monitor the activation of thou-
sands of transcription factors, leading to a
better understanding of the mechanisms
underlying cell division and development.
The nanosensors could also be used to
screen and test new drugs that could in-
hibit the transcription factor binding activ-
ity responsible for tumor cell growth.
DNA nanosensors pave way for cancer tests, drugs
A structure-switching nanosensor made from DNA (blue and purple) detects a specific transcription factor
(green). Using these nanosensors, researchers have demonstrated the direct detection of transcription factors in
cellular extracts. They believe that their strategies will allow biologists to monitor the activity of thousands of
transcription factors and will lead to more efficient cancer testing and medications. Courtesy of Peter Allen.
Alexis Valle-Blisle (left) and Andrew Bonham
(right). Courtesy of George Foulsham, Office of
Public Affairs, UCSB.
ND11_BioScan_Layout 1 10/27/11 2:27 PM Page 10
BioPhotonics November/December 2011 11
STED microscopy reveals how immune system attacks infected cells
PHILADELPHIA A new stimulated
emission depletion (STED) microscope
has yielded unprecedented views of the
immune system in action.
The superresolution microscope shows
how granules from natural killer (NK)
cells pass through openings in the dy-
namic cell skeleton to destroy their tar-
gets: tumor and virus-infected cells. Un-
derstanding these biological events better
may soon allow researchers to devise
more effective treatments for inherited
diseases that leave the immune system
compromised.
From a biological perspective, the
work defines a previously unappreciated
regulatory hurdle that an NK cell must
overcome in order to access essential host
defense functions, said Dr. Jordan S.
Orange of the Childrens Hospital of
Philadelphia. This of course presents
opportunities for exploiting this process
in order to obtain more or less of this par-
ticular variety of secretion. This could
have very relevant clinical and therapeutic
implications.
Previously, conventional fluorescence
light microscopes could not resolve ob-
jects smaller than 200 nm. The new STED
technique uses a unique arrangement of
lasers and fluorescence to image fine
structures such as protein filaments
smaller than 60 nm.
Orange has long researched the biology
of NK cells at the immunological synapse
the site where the NK cell attaches to its
target cell and delivers cell-killing mole-
cules. A crucial component of this highly
regulated process is filamentous actin
(F-actin), a protein in NK cells that forms
a dense network through which cell-killing
molecules move into the synapse.
It was thought that F-actin is not present
at the center of the network, where the
granules fuse with the cell surface. The
current study reveals under superresolu-
tion, however, that F-actin pervades the
synapse but leaves openings just large
enough to allow granules to pass through.
The scientists observed that F-actin ap-
peared to be dynamically interacting with
the granules to move them toward their
targets.
Orange compared the F-actin filaments
to the rails of a roller coaster that quickly
rearranges itself to guide a rider through a
narrow tunnel. He explained that further
studies of NK function will investigate en-
ergy use and biological mechanisms that
allow the lytic granules to navigate the im-
munological synapse.
The patterning and coordination of the
pervasive actin network is telling a story
one that is likely to underscore how the
critical host defense function mediated by
NK cells is accessed, Orange said. We
certainly need to sort this out as well as
what underlies the interaction between
NK cell lytic organelles and hypodense
regions within the pervasive actin at the
synaptic interface.
The study was published Sept. 13 in the
online open-access journal PLoS Biology
(doi: 10.1371/journal.pbio.1001151).
Platinum rotary electron micrograph of a natural
killer cell cortex colorized to depict actin filaments
(blue) and a single intercalated lytic granule
(yellow). Courtesy of Dr. Jordan Orange.
Superresolution stimulated emission depletion (STED)
fluorescence micrograph of an activated human
natural killer cell demonstrating actin filaments
(green) and perforin-containing lytic granules
(red). Courtesy of Dr. Emily Mace.
AACHEN, Germany Tissue
that has been damaged by dis-
ease or trauma often cannot
repair itself. However, a new
picosecond laser technique pro-
duces biomimetic matrices that
allow the body to regenerate it-
self using the patients own
cells.
The process was developed
by researchers at the Fraunhofer
Institute for Laser Technology
ILT and other Fraunhofer insti-
tutes. The biomimetic scaffolds
closely emulate endogenous tis-
sue and enable fabrication of
specialized model systems for
studying 3-D cell growth. The
researchers combined organic
substances with polymers to
produce 3-D structures that are
suitable for building artificial
tissue.
Tissue engineering is a
highly interesting and versatile
research area with huge appli-
cation potential, said Sascha
Engelhardt, project manager at
ILT. However, scaffolds devel-
oped for tissue engineering are
not yet fully able to mimic their
complex natural models.
Normally, scaffold produc-
tion is focused either on
structural, mechanical or bio-
chemical aspects, but Engel-
hardt explained that all three
must be considered in a single
scaffold because they all have
been shown to influence cell
behavior significantly.
By controlling structure,
mechanics and biochemistry in
a single experiment, the com-
plexity of the natural model can
Laser technique produces synthetic tissue for regenerative medicine
Capillaries of an artificial resilient poly-
mer with a diameter of 20 m. Images
courtesy of Fraunhofer Institute for
Laser Technology ILT.
EDMONTON, Alberta, Canada The
palette of fluorescent highlighters used
to track the movement of messengers in-
side single cells has been dramatically ex-
panded to include red and blue fluorescent
indicators, which provide researchers a
vivid full-color view of calcium ions mov-
ing about in their role as key intracellular
signaling messengers.
Until recently, cellular imaging of the
calcium ion required the use of a green
fluorescent indicator. Accordingly, imag-
ing of calcium ions produced monochro-
matic images and movies in shades of
green. Now, scientists at the University of
Alberta have added red and blue indica-
tors.
The well known selection of fluores-
cence protein colors has proved useful for
live-cell imaging of multiple organelles
and proteins, according to doctoral candi-
date Yongxin Zhao and Robert Campbell,
associate professor of chemistry. They ex-
plained that a similar palette of calcium
indicators will enable researchers to start
designing experiments that involve visual-
izing multiple dynamic biochemical pa-
rameters simultaneously.
Imaging of the calcium ion is com-
monly used by researchers to monitor cel-
lular activity such as the firing of neurons.
However, since calcium ions are colorless,
it is necessary to introduce colored indica-
tor proteins into the cell, which can either
increase in fluorescent brightness or
change the fluorescence color when they
bind to the ions. These changes in bright-
ness can be visualized easily using appro-
priate microscopy equipment.
Examining the dynamics of calcium
ions inside a single cell in better detail
could help pharmaceutical researchers de-
termine whether a drug designed to affect
a specific cell is hitting its target. In addi-
tion, it could help scientists better visual-
ize neuronal activity in model organisms
such as transgenic worms or mice.
The team, led by Zhao, engineered the
new genetically encoded indicator proteins
using bacterial cells. The indicator genes
b
BIOSCAN
be reduced to its essentials, he
said. From this gained knowl-
edge, a translation in a
technical solution will be
more feasible.
The researchers used dis-
solved endogenous proteins
such as albumin, collagen and
fibronectin and polymers irra-
diated with laser light and cross-
linked by photolytic processes.
To achieve this, they deployed
picosecond laser pulses from a
low-cost microchip laser to trig-
ger a multiphoton polymeriza-
tion process. The short pulse
duration caused almost no heat
damage to the material. Ultra-
fast megawatt-range pulses
drove a massive number of
protons into the laser focus in
an extremely short time, trigger-
ing a nonlinear effect.
The molecules in the liquid
absorbed several photons simul-
taneously, causing free radicals
to form. This multiphoton poly-
merization process triggered
chemical reactions between the
surrounding molecules, forming
solids from the liquid. CAD
data helped the system guide
the position of the laser beam
through a microscope with the
precision of a few hundred
nanometers in such a way that
micrometer-fine, stable volume
elements of cross-linked mate-
rial gradually formed.
With resolution of approxi-
mately 1 m, the process en-
abled the team to produce cell
scaffolds directly from dis-
solved proteins and polymers,
Engelhardt said. Offering
much greater stability, the scaf-
folds can be seeded with the
patients own cells in a med-
ical laboratory, and then the
colonized scaffolds can be ex-
pected to produce good im-
plant growth in the patients
body. The long-term aim is to
produce not only individual
cell colonies but also artificial
tailor-made organs.
Our next aim will be to de-
sign and develop an in vitro
assay in the form of a con-
trolled 3-D microenvironment
based on our technology, En-
gelhardt said. Of course, this
assay will not be realized alone
by our institute, but within an
interdisciplinary cooperation
of research partners. Our long-
term goal is to upscale our
approach by increasing the
process speed.
To reduce the time and
cost of producing tailor-made
supporting structures for syn-
thetic tissue, the team wants
to combine its fabrication
process with other rapid proto-
typing methods.
Test matrix consisting of a polymer
support structure and a protein
functional structure.
Vibrant palette of fluorescent highlighters tracks cells
Multicolor imaging with GECOs. HeLa cells transfected with nucleus-localized R-GECO1, cytoplasmic
G-GECO1, and mitochondria-localized GEM-GECO1. (Top left) Red fluorescence. (Top right) Green
fluorescence with cyan (~470 nm) excitation. (Bottom left) Pseudocolored ratio of blue to green fluorescence
with UV (~380-nm) excitation. (Bottom right) Merge of the three images, with GEM-GECO1 ratio in magenta.
were programmed to be sent to the outside
of the bacterial cytoplasmic membrane.
The calcium ion concentration then could
be experimentally altered to find the indi-
cator variants that had the desired color
and largest changes in brightness. Using
this approach, the researchers performed
directed laboratory evolution to ultimately
provide the optimized indicator proteins.
Campbell and Zhao said that their next
priority is to expand the color palette of
the indicators to include the yellow, or-
ange and far-red regions of the spectrum.
In addition, they plan to distribute the
genes to as many research groups as
possible through Addgene, a nonprofit
plasmid repository.
The research appeared online Sept. 8
in Science (doi: 10.1126/science.1208592).
b
BIOSCAN
Three-color fluorescence imaging of HeLa cells
transfected with plasmids encoding R-GECO1
(red indicator) targeted to the nucleus, G-GECO1
targeted to the cytoplasm and GEM-GECO1 to the
mitochondria. Courtesy of Robert E. Campbell et al.
New microscope delves beneath skin to detect cancer
ROME A new type of laser-scanning
confocal microscope provides more infor-
mation than previous versions and holds
promise for skin cancer diagnostics.
Unlike typical laser-scanning confocal
microscopes that take 3-D images of thick
tissue samples by visualizing thin slices
one layer at a time, the new device gathers
spectrographic information at a wide spec-
tral range approximately 0.5 to 2.5 m
for every point in the sample, and all in a
single scan. This spectroscopic fingerprint-
ing was detailed online Aug. 18 in AIP
Advances (doi: 10.1063/1.3631661).
Physicists at Consiglio Nazionale delle
Ricerche (CNR), in collaboration with a
dermatologist, demonstrated the technol-
ogy by taking high-resolution images of
the edge of a silicon wafer and of metallic
letters painted onto a piece of silicon less
than a half-millimeter wide. They also
demonstrated that it is possible to apply
this technique to a tissue sample in this
case, chicken skin without destroying it.
The main aim of our effort is not only
to produce typical histologic results with-
out tissue removal and specific prepara-
tion, but also to obtain reliable functional
An illustration of the CNR confocal microscope showing the 80-m-wide images from a silicon/silicon dioxide
calibration sample. The 10-m periodical structure is made of 100-nm-thick SiO
2
squares over a silicon
substrate. The left image is obtained with the 580-nm wavelength. The scientists can compute the full-color
image (right) by averaging the reflectivity intensities around the red, green and blue regions and associating
a suitable RGB color map. The greatest reflectivity for silicon is depicted by the green-blue portion of
the spectrum, while the greatest contribution of the SiO
2
island is depicted by the red-infrared part.
Courtesy of F.R. Bertani, L. Ferrari, S. Selci, ISC-CNR, unpublished results.
data on the tissue from spectral analysis,
said Dr. Stefano Selci of CNR. The po-
tential impact can be wide and deep.
Because the spectral region is vast,
Selci explained, the innovation could
affect many applications and contexts, in-
cluding dermatology, cosmetology, en-
doscopy for recessed diagnosis of general
tissues as well as semiconductors and any
materials science research.
With further testing, the microscope
could be used to detect early signs of
melanoma, Selci said. We are examining
now a wide range of human skin samples
ex vivo to accumulate experience on sig-
nificant samples and to correlate confocal
spectroscopy data to specific cell types
and skin structures, he said. On the tech-
nical side, we have mainly to speed up ac-
quisition times as needed for biological
specimens: Our microscope has been real-
ized from scratch to avoid any compro-
mise, and we have to invent new solutions
for our particular optical setup to transport
the brilliant white laser beam.
Until then, he said, it is suitable for
nonmedical applications such as inspect-
ing semiconductor surfaces.
b
BIOSCAN
Sensor enables cheap, portable blood testing
TOLEDO, Ohio A low-cost, portable
technique quickly and reliably detects spe-
cific proteins in a sample of human blood,
which could benefit a range of medical
sensing applications, including diagnosis
of cancer and diabetes long before the
onset of clinical symptoms.
University of Toledo scientists attached
artificially created molecules called ap-
tamers to free-floating proteins in the
blood. Aptamers are commercially avail-
able, custom-made short strands of nucleic
acid that resemble antibodies found in the
body because they connect to one type of
molecule only. Specific aptamers can be
used to search for target compounds rang-
ing from small molecules such as drugs
and dyes to complex biological molecules
such as enzymes, peptides and proteins.
However, aptamers have advantages
over antibodies in clinical testing. They
can tolerate a wide range of pH and salt
concentrations, they have high heat stabil-
ity, and they are easily synthesized and
cost-efficient. Aptamer sensors also can be
reversibly denatured, meaning that they
can easily release their target molecules,
making them perfect receptors for biosens-
ing applications.
To demonstrate the applicability of the
technique, the scientists chose thrombin
a naturally occurring protein in humans
that plays a role in clotting and throm-
bin-binding aptamers, which they attached
to a sensor surface, in this case a glass
slide coated with a nanoscale layer of
gold. As they applied the blood sample to
the testing surface, the aptamer and corre-
sponding proteins latched together.
They then used surface plasmon reso-
nance to determine whether the pairing
had been successful. If the protein was
present and had bound to the aptamer,
conditions for which resonance would
occur at the gold layer would have
changed. This change can be detected
through a simple reflectance technique
that is coupled to a linear detector.
The demonstrated surface plasmon res-
onance sensing modality is well suited for
the commercialization of portable hand-
held devices, and the aptamer-based func-
tionalization technique provides targeted
selectivity for specific protein detection,
said Dr. Brent Cameron of the department
of bioengineering at the university.
A current process based off systematic
evolution of ligands by exponential ampli-
fication (SELEX) is used by our group as
a convenient method for identifying and
optimizing unique aptamers specific to a
given target (i.e., type of protein or modi-
fied protein). Therefore, the sensing possi-
bilities are endless.
The new technique, which Cameron
said could be commercially available in
three to five years, was described in the
Aug. 31 issue of the Optical Society of
Americas open-access journal, Biomed-
ical Optics Express (doi: 10.1364/
BOE.2.002731).
Dr. Brent Cameron (right) and doctoral student Rui Zheng (left) evaluate a custom-developed
functionalized localized surface plasmon resonance microchip used in the detection and measurement
of albumin protein via a fiber optic light delivery/detection spectral probe. (Inset) A scanning electron
micrograph of the gold- based functionalized nanoparticle array used in localized surface plasmon
resonance measurements. Courtesy of Brent Cameron.
LAUSANNE, Switzerland Digital holo-
graphic microscopy (DHM) allows obser-
vation of neuronal activity in real time and
in three dimensions with up to 50 times
greater resolution than ever before
showing promise in testing new drugs to
fight neurodegenerative diseases such as
Parkinsons and Alzheimers.
DHM is a noninvasive approach that
enables extended observation of neural
processes without electrodes or dyes that
damage cells. The method yields impor-
tant information about the shape, dynam-
ics and activity of neurons, and creates
3-D navigable images down to 10-nm pre-
cision, according to senior team member
Pierre Marquet of cole Polytechnique
Fdrale de Lausanne (EPFL).
The method consists of pointing a sin-
gle-wavelength laser beam at neurons,
collecting the distorted wave on the other
side and comparing it to a reference beam.
A computer then numerically reconstructs
a 3-D image of the neurons using an algo-
rithm developed by the researchers. The
laser beam travels through the transparent
cells to obtain important information about
their internal composition.
Although DHM normally is used to de-
tect minute defects in materials, the team
decided to try using it for neurobiological
applications. In the study, the group in-
duced an electric charge in a culture of
neurons using glutamate, the main neuro-
transmitter in the brain. This charge trans-
fer carries water inside the neurons and
changes their optical properties in a way
that can be detected only by DHM. Thus,
the technique accurately visualized the
electrical activities of hundreds of neurons
simultaneously, in real time, without
harming them with electrodes, which can
record activity from only a few neurons
at a time. The groups findings appeared
in the Aug. 17 issue of The Journal of
Neuroscience (doi: 10.1523/jneurosci.
0286-11.2011).
This work has permitted us to obtain
an intrinsic optical signature of the activity
of many neurons in culture simultane-
ously, in real time and in a noninvasive
manner, Marquet said. Practically, the
high phase measurement accuracy allows
us to rapidly observe the effects of various
pharmacological substances mediated by
the activation of specific membrane recep-
tors/transponders.
Within this framework, DHM repre-
sents a very promising approach to de-
velop a unique, label-free, high-content
screening technique which is highly rele-
vant for the discovery of new drugs.
This advance in high-content screening
has important ramifications for the discov-
ery of new drugs that combat or prevent
neurodegenerative diseases, since new
molecules can be tested more quickly and
in greater numbers. The researchers have
found that lab work that previously took
12 hours can now be done in 15 to 30
minutes.
The success of DHM for high-content
screening has resulted in a startup com-
pany called Lynce Tec SA.
b
BIOSCAN
Microscopy, holography team up
to reveal neuronal activity
A 3-D representation of a
live mouse cortical neuron in
culture. Each pixel represents
a color-coded quantitative
measurement of the phase shift
induced by the neuron on the
transmitted wavefront. Courtesy
of Pierre Marquet, EPFL.
Lasers | Lenses | Mirrors | Assemblies
Windows | Shutters | Waveplates | Mounts
Americas +1 505 296 9541
Europe +31 (0)316 333041
Asia +81 3 3407 3614
AllThingsPhotonic.com
MulLimillion cycle desiqn lile
LxLremely low power consumpLion
ProprieLary lowlricLion, durable,
hiqh lR emissiviLy coaLinq
Desiqned lor exLreme vibraLion
and temperature conditions
How can we help you make your
project a success?
For more inlormaLion on
CVl Melles CrioL ShuLLers qo Lo
cvimellesqrioL.com/ShuLLers
When failure is not
an option and rapid
customization is a
requirement.
CHAMPAIGN, Ill. A new imaging
technique called spatial light interference
microscopy (SLIM) soon may answer the
much debated question as to whether
cells grow at a constant rate or exponen-
tially.
An extremely sensitive method, SLIM
can quantitatively measure mass with
femtogram accuracy using two beams of
light. It can gauge the growth of a single
cell and even mass transport within the
cell. It also can measure all cell types,
including bacteria, mammalian cells,
adherent and nonadherent cells, and popu-
lations, according to Mustafa Mir, a grad-
uate student at the University of
Illinois. The method was reported in
the July 25 issue of Proceedings of the
National Academy of Sciences (doi:
10.1073/pnas.1100506108).
Combining phase-contrast microscopy
and holography, SLIM does not require
staining or any other special preparation.
The noninvasive method uses white light
and can be used with more traditional
b
BIOSCAN
University of Illinois researchers have developed an
imaging technique called spatial light interference
microscopy (SLIM) that can quantitatively measure
cell mass with light. Courtesy of Quantitative Light
Imaging Laboratory.
Artists rendition of a SLIM measurement of a pair
of human osteosarcoma cells (real data). The colors
on the image correspond to the dry mass density at
each point. Courtesy of Quantitative Light Imaging
Laboratory and the Imaging Technology Group
Visualization Laboratory, Beckman Institute for
Advanced Science and Technology.
SLIM method measures cell growth
microscopy techniques, such as fluores-
cence, to monitor a cell as it grows.
Thanks to the methods sensitivity, re-
searchers at the university were able to
monitor cell growth through the phases
of the cell cycle. They discovered that
mammalian cells show clear exponential
growth only during the G2 phase after
the DNA replicates and before the cell di-
vides. This information has great implica-
tions not only for basic biology but also
for diagnostics, tissue engineering and
drug development.
The researchers hope to apply their new
findings on cell growth to various disease
models. For example, they plan to use
SLIM to see how growth varies between
normal cells and cancer, and to determine
the effects of treatments on growth rate.
Of all the current growth measurement
techniques, SLIM is unique in its capabili-
ties to study these interactions in typical
culture conditions, said Gabriel Popescu,
a member of the Beckman Institute for
Advanced Science and Technology at the
university. Since SLIM is designed as an
add-on module to a commercial phase mi-
croscope, it requires minimal retraining to
use and can readily be implemented in any
biology laboratory. We expect that these
capabilities will allow others to explore
other fundamental questions in the field of
cell growth.
Popescu has established SLIM as a
shared resource on the universitys cam-
pus, hoping to harness its flexibility for
basic and clinical research in a number
of areas.
Americas +1 760 438 2131
Europe +31 (0)316 333041
Asia +81 3 3407 3614
Better Stability
for Fluorescence
New solid-state lasers with
integrated ber delivery are ideal
for uorescence based applications
where stable power and beam
proles are needed for high
condence results and imaging.
Collimation optics, PM ber and
modulation (488nm) are optional.
Lgkf*'dN[\c`m\i\[Xk,-(ed
Lgkf*'dN[\c`m\i\[Xk+//ed
JkXYc\]ifd('kf+'[\^i\\j:
How can we help make
your results better?
Stable Power
Better Results
b
BIOSCAN
Cells open wide for nanothermometers
PRINCETON, N.J. Nanoscale thermometers have revealed for the first time that
individual cells inside the human body register various temperatures and do not ad-
here to the familiar 98.6 F norm.
For many years, scientists have suspected that temperatures vary inside individual
cells because many chemical reactions and physical changes take place there, pro-
ducing energy and heat. Some cells are more active than others, and the unused
energy is discharged as heat. Parts of an individual cell may be warmer because
they harbor biochemical power plants, known as mitochondria.
We have been very interested in understanding molecular reactivity inside living
cells, said Haw Yang, associate professor at Princeton University. One key aspect
in chemical reactions is temperature. Considering the highly compartmentalized and
heterogeneous nature of intracellular space, one might expect that temperature re-
sponse be nonuniform. Yet, before our work, there was no experimental evidence to
show whether it is true.
To measure the temperature of individual cells, Yang and Liwei Lin of the Univer-
sity of California, Berkeley, developed nanothermometers consisting of quantum
dots, in this case cadmium and selenium, which emit different wavelengths of light
depending on the temperature.
They inserted the dots into some mouse cells growing in lab dishes and found a
range of temperatures throughout the cell. Yang and his colleagues stimulated cellular
activity to watch the changes, reporting a difference of a few degrees Fahrenheit
in some cells. The team does not yet have enough data to give exact calculations,
however.
The temperature changes could have a major impact on how cells work and sur-
vive, Yang said. Temperature increases could affect how DNA works, for instance,
and change how protein molecular machines operate. Yang also hypothesized that
the cells may use differences in temperature as a means of communication. The team
is working to determine how cellular temperature is regulated, with the goal of ap-
plying the information to improving prevention, diagnosis and treatment of diseases.
We hope that this experiment will change the view of intracellular thermodynam-
ics, encouraging researchers to ask questions about temperature gradient and its pos-
sible roles in signaling, Yang said.
The research was presented Aug. 28 at the American Chemical Societys annual
meeting in Denver.
PASADENA, Calif. High-resolution 3-D
imaging of a cells nucleus undergoing
cell division is now possible, thanks to a
combination of plunge-freezing and a new
method of sample slicing. The findings
open up a biological mystery because they
indicate that some cells take one of the
characteristic steps of mitosis significantly
differently from others.
Traditionally, two sets of chromosomes
pair up at the center of the cells nucleus
during mitosis. Then hollow rods of pro-
tein microtubules composed of a cellular
structure called the spindle apparatus
grab onto the chromosomes and essen-
tially pull each set away from the center in
opposite directions, ensuring that each cell
receives a full copy of the genetic mate-
rial. Typically, in the cells of fungi, plants
and many animals, one or more micro-
tubules attach to each chromosome before
the spindle separates the sets of chromo-
somes from one another.
However, when researchers at Califor-
nia Institute of Technology observed this
step using their new technique, what they
saw was not the usual cell division. In-
stead, they discovered a cell with fewer
microtubules used than chromosomes.
The group used electron cryotomogra-
phy (ECT) to image biological samples.
Unlike traditional electron microscopy,
ECT involves plunge-freezing samples
so quickly that they become trapped in a
near-native state within a layer of transpar-
ent glasslike ice. High-resolution images
can then be captured as the sample is ro-
tated, usually one degree at a time. Beam-
penetration issues have limited ECT to
samples less than 500 nm thick, such as
small bacteria and viruses.
Observing eukaryotic cells
Now, however, the Caltech group has
extended the technique to observe eukary-
otic cells, which typically are much big-
ger. The investigators located the smallest
known eukaryote, Ostreococcus tauri, a
cell with 20 chromosomes, and imaged it
with ECT. Next, they set out to observe
the eukaryotes cell division, but even its
tiny size exceeded the 500-nm limit when
it underwent mitosis.
They used a diamond knife to
cryosect cut a frozen sample into
slices enabling them to look at them
through dividing cells in a near-native hy-
drated state. They made detailed observa-
tions of mitosis in O. tauri.
Contrary to expectations, nowhere near
40 microtubules attached to the two sets of
chromosomes during mitosis. Instead, they
discovered only about 10 small, incom-
plete microtubles, suggesting that the
chromosomes may link together to form a
bundle that can then be segregated all at
once by a smaller number of microtubules.
The findings appeared in the Sept. 8
issue of Current Biology (doi: 10.1016/
j.cub.2011.08.021).
The researchers are now using the same
method to try to image human cells.
b
BIOSCAN
Mitosis: New techniques reveal cell division surprise
EAST LANSING, Mich. An inexpensive
handheld cancer diagnostic device may
soon help physicians in developing nations
with limited resources to detect and diag-
nose the disease and to provide treatment
before its too late.
Cancer is emerging as a leading cause
of death in underdeveloped and develop-
ing countries, and screening for it is al-
most nonexistent because current diagnos-
tics and rapid screening methods are not
suitable for low-income and resource-
limited countries.
Syed Hashsham, a professor of civil and
environmental engineering at Michigan
State University, concentrated his efforts
on developing a low-cost, mobile device.
The instrument, called the Gene-Z, oper-
ates using an iPod Touch or Android-based
tablet and analyzes microRNAs and other
genetic markers. MicroRNAs are single-
stranded molecules that regulate genes;
changes in certain microRNAs have been
linked to cancer and other diseases.
The Gene-Z is battery-operated and
solar-chargeable, making it a suitable fit
for areas where there are limited resources
and mobility is important. The device
can screen for established markers of
cancer at extremely low cost in the field,
Hashsham said.
Until recently, little effort had been
placed on cancer detection for developing
countries. Early detection could lead to
more affordable management of cancers
with the aid of new screening and diag-
nostic technologies that could overcome
global health care disparities, said Reza
Nassiri, director of MSUs Institute of
International Health.
Hashsham, who showcased the test
with colleagues at the National Institutes
of Healths first Cancer Detection and
Diagnostics Conference in August, is
working with Nassiri on the devices
medical capabilities and is establishing
connections with physicians worldwide.
b
BIOSCAN
The handheld Gene-Z, which is operated using
an Android-based tablet or iPod Touch, performs
genetic analysis on microRNAs and other genetic
markers to detect and diagnose cancer.
Handheld units detect cancer on the go
SAN DIEGO Scientists have long
known that plants and animals are gov-
erned by circadian rhythm a 24-hour
cycle of alternating light and darkness to
guide biological processes. Although we
know that our bodies are entrained, or
synchronized, by light and would drift out
of phase if left in the dark, we have yet to
understand exactly how this process works
at the molecular level.
To find out, biologists and bioengineers
at the University of California developed a
model biological system that is simpler
than that of an organism. Led by biology
professor Jeff Hasty, they created a simple
circadian system using a model consisting
of glowing, blinking E. coli. The system
was detailed in the Sept. 2 issue of Sci-
ence (doi: 10.1126/science.1205369).
Combining techniques from synthetic
biology, microfluidic technology and com-
putational modeling, the researchers built
a microfluidic chip containing chambers
with E. coli. Within each bacterium, the
genetic machinery responsible for the bio-
logical clock oscillations was green fluo-
rescent protein, which caused the bacteria
to periodically fluoresce.
The researchers modified the bacteria
to glow and blink whenever arabinose a
chemical that triggered the oscillatory
clock mechanisms of the bacteria was
flushed through the microfluidic chip. This
enabled them to simulate day and night
cycles over a period of minutes rather than
days to better understand how a popula-
tion of cells synchronizes its biological
clocks.
We studied quantitatively how a mini-
mal genetic oscillator in single cells is
capable of picking up the phase of an os-
cillatory external input and relay it to
drive the expression of a gene, Hasty
said. In contrast, the timekeeping systems
of natural organisms use coordinated
groups of complex cell oscillators for gen-
erating the daily rhythms and processing
the environmental inputs to produce the
multiple phased signals that coordinate es-
sential cellular and organismal processes.
Hasty said a similar microfluidic system
could be constructed, in principle, with
mammalian cells to study how human
cells synchronize with light and darkness.
In future research, they hope to integrate
synthetic clocks with cell processes such
as metabolism and cell division to under-
stand how this is achieved in natural
settings.
Genetic models such as this could be
important because circadian rhythm dis-
ruptions have been linked to medical is-
sues such as sleep disorders and diabetes.
While fluorescence microscopy will
continue to be the central tool for the in-
vestigation of biological clocks, the use of
light to control gene expression in individ-
ual cells (optogenetics) will be, without a
doubt, an important new resource to learn
about the dynamics of synthetic multicel-
lular systems, Hasty said.
b
BIOSCAN
Glowing bacteria shed light on what makes biological clocks tick
BUSINESSSCAN
BETHESDA, Md. The US National
Institutes of Health recently awarded
$143.8 million in grants to speed up the
translation of research projects into im-
proved health, and several of the recog-
nized projects rely on biophotonics tech-
nologies or techniques.
These grants were awarded under three
programs supported by the NIH Common
Fund: the NIH Directors Pioneer, New
Innovator and Transformative Research
Projects Awards. This year, approximately
$10.4 million will go to Pioneer awardees,
$117.5 million to New Innovators and
$15.9 million to Transformative Research
Projects.
These programs reinvigorate the bio-
medical workforce by providing unique
opportunities to conduct research that is
neither incremental nor conventional,
said Dr. James M. Anderson, director of
the Division of Program Coordination,
Planning and Strategic Initiatives, who
guides the Common Funds High-Risk
Research program.
The NIH Directors Award Program so
far has funded 406 high-risk research
awards: 111 Pioneer Awards since 2004,
216 New Innovator Awards since 2007
and 79 Transformative Research Projects
Awards since 2009. These numbers in-
clude this years 13 Pioneer Awards, 49
New Innovator Awards and 17 Transfor-
mative Research Projects Awards.
Pioneer Award
Pioneer Award recipient Jean Bennett
of the University of Pennsylvanias Scheie
Eye Institute of the Perelman School of
Medicine and her research team were
granted $4 million for the next five years
to use gene therapy to treat inherited
forms of blindness, which can be caused
by mutations in any of hundreds of differ-
ent genes.
The researchers plan to resensitize the
blind eye using optogenetic techniques in
which light-sensitive molecules are deliv-
ered to any remaining retinal cells. Pre-
clinical studies in blind animals have
demonstrated that this strategy is effective,
and a new clinical study would test the
safety and efficacy of this approach in
blind patients.
Project results could improve the qual-
ity of life for millions of individuals and
also could pave the way for development
of novel gene therapy approaches to the
treatment of other sensory diseases.
New Innovators
New Innovator awards went to several
photonics-focused researchers. Arjun Raj
of the University of Pennsylvanias School
of Engineering and Applied Science will
receive $1.5 million over five years. His
research involves the development and
application of new microscopic imaging
tools to reveal how the physical organiza-
tion of the genetic code determines the
manner in which the cell reads the code
itself. The development of these methods
will establish a nuclear GPS that should
permit researchers to directly visualize
genetic organization in single cells. Under-
standing this organization will be impor-
tant for discovering how defects in trans-
lating the genetic code contribute to
diseases such as cancer.
Other New Innovators named this year
include Bo Huang of the University of
California, San Francisco, who plans to
use superresolution microscopy to study
macromolecular complex architecture in
situ. Huang and colleagues hope that their
research will provide insights into prob-
lems in structural and neuron cell biology,
and generate tools for other life sciences
fields as well.
Long Cai of California Institute of Tech-
nology is exploring systems biology in sin-
gle cells using superresolution bar coding.
The IR-LAMP project, led by Julie C.
Canman of Columbia University, will use
optogenetic technology to spatially manip-
ulate the function of proteins in vivo.
Hongrui Jiang of the University of Wis-
consin-Madison, another New Innovator,
is developing a contact lens to correct
presbyopia by changing its focal length
to offer far-ranging and close-up vision
in a single lens.
Andrea M. Kasko of the University of
California, Los Angeles, will use photo-
tunable biomaterials to engineer complex
3-D cell microenvironments, and Haining
Zhong of Oregon Health and Science
University will examine the architecture
of brain tissue synapses at nanometer
resolution.
Transformative Research Projects
A $7 million five-year Transformative
Research Project Award was given to a
team of investigators from the Perelman
School of Medicine and from Emory
University and Georgia Institute of Tech-
nology, both in Atlanta. The researchers
include Sunil Singhal, director of the
Thoracic Surgery Research Laboratory at
the University of Pennsylvania. If a tumor
is more visible and easier to distinguish
from surrounding tissues, surgeons are
more likely to be able to remove it com-
pletely. At present, a significant number
of patients who undergo surgery leave the
Photonics-related projects score NIH grants
ND11_BusinessScan_Layout 1 10/27/11 10:37 AM Page 21
operating room without total tumor re-
moval.
To address the problem, the researchers
developed fluorescent nanoparticle probes
that target cancer cells. Their main goals
are to help surgeons distinguish tumor
boundaries, identify diseased lymph nodes
and determine whether a tumor has been
completely removed. The grant-funded
project includes plans for tests of the
nanoparticles in animal models and a clin-
ical trial for patients with lung cancer. The
proposed technologies could be broadly
applicable to many types of solid tumors.
Other Transformative Research Projects
awardees include Adela Ben-Yakar and
Jonathan T. Pierce-Shimomura of the Uni-
versity of Texas at Austin, who will use
high-speed optofluidics to study the entire
nervous system as it relates to aging and
disease.
The awards are intended to catalyze
giant leaps forward for any area of bio-
medical research, allowing investigators to
go in entirely new directions, Anderson
said.
Compiled by BioPhotonics staff
b
BUSINESSSCAN
BUSINESSBRIEFS
Avantes BV, a manufacturer of instruments,
light sources, software and accessories for spec-
troscopic applications, has moved to a state-of-
the-art 2700-sq-m facility in Apeldoorn, Nether-
lands. The energy-efficient facility will provide
increased production capacity, enabling shorter
lead times. It also will provide for extended en-
gineering operations to support planned R&D
projects. Avantes released nine new spectrome-
ters in 2011 and plans to release several new
instruments and to announce improvements to
its existing product lines.
Synoptics has established the Advanced Tech-
nology Group, a new division within the com-
pany that will work with life sciences companies
and academic clients to deliver custom imaging
equipment to improve bench-based research,
quality control and clinical development
processes. Comprising the Syngene, Synbiosis
and Syncroscopy divisions, Synoptics provides
products for life scientists to enhance the quality
and speed of their research. Part of the Scien-
tific Digital Imaging Group of companies
based in Cambridge, UK, Synoptics develops
and manufactures digital imaging systems, and
systems and software to improve biological
quality control and research processes.
In Oregon, a collaboration between industry
and academia has resulted in the creation of
a laboratory that will provide researchers with
several state-of-the-art electron microscopes
to advance the understanding and treatment
of cancer, AIDS and other diseases. The OHSU/
FEI Living Lab for Cell Biology, a joint effort
of Oregon Health & Science University
(OHSU) of Portland, where the lab is based,
and FEI Co. of Hillsboro, will be equipped with
a variety of instruments made by the latter.
Among them are a Titan Krios transmission
electron microscope and a Helios NanoLab
DualBeam instrument, which features both
scanning electron and focused ion beam
microscopy functions.
Photonic Solutions of Edinburgh, UK, an-
nounced that, with the addition of the Pharos
and Orpheus ultrafast laser systems, it has
expanded its range of Lithuania-based Light
Conversions products distributed in the UK
and Ireland. The new agreement strengthens an
existing long-term relationship based on sales
of Light Conversions original ultrafast products.
STATEMENT OF OWNERSHIP
MANAGEMENT AND CIRCULATION
of
BioPhotonics
September 30, 2011
(This statement is published in compliance with
the Act of October 23, 1962.)
Published monthly.
Publisher: Karen A. Newman
Editor: Melinda A. Rose
Managing Editor: Laura S. Marshall
The owners are Thomas F. Laurin and Ralph Cianflone,
Berk shire Common, PO Box 4949, Pittsfield, MA 01202-4949.
Known bondholders, mortgagees and other security holders
owning or holding 1 percent or more of total amount of bonds,
mortgages or other securities: None.
The average number of copies of the publication during the 12
months preceding the date shown above is: Printed 23,915;
paid and/or requested subscription by mail 22,186; total paid
and/or requested circulation 22,186; free distribution by mail,
carrier or other means, samples, complimentary and other free
copies 419; total nonrequested distribution 507; office use, left
over, unaccounted, spoiled after printing 1,222; total above
23,915. Actual number of copies of single issue published nearest
to filing date: Printed 21,194; paid and/or requested subscription
by mail 19,997; total paid and/or requested circulation 19,997;
free distribution by mail, carrier or other means, samples, compli-
mentary and other free copies 350; total nonrequested distri-
bution 385; office use, left over, unaccounted, spoiled after
printing 812; total above 21,194.
I certify that the above statements made by me are correct
and complete.
Thomas F. Laurin
President
Laser diode manufacturer and distributor
Frankfurt Laser Co. of Friedrichsdorf, Ger-
many, has announced that it will distribute
Focuslight Technologies Co. Ltd.s high-
power laser diodes. Focuslight, of Xian, China,
provides lasers for applications including pump-
ing, materials processing, display and projection
technologies, printing and the medical field.
The diodes are available from 635 to 1550 nm,
and in both continuous- and pulsed-mode
operation.
Precision positioning systems manufacturer
Physik Instrumente (PI) LP of Karlsruhe, Ger-
many, announced its expansion into the Asian
market with the opening of a direct office in Sin-
gapore. Before opening the office in June, PI was
active in the region through a distributor. PI Sin-
gapore LLP will provide more dedicated service
to its customers, allowing it to develop new busi-
ness in Singapore and greater Southeast Asia.
The Chicago-based John D. and Catherine
T. MacArthur Foundation recently announced
22 new MacArthur fellows for 2011. Recipients
of the Genius Awards receive $500,000 in
no-strings-attached support over the next five
years. They are selected for creativity, originality
and potential to make important contributions
in the future. William Seeley, an associate pro-
fessor of neurology at the Memory & Aging
Center at the University of California, San Fran-
cisco, received an award for his contributions
to neurology. He integrates microscopy, MRI
and clinical examination to explore the struc-
tural, functional and behavioral aspects of
human neurodegenerative disease, concentrat-
ing on frontotemporal dementia.
GE Healthcare of Chalfont St. Giles, UK, is
set to dedicate $1 billion of its total R&D budget
over the next five years to expanding its ad-
vanced cancer diagnostic and molecular imag-
ing capabilities as well as its technologies for
biopharmaceuticals manufacturing and for
cancer research. Announced alongside a $100
million open innovation challenge in New York,
the billion-dollar investment crosses all lines
of GE Healthcares global business and will
enable the company to bring the most promis-
ing cancer ideas to market. The companys
solutions combine medical imaging, molecular
diagnostics and health care information tech-
nology.
Mad City Labs Inc. of Madison, Wis., has
opened a direct sales office in Zurich. The com-
pany said it established Mad City Labs GmbH
to enhance service and support for its European
customers and to strengthen its relationships
with its distribution network and business part-
ners. The Wisconsin company manufactures
piezoactuated, closed-loop nanopositioning
systems for metrology, photonics and micros-
copy applications.
Cosmetic laser maker Palomar Medical Tech-
nologies Inc. of Burlington, Mass., has an-
nounced the receipt of $31 million plus royalties
from aesthetic device company Syneron Med-
ical Ltd. of Yokneam, Israel, in a settlement of
their patent infringement dispute over hair re-
moval systems. The agreement includes two
nonexclusive patent license deals. Under the
first, Palomar granted Syneron and its Candela
unit a worldwide, irrevocable license to US
Patent Nos. 5,735,844 and 5,595,568 for pro-
fessional laser- and lamp-based hair removal
technology. Under the second, Palomar granted
Syneron and affiliates a nonexclusive license in
the US to the same two patents for consumer
home use lamp-based hair removal products.
A supplier of electron microscopes and scientific
instrumentation, JEOL USA of Peabody, Mass.,
and its parent company JEOL Ltd. of Akishima,
Japan, have opened JEOL Brasil Instrumentos
Cientificos Ltda. in Sao Paulo to support the
growing installed base there, and they have
relocated the personnel to a new facility. The
company has a 40-year history in Brazil, much
of it through its agent, Fugiwara Enterprises
IC Ltda. As the number of customers has con-
tinued to grow in the region, the company
decided to provide direct support through its
own service engineers, and administrative and
sales personnel.
3LFR4XDQWIRU6SHFWURVFRS\
3LFR4XDQWIRU6SHFWURVFRS\
)OXRUHVFHQFH/LIHWLPH6SHFWURPHWHU
Time-resoIved uorescence spectroscopy Anisotropy
Decay anaIysis UItra sensitive anaIysis Photochemistry SoIar
ceII research SingIe oxygen MateriaIs research ...
Automated system optimization for each sampIe
SingIe photon sensitivity
Intuitive and user friendIy
Picosecond to miIIisecond timing
Steady-state option
/HDGLQJLQ6LQJOH3KRWRQ&
RXQWLQ
J
$
S
S
OLF
D
WLR
Q
V
PicoQuant GmbH
Berlin, Germany
Tel.: +49-(0)30-6392-6929
info@picoquant.com
www.picoquant.com
Meet us at
ASCB 51st Annual Meeting
December 3-7, 2011
Denver, CO, USA
booth #205
1
(
:
FIuoTime 300
BUSINESSSCAN
b
M
ultimodal nonlinear optical
(NLO) imaging is an emerging
microscopy approach gaining
widespread use in a variety of biomedical
applications. It harnesses and integrates
the unique capabilities of nonlinear
processes such as multiphoton fluores-
cence, second- and third-harmonic genera-
tion (SHG and THG), and coherent Raman
scattering (CRS) and combines them
seamlessly into a single, unified micros-
copy platform. A combination of label-
based and label-free imaging modalities
enables simultaneous acquisition of com-
plementary structural information within
individual cells and elucidates the health
of biological tissue at the submicron level.
The emergence of multimodal NLO
imaging has been facilitated by advances
in ultrafast laser technology; high-perfor-
mance specialized optical filters; and
high-sensitivity detectors.
The development of in vivo diagnostic
tools and techniques that can be used to
advance our understanding of the molecu-
lar, morphological and functional changes
that occur during disease progression is
critical for human health. Today, compre-
hensive interrogation of the structure and
function of biological samples exploits ad-
vances in various optical microscopy tech-
niques and modalities. Wide-field and
laser-scanning confocal fluorescence mi-
croscopy have been used extensively to
elucidate the form and function of biologi-
cal entities at the submicron level. More
recently, however, advances in ultrafast
laser technologies have allowed the ex-
ploitation of several well-known nonlinear
optical processes for use in biomedical
imaging applications.
Two common, well-known nonlinear
imaging techniques are multiphoton fluo-
rescence
1
and coherent Raman scattering
2
microscopy. The former typically uses
femtosecond near-infrared laser pulses,
while the latter, based on either coherent
anti-Stokes Raman scattering (CARS) or
stimulated Raman scattering (SRS), uses
two separate picosecond near-infrared
lasers to generate a CRS signal. As a re-
sult, it is not natural or obvious to com-
bine these two imaging techniques into a
single system.
Advances in ultrafast laser technology;
specialized optical filters for transmitting,
reflecting and blocking various light sig-
nals; and high-sensitivity detectors now
permit the construction of a single multi-
modal nonlinear optical microscope plat-
form that offers both label-based and
label-free imaging strategies with which
to interrogate biological samples.
Multimodal NLO microscopy combines
several imaging modalities into a single
To Label or Not?
Figure 1: Multimodal nonlinear optical (NLO) imaging of a mouse kidney: (a) CARS, (b) two-photon
autofluorescence and (c) second-harmonic generation. All three signals are displayed in a single,
co-registered image in (d). The series of images of sections of mouse kidney has helped researchers
understand the impact of fat content on cardiovascular disease and diabetic conditions in animal
models. Images courtesy of professor Eric O. Potma, University of California, Irvine.
Just as research-based studies
laid the path for clinical
investigations with
intravascular OCT and
near-infrared
reflectance spectroscopy,
todays pioneering research
likely will result in a future
in which multimodal
nonlinear optical imaging
will serve as a potent weapon
with which to clinically
investigate, understand
and diagnose a variety of
human health factors.
BY DR. NEIL ANDERSON, SEMROCK INC., A UNIT OF IDEX CORP.
ND11_Semrock Feat_Layout 1 10/27/11 10:21 AM Page 24
unified platform and affords a myriad of
benefits. Chief among these are: inherent
3-D sectioning capability; the ability to
image deeper into samples, such as tissue,
using NIR (700- to 1100-nm) light com-
pared with linear microscopy, where visi-
ble (400- to 700-nm) light is used; the use
of lower laser average power to prevent
photodamage and phototoxicity compared
with laser-scanning confocal microscopy;
and the ability to exploit and combine
label and label-free imaging modalities.
In Figure 1, the utility of multimodal
NLO imaging shows representative com-
posite images of a section of kidney tissue
from a mouse model. The image contrast
is provided by collecting the (a) anti-
Stokes Raman light associated with the
CARS process, (b) two-photon autofluo-
rescence emission and (c) the SHG signal.
A composite image of all three signals is
shown in (d) and demonstrates how each
co-registered signal can be combined to
provide a comprehensive portrait of the
sample.
The most common modalities used in
multimodal NLO imaging are multiphoton
fluorescence, SHG and THG, and CRS.
Each provides unique and complementary
benefits. Two- and three-photon fluores-
cence microscopy exploit the intrinsic
two- and three-photon absorption-induced
excitation of various organic dyes, fluores-
cent proteins and quantum dots that are
typically used to label samples fluores-
cently. Long-wavelength ultrafast laser
pulses have sufficient peak intensity to
enable a high probability that two or three
photons arrive at the molecule simulta-
neously, raising the fluorescent molecule
into an excited state. The fluorophore then
relaxes back to the ground state, produc-
ing fluorescence emission.
A key limitation of this approach is that
it requires the addition of exogenous fluo-
rophores. Label-free two-photon imaging
is possible by selectively targeting intrin-
sic cellular molecules, such as NADH
and flavins.
3
This approach is commonly
called two-photon autofluorescence imag-
ing. Label-free multiphoton imaging also
is possible via three-photon processes and
can be used to image tryptophan and sero-
tonin, for example.
3
Alternative label-free approaches to cel-
lular-level imaging exploit both SHG and
THG nonlinear processes within the sam-
ple. Collagen proteins exhibit noncentro -
symmetric molecular organization and
thus can be imaged by collecting the emit-
ted SHG signal under laser excitation.
4
THG occurs even in centrosymmetric
structures and is useful for imaging inter-
face heterogeneities as well as myelin
sheaths in studies of the nervous system in
animal models.
5
Another benefit of both
SHG and THG imaging is that, because
neither process involves the coupling of
electronic levels, photobleaching effects
can be suppressed, enabling structures to
be observed over extended periods.
Chemically specific imaging also is
possible and can be achieved by incorpo-
rating CRS into the same microscope plat-
form. CRS microscopy comes in two
unique forms: CARS and SRS.
6
Both offer
Raman imaging with a better signal-to-
noise ratio and higher sensitivity than
spontaneous Raman microscopy. CARS
microscopy is the most commonly imple-
mented of the two in multimodal NLO
imaging, but SRS is rapidly growing in
popularity.
A typical multimodal NLO imaging sys-
tem consists of a laser-scanning confocal
microscope platform equipped with two
ultrafast near-infrared laser sources; vari-
ous interference filters for transmitting,
reflecting and blocking light signals; and
multiple high-sensitivity detectors to col-
lect each of the independent light signals.
One possible configuration for a multi-
modal NLO microscope for simultaneous
two-photon fluorescence, SHG and CARS
microscopy is shown in Figure 3.
In this example, two-photon fluores-
cence and SHG imaging can be performed
using a single widely tunable near-infrared
femtosecond or picosecond laser. Intro-
ducing a second picosecond laser com-
monly referred to as the Stokes beam (at
the optical frequency
Stokes
) enables
CARS microscopy. When this beam is
spatially and temporally overlapped with
the beam of the first laser commonly re-
ferred to as the pump laser (at frequency
pump
) the beams interact within the sam-
ple via a four-wave mixing process to gen-
erate the CARS signal at the new optical
frequency
CARS
= 2
pump

Stokes
. Both
the Stokes and pump beams are collinearly
Figure 2: Energy level diagrams that describe the nonlinear processes exploited in multimodal nonlinear optical imaging: (left to right) third-harmonic
generation (THG), second-harmonic generation (SHG), two-photon (2P) and coherent anti-Stokes Raman scattering (CARS). In the THG process, three photons
at wavelength are required to generate the THG signal at wavelength /3. In the SHG process, two input photons () are required to generate the
SHG signal (at /2). In two-photon fluorescence, an excitation source at wavelength with sufficient peak intensity is used, such that there is a high
probability of absorbing two photons simultaneously, thus producing fluorescence at a wavelength just longer than the /2. The CARS process
is a nonlinear four-wave mixing process that involves three input photons to generate the CARS signal at
CARS
= 2
pump

Stokes
.
overlapped and directed to a microscope
head via a specially designed notch
dichroic beamsplitter positioned at 45.
Through judicious wavelength selec-
tion, direct coupling of the characteristic
vibrational modes that uniquely identify
structures of interest within the sample can
be achieved. In CARS imaging, the Stokes
beam (
Stokes
) is typically fixed at a wave-
length of 1064 nm, while the pump beam
(
pump
) is typically produced by an optical
parametric oscillator (OPO), which can be
tuned across the 700- to 900-nm-wave-
length range. When the pump beam is
tuned to a wavelength of approximately
816 nm, the frequency difference (
CARS
)
is ~2850 cm
1
, which corresponds to the
symmetric CH
2
stretching vibration in
lipids.
In this example, the same pump beam
generated by the OPO laser at approxi-
mately 816 nm also can be used to gener-
ate the two-photon fluorescence and SHG
signals within the sample. To avoid inter-
ference from the CARS signal, the Stokes
beam can be blocked. Alternatively, to
permit simultaneous multiphoton fluores-
cence and CARS imaging, the Stokes
beam is allowed to pass to the microscope.
Both the CARS and SHG signals are de-
tected in the forward-scattering geometry,
while the two-photon fluorescence emis-
sion is detected in the backward (epi)
geometry.
Just as important as the right laser
sources is the choice of appropriate optical
filters matched to sensitive detectors. For
high-contrast multimodal NLO imaging, it
is critical that the proper filters are used to
transmit and reflect the input laser beams
and to transmit the multiphoton and CARS
signals. Characteristics that require careful
consideration are: greater than 95 percent
average transmission across the passband;
steep edges; and high out-of-band block-
ing. Controlling and managing the level of
undesired out-of-band light, such as the
bleedthrough from the source lasers, is
critical. Any stray laser light that reaches
the detectors can significantly affect the
acquisition of high-fidelity i.e., high-
Multimodal Imaging
Figure 3: In this schematic of a multimodal nonlinear optical microscope, two ultrafast laser
sources operating in the near-infrared are used to generate two-photon fluorescence emission,
second-harmonic generation and coherent anti-Stokes Raman scattering signals within a sample.
Dichroic filters are used to transmit and reflect the laser light into the microscope and to direct the
desired signals to each detection channel. Each photomultiplier tube detector is matched with an
appropriate emission filter to transmit the desired signal and block all out-of-band light.
Figure 4: Examples of filter options for multimodal nonlinear imaging: dichroic/emission pair
for multiphoton fluorescence, SHG and THG microscopy (left), and CARS microscopy (right).
signal-to-noise-ratio images. Therefore,
dichroic filters that transmit and reflect
only the desired light signals as well as
emission filters that provide high blocking
(optical density >6 to 8) over an extended
wavelength range are required. Examples
of filter options for multimodal nonlinear
imaging are shown in Figure 4.
Photomultiplier tubes (PMTs) are the
most common detectors used in multi-
modal NLO imaging. PMT detectors
based on gallium arsenic phosphide are a
suitable choice for detecting light across
the visible range (400 to 700 nm). Alterna-
tively, red-sensitive PMT detectors, based
on multialkali metals e.g., Sb, Na, K,
Cs that provide high sensitivity across
the visible and extend out into the near-
infrared (>900 nm) also are popular
choices. Continued improvements in
detector sensitivity, reduction in footprint
and the advent of a less complex, easy-
to-use modular design have all benefited
NLO imaging applications.
Optical microscopy continues to play
an increasingly important role in biomed-
ical research. To date, techniques such as
wide-field, confocal and multiphoton
microscopy have provided a wealth of
insight and information on the form and
function of a myriad of samples, such as
live cells and biological tissue. By har-
nessing nonlinear imaging modalities into
one unified platform, multimodal NLO
imaging provides an even more powerful
microscopy-based approach with which to
study the health and function of biological
specimens.
One benefit of this approach in medical
research has been the use of multimodal
NLO imaging to advance our understand-
ing of cardiovascular disease, a leading
cause of illness and death globally. To elu-
cidate the impact of diet, researchers at the
University of California, Irvine, used mul-
timodal NLO imaging to understand how
dietary fat content affected kidney health
in animal models. They varied the fat con-
tent in the diet fed to animal subjects over
a period of weeks. Post-study analysis
using CARS, two-photon and SHG imag-
ing of the kidney allowed them to quanti-
tate the levels of accumulated fat deposits
within the kidney and their impact on sur-
rounding structural components, such as
collagen fibers. Multimodal NLO images
from this study are shown in Figure 1.
The researchers found a correlation
among all three nonlinear co-registered
signals, as shown by the composite image
(d). CARS imaging clearly revealed the
accumulation of lipid deposits within the
kidney and was used as a quantitative
measure for the fat content. They used the
collected two-photon autofluorescence sig-
nal to locate and identify important struc-
tures such as individual cells within the
kidney tubules and the SHG imaging not
only to identify collagen content within
the mouse kidney, but also to understand
the impact of fat content on the level of
collagen present as well as on collagen
fibrillogenesis.
Multimodal NLO imaging will continue
to find widespread use and growing popu-
larity in the life and health sciences. Con-
tinuing developments in ultrafast laser
sources, specialized hard-coated optical
interference filters and high-sensitivity
detectors make it straightforward to com-
bine various nonlinear imaging modalities,
such as CARS, two-photon, SHG and
even THG into a single, unified micro-
scope platform.
Meet the author
Dr. Neil Anderson is a technology development
analyst with Semrock Inc., a unit of Idex Corp.,
in Rochester, N.Y.; e-mail: nanderson@idex
corp.com.
Acknowledgment
The author thanks professor Eric O.
Potma, University of California, Irvine,
for supplying the data used in Figure 1.
References
1. W.R. Zipfel et al (2003). Nonlinear magic:
multiphoton microscopy in the biosciences.
Nat Biotech, Vol. 21, Issue 11, pp. 1369-
1377.
2. C.L. Evans and X.S. Xie (July 2008). Coher-
ent anti-Stokes Raman scattering micros-
copy: chemical imaging for biology and
medicine. Annual Rev of Analyt Chem,
Vol. 1, No. 1, pp. 883-909.
3. W.R. Zipfel et al (2003). Live tissue intrinsic
emission microscopy using multiphoton
excited native fluorescence and second-
harmonic generation. PNAS, Vol. 100,
No. 12, pp. 7075-7080.
4. C.P. Pfeffer et al (October 2008). Multimodal
nonlinear optical imaging of collagen arrays.
J Struct Biol, Vol. 164, Issue 1, pp. 140-145.
5. M.J. Farrar et al (2011). In vivo imaging of
myelin in the vertebrate central nervous
system using third harmonic generation
microscopy. Biophys J, Vol. 100, Issue 8,
pp. 1362-1371.
6. C.W. Freudiger et al (2008). Label-free bio-
medical imaging with high sensitivity by
stimulated Raman scattering microscopy.
Science, Vol. 322, pp. 1857-1861.
Americas +1 505 296 9541
Europe +31 (0)316 333041
Asia +81 3 3407 3614
For more information:
www.cvimellesgriot.com/Ultrafast
Low dispersion
Broad bandwidLh
Hiqh damaqe Lhreshold
CaLaloq and cusLom opLions
Ultrafast Optics
When every pulse counts
Wavelength (nm)
730 750 770 790 810 830 850 870
-50
-40
-30
-20
-10
0
10
20
30
40
50
R
e
f
l
e
c
t
a
n
c
e

G
r
o
u
p
D
e
l
a
y

D
i
s
p
e
r
s
i
o
n

(
f
s
2
)
traditional broadband
traditional high LDT
Ti: Sapphire Mirror
Mirrors, beamsplitters, polarizers and
prisms for Ti:Sapphire or ber lasers
applications.
Multimodal Imaging
Retina Div., at Johns Hopkins Hospital in
Baltimore. Typically, these procedures
are performed in the outpatient clinic with
the use of topical anesthetic eyedrops.
A contact lens is positioned onto the eye
to lubricate the cornea and hold open the
eyelid. The laser settings including
power, spot size, pulse duration and repeti-
tion rate are selected, and the aiming
beam is focused through the pupil onto
the desired areas for retinal treatment.
Recent studies have demonstrated that
the science behind photocoagulation is
more complicated than once thought and
that heating produces a constellation of
changes at the molecular level in individ-
ual cells; e.g., photocoagulation creates a
surge in free radicals.
Progress so far
Developments in laser technology have
made lasers of various wavelengths possi-
ble, and the wavelength selected depends
upon the type of procedure being per-
formed.
The wavelengths typically used for reti-
Laser photocoagulation can be used
to treat the wet form of AMD, in which
blood vessels start to leak into the retina,
creating fogging and, later, a blind spot in
the center viewpoint. Other ocular condi-
tions that can be treated include prolifera-
tive diabetic retinopathy, diabetic macular
edema, vascular occlusions, central serous
chorioretinopathy, retinal tears, tumors,
choroidal neovascularization, arterial
macroaneurysms and retinopathy of
prematurity.
The treatment strategy utilized for pho-
tocoagulation varies, depending upon the
particular disease process, said Dr. Adri-
enne W. Scott, assistant professor of oph-
thalmology at The Wilmer Eye Institute,
S
ocrates first described nonlaser
photocoagulation in 400 B.C. after
observing solar eclipse burns of the
retina. But it was not until an afternoon
in July 1945 when an unfortunate medical
student viewed a partial eclipse of the sun
in Northern Europe that the potential
benefits of light burns to the eye were
first realized.
The student in question worked under
German ophthalmologist Dr. Gerhard
Meyer-Schwickerath, who came to the
conclusion in 1947 that an evolving retinal
detachment stops at a scar. The idea to
produce artificial scars by intense light
illumination came to him in the same year
during a restless night where he wrote two
words, light and coagulation, on a
sheet of paper, so as not to forget the
thought by morning.
In 1957, the light coagulator was pre-
sented by Carl Zeiss as the first commer-
cially available photocoagulation device,
and more than 1000 were sold until the
late 1970s.
But it was the invention of the laser in
1960 that catapulted this technology into
ophthalmology. Since that time, photoco-
agulation lasers have been reduced contin-
uously in volume, weight and price.
Although photocoagulation is used in
almost every surgical specialty e.g.,
lasers to treat vascular lesions, benign and
malignant tumors, and bleeding ulcers
Dr. Matthias Schulze, director of market-
ing at Coherent Inc., believes that the use
of photocoagulation in ophthalmology is
the most important because of the growing
market: More and more older people are
suffering from vision deterioration such as
that resulting from age-related macular de-
generation (AMD), the leading cause of
blindness in the Western world.
Photocoagulation the deliberate destruction of tissue by absorption of
radiant energy and its conversion to heat is a treatment undertaken to
save another area of tissue deemed more valuable. It has become a
primary tool in the treatment of myriad ocular conditions.
BY MARIE FREEBODY
CONTRIBUTING EDITOR
Healing Through Laser Damage
Photocoagulation:
A fundus photograph showing treatment with single-spot conventional photocoagulation burns (bottom left)
and adjacent areas treated with patterned scanning laser photocoagulation (top right). Courtesy of Dr. Yannis
M. Paulus.
nd11_FeatPhotocoagulation_Layout 1 10/27/11 10:23 AM Page 28
said. Yellow light is an absorption maxi-
mum of oxyhemoglobin, allowing for
better selective treatment of vasculature
and more uniform absorption.
numerous reasons. It is less affected by
small-angle scattering in the transparent
ocular media and provides greater trans-
mittance through corneal opacities, he
nal photocoagulation range from ~400 to
800 nm. This spans the majority of the
visible electromagnetic spectrum (violet,
380 nm, to red, 750 nm) and part of the
infrared spectrum (750 nm to 1 mm).
The ideal wavelength is characterized
by good penetration through ocular media
and maximal absorption in the target tis-
sue. Shorter wavelengths are more easily
scattered; hence, red light (620 to 750 nm)
has better penetration than blue light (450
to 495 nm), Scott said. Scatter is a result
of radiation absorption by tissues other
than the target. It can occur anywhere
anterior to the retina, including the
anterior segment, lens and vitreous.
The degree of scattering increases with
the maturity of the cataract a clouding of
the lens of the eye and vitreous hemor-
rhage. In such cases, Scott suggests that a
longer wavelength, increased laser duration
or higher energy levels may be required.
Typical laser power levels range from
1.5 to 8 W. The power often is adjusted
during a procedure to ensure that the de-
sired burn is applied over the surface of
the eye. This approach is known as titra-
tion and is necessary because laser power
can vary dramatically among individuals.
Table 1 summarizes the systems, wave-
lengths and pulse durations of currently
available laser photocoagulators.
The green laser wavelength is the most
commonly used laser for photocoagulation
and is mainly employed in two systems:
argon gas (514.5 nm) and solid-state
frequency-doubled Nd-YAG (532 nm),
Scott said. Argon green laser confers the
advantage of a strong affinity for melanin
and hemoglobin with minimal absorption
of xanthophyll, the predominant pigment
in the macula.
However, Dr. Yannis M. Paulus at
Stanford Universitys Byers Eye Institute
at Stanford in California explains that the
yellow laser (577 nm) is preferable to
green in macular and vascular diseases for
Laser Type
CO
2
(carbon dioxide) 9.2 to 10.8 m CW or ms-pulsed
OPSL (optically pumped 480 to 580 nm CW or ms-pulsed
semiconductor laser)
Argon 488, 514 nm CW or ms-pulsed
HeNe 533 nm CW
Nd:YAG 1064 nm CW 100 ns
Nd:YLF 1053 nm CW 10 ps
Diode 635 to 1550 nm CW 2 ns
Dye 450 to 900 nm CW 100 fs
Ruby 694 nm 1 to 250 s
Krypton 531, 568, 647 nm CW or ms-pulsed
Ti:sapphire 700 to 1000 nm 60 fs to 10 ps
Alexandrite 720 to 800 nm 100 s to 50 ns
Er:YAG 2.94 m 100 ns to 250 s
Er:YSGG 2.78 m 100 ns to 250 s
Ho:YAG 2.12 m 100 ns to 250 s
Wavelength Pulse Duration
Coherents Genesis 577 laser is used in photocoag-
ulation applications. Courtesy of Coherent Inc.
Types of photocoagulation lasers (CW = continuous wave). Courtesy of Dr. Yannis M. Paulus, Byers Eye
Institute at Stanford, Stanford University.
Histology of light retinal photocoagulation lesions produced with 7-ms-pulse-duration burns reveals healing of
photocoagulation lens over four months, resulting in almost continuous normal retinal morphology. Reprinted
with permission from the Association for Research in Vision and Ophthalmology, from Y.M. Paulus et al (2008).
Healing of retinal photocoagulation lesions. Invest Ophthal Vis Sci, Vol. 49, Issue 12, pp. 5540-5545.
Until recently, yellow lasers were lim-
ited to costly tunable dye and Krypton
lasers. However, this changed in 2008
with the advent of optically pumped semi-
conductor lasers based on near-infrared-
pumped quantum-well structures.
For Coherents Schulze, the most versa-
tile systems feature green, yellow and red
in a combined tool.
Better understanding, better treatment
Ophthalmologists and laser makers
alike are striving toward a greater under-
standing of the mechanism of action in
photocoagulation so that minimally trau-
matic retinal lasers can be designed for
safe and effective use in any area of the
retina, including the macula.
We have come to understand that the
therapeutic effect from photocoagulation
is not only from tissue destruction but
rather from a complex biological response
to photocoagulation, Paulus said. If we
can induce this therapeutic effect without
causing permanent retinal scarring, we
could treat the entire retina in a short
period of time without worrying about
protecting vital structures, and even
retreat areas of retina.
Rather than watching for a visible
change in the retina, one aim is for real-
time monitoring of retinal temperature
during laser coagulation.
For many coagulator manufacturers,
one of the open issues in photocoagulation
is choosing the right dosage of laser en-
ergy during a treatment session.
The retina of an elderly patient suffer-
ing from severe diabetic retinopathy is
strongly varying in pigmentation, absorp-
tion and scattering losses from point to
point; therefore, for a proper therapy re-
sult, the laser power should be adjusted
individually for every laser spot.
In daily clinical practice, this is impos-
sible when setting up to 2500 lesions
within a single photocoagulation session.
The titration process typically involves
starting with a single spot (or a small pat-
tern) in the midperiphery and increasing
the laser power until a proper whitening
of the retinal tissue occurs.
After this initial setting, the laser energy
is changed if the whitening disappears or
is becoming far too strong. This typically
occurs five to 10 times a session. The re-
sult is areas of over- and undertreatment,
which becomes worse in multispot therapy.
Some medical photonics companies
are working on an individual, feedback-
controlled laser treatment to optimize
outcomes and minimize side effects.
Low-dose laser technologies might have
a certain effect on one point of the retina
but a zero effect if the tissue slightly
alternates.
Although not yet clear whether supra -
threshold, just-above-threshold or sub-
threshold laser therapy would provide the
maximum benefit for the patient, there is
confidence that physicians will figure this
out by using automated dosimetry-con-
trolled photocoagulation therapy lasers.
Once this clinical work is completed, we
should know which treatment option will
be best and whether image-guided laser
therapy is clinically needed or only a nice-
to-have but expensive feature.
marie.freebody@photonics.com
Photocoagulation
The Navilas laser platform showing preplanning
of laser spots. Courtesy of OD-OS Retina Naviga-
tion Co.
This fundus photograph shows laser burns after application with the OptiMedica Corp.-designed pattern scan
laser photocoagulator PASCAL (now produced by Topcon Corp. of Tokyo). The circular and square patterns
were made in <1 s with a scanning laser and shorter pulse duration. Courtesy of Dr. Yannis M. Paulus.
A wide-field fluorescein angiogram of a patient with
diabetic retinopathy demonstrates profound periph-
eral capillary dropout. The white arrow points out
areas of the peripheral retina that appear dark on
fluorescein angiography because of the absence of
the normal retinal capillaries in the setting of diabetic
retinal ischemia. Courtesy of Dr. Adrienne W. Scott.
This is the same patient, one week after scatter laser
photocoagulation to the peripheral retina using a
targeted argon green laser (PASCAL) on the areas
of poor retinal circulation. The laser lesions, indi-
cated by the white arrow, are seen as round dark
spots within the peripheral retina. Courtesy of
Dr. Adrienne W. Scott.
B
iomedical researchers most often
carry out rapid assays of large cell
populations on a single-cell basis
with a flow cytometer. The instrument
originated from research using cell count-
ing with laminar flow in the 1950s and
from fluorescence measurement in the
1960s. The device can achieve rapid assay
with a laminar flow to hydrodynamically
focus the core fluid, which contains the
cells.
1,2
The narrowed core fluid then
transports the cells in single file through
one or more interrogating beams of inci-
dent light.
The optical excitation of a flowing
cell leads to two types of light signal
emissions. The first type is called a scat-
tered light signal, which has the same
wavelength as the incident light and arises
from the heterogeneity of the cells refrac-
tive index relative to the surrounding
medium. The second type of emission is
made up of fluorescent signals from wave-
lengths that usually are larger than the
Diffraction-Imaging
Flow Cytometry
Enables Rapid Cell Assay
A new method allows fast,
label-free cell classification by
combining high-contrast image
acquisition with automated
analysis. This method could
open doors for expanded
clinical applications such as
classification of blood and
tumor cells based on their
3-D morphological features.
BY XIN-HUA HU, EAST CAROLINA UNIVERSITY AND WAVMED TECHNOLOGIES CORP.;
YUANMING FENG, TIANJIN UNIVERSITY; AND JUN QING LU, EAST CAROLINA UNIVERSITY
Figure 1. Two views of reconstructed structures of (a) a Jurkat cell derived from a T lymphocyte and (b) an
NALM-6 cell derived from a B lymphocyte. The dark-blue surfaces are cell membranes, the light-blue surfaces
are aggregated mitochondria, and the green are nuclear membranes. (c) Two simulated diffraction images of
side scatters are shown from the same cell structure: The left corresponds to a cell membrane surface with the
same index of refraction as that of cytoplasm at n
m
n
c
1.35; the right corresponds to the same structure,
but its cell membrane surface index is set at n
m
1.50. Courtesy of the authors.
nd11_FeatureFlow_Layout 1 10/27/11 10:24 AM Page 31
wavelength of the incident light; they are
caused by fluorescent molecules inside
the cell. Because most biological cells are
not self-fluorescing, cells often must be
stained with fluorescent reagents before
the flow cytometer can measure them.
Although both types of light signals
are acquired from the excited cells for
analysis and classification, existing flow
cytometers depend mainly on fluorescent
signals because the conventional designs
for these instruments have limited ability
to extract information from scattered light
signals.
Existing flow cytometers can be di-
vided into two categories according to
their approach to signal acquisition. Most
are designed with a single light detector
such as a photomultiplier or photodiode
to measure signals from both scattered
light and fluorescence from stained cells.
3
Since both signals are relatively strong in
relation to background noise, this design
permits signal measurement and acquisi-
tion during a time window as short as
about 10 s; this provides a foundation
for collecting light signals from flowing
cells at rates of up to 10
4
cells per sec-
ond, or millions of cells in less than half
an hour. At that speed for single-cell
analysis, no other instrument even comes
close, which is the main reason for its
increasing popularity among biomedical
researchers.
In 2005, another flow cytometer design
began to emerge. Its approach to image
acquisition is based on early research
dating from the 1980s.
4
This system essen-
tially combines a conventional microscope
with a flow cytometer to acquire nondif-
fraction images of cells very quickly. Most
flow cytometers, though, rely mainly on
fluorescent light, which can provide multi-
ple images per flowing cell, offering addi-
tional information on morphology. This
benefit comes with a cost, however:
lower throughput of 10
3
or fewer cells
per second.
Conventional cytometers: Drawbacks
Despite their wide applications, conven-
tional flow cytometers do have their draw-
backs. Fluorescent signals yield important
molecular signatures for the interro gated
cells, but a single detector can tell only
whether and how many labeling fluo-
rescent molecules exist in a cell. Morphol-
ogy information is acquired, but the
nondiffraction images yield only 2-D
projections of 3-D cell structure. More
importantly, these nondiffraction images
contain complex patterns that are ex-
tremely difficult to interpret using auto-
mated image analysis with current com-
puter algorithms. Considering that the
data stream from an imaging flow cytome-
ter can yield huge amounts of data in a
short amount of time, the lack of auto-
mated image analysis in real time can
present a significant roadblock to taking
full advantage of the images for analysis
and classification of cells.
Second, measuring fluorescent signals
often requires staining cells with multiple
reagents, which is a time-consuming
process and adds extra costs. In fact, it is
widely known that reagent purchase is a
major factor associated with the high cost
of using flow cytometers. Finally, it is al-
ways possible for the fluorescent reagents
to interfere with various biochemical
processes within the cells under study,
and additional tests may be needed to
verify and/or minimize this possibility.
Scattered light signals
It has long been known that diffraction
imaging methods can be used to acquire
3-D structures of an object under coherent
illumination from scattered signals. Soon
after the discovery of x-rays more than a
century ago, Max von Laue and others
found that the patterns of x-ray photons
scattered from a crystal sample as
recorded by a film can be associated with
3-D lattice structure and constants of the
crystal. The observed patterns were used
to illustrate the wave nature of x-ray radia-
tion as a result of interference or diffrac-
tion among the coherently scattered pho-
tons. The results of diffraction research led
directly to the birth of x-ray crystallogra-
phy, which is still widely used by scien-
tists to reconstruct biomolecules in 3-D.
Extending this 3-D image reconstruc-
tion method to the optical domain is much
more difficult, it turns out, because of the
need for highly coherent light sources.
Dennis Gabor was probably the first to
Flow Cytometry
Figure 2. The flow chamber is based on a jet-in-fluid design concept that uses the objective to collect side
scatters for diffraction imaging.
demonstrate experimentally the feasibility
of 3-D reconstruction with optical light.
5
However, his significant discovery had to
wait for the birth of lasers more than a
decade later to be fully appreciated as a
high-quality 3-D imaging modality.
As tools for light scattering models
have become available, researchers have
realized that 3-D reconstruction of an
object is possible without a reference
beam if a sufficient number of diffraction
images are acquired. Nevertheless, 3-D
reconstruction of cell structures for rapid
assay and classification is difficult, if not
impossible, in a flow cytometer setting
because multiple diffraction images are
needed, and time-consuming inverse
calculations must be performed.
Over the past 10 years, we have con-
ducted research on modeling and have
performed experimental studies on light
scattering by biological cells.
6-8
The results
have led to the recent development of a
new diffraction imaging approach using
flow cytometers.
9-11
By using confocal
fluorescence microscopy and staining the
nucleus and mitochondria with various
reagents, we made realistic 3-D recon-
structions of cell structures. The technique
allowed us to import precisely sized struc-
tures into a computer model for accurate
simulations of light scattering by single
cells. Figures 1a and 1b present 3-D cell
structures viewed from two perspectives.
By assigning different refractive index
values to the intracellular components and
solving Maxwells equation, we can obtain
the angular distribution of scattered light
fields from a cell. With this numerical
modeling tool, diffraction images can be
simulated by projecting the scattered light
intensity on a plane representing an imag-
ing sensor.
Using different 3-D distributions of
intracellular refractive index, diffraction
images can be simulated that yield insight
into the correlation between diffraction
image patterns and 3-D morphological and
index distribution features of illuminated
cells. Two examples of simulated images,
shown in Figure 1c, demonstrate the effect
of intracellular distribution of refractive
index. Taken together, these results moti-
vated us to incorporate the diffraction im-
aging into flow cytometry through a new
approach an alternative to reconstruction
in which the image data are analyzed in
real time as the fingerprints of 3-D opti-
cal structures for rapid assay of cells. We
have also performed goniometric measure-
ment of scattered light signals from cell
suspension samples and have found that
polarization-based measurement can pro-
vide additional handles for cell classifica-
tions, as expected from modeling results.
Designing a new cytometer
Armed with these new insights, we set
out to investigate various designs for the
flow nozzle and flow chamber for the
diffraction imaging flow cytometer we
were developing. These elements play a
key role in forming a hydrodynamically
focused laminar flow positioning cells
accurately at the optical focus of the inter-
rogating laser beam without any index-
mismatched interfaces close to the excited
cell and at the desired speed.
Several groups have published various
designs for flow chambers that acquire dif-
fraction images from cells; these include a
glass flow cell with a square, narrow chan-
nel and a waveguide-based beam-in-ow
design.
12,13
None of these fit our require-
Flow Cytometry
Figure 3. Normalized diffraction images acquired from (a, b) Jurkat cells derived from T lymphocytes and (c, d) Ramos cells derived from B lymphocytes using two 12-bit
CCD cameras with an excitation wavelength of 532 nm for each flowing cell: (a) and (c) are images of side scatters with vertical polarization; (b) and (d) are images of
side scatters with horizontal polarization. The polarization direction of the incident light beam is vertical (top row), 45 from vertical (middle row), and horizontal (bottom
row). The three numbers on the right side of each image are the minimum, average and maximum pixel readings of the 12-bit images before normalization.
ments for acquiring high-contrast diffrac-
tion images with ease of alignment.
After numerous tests and design im-
provements, we finally arrived at a jet-in-
fluid flow chamber design to meet the
above requirements. It incorporates a noz-
zle built in-house to guide the sheath and
core fluids into a water-filled cuvette with
its sides several millimeters away from the
core fluid. The two fluids from the nozzle
form a laminar flow in which the core fluid
narrows to an exit tube in the cuvette. A
3- to 5-mm-long gap space exists between
the extending tip of the core channel in the
nozzle and the exit tube to allow efficient
collection of scattered light from the bio-
logical cell by the CCD camera, without
any noise-producing index-mismatched
interfaces within the field of view. With
the incident laser beam focused at the core
fluid just above the exit tube, the position-
ing of the flowing particles at the laser
beams waist can be accurately achieved
in the gap space with no heterogeneity
of refractive index near the excited cell.
Figure 2 exhibits a close-up view of the
nozzle and chamber.
After the initial development stage, we
constructed two experimental prototypes
of a diffraction imaging flow cytometer
for conducting cell measurements at our
labs at East Carolina University and Tian-
jin University. The current design allows
for the acquisition of two polarization-
resolved images per excited cell using a
polarizing beamsplitter behind the infinity-
corrected microscope objective, which
collects side light scatters. Exploiting
polarization information from diffraction
image data can provide additional parame-
ters for cell analysis and classification,
and data from various polarization states
of the incident laser beam can be incor -
porated. By adopting the gray-level co-
occurrence matrix algorithm widely used
for analyzing human fingerprints, we can
extract multiple parameters at a rate of 70
images per second using a 2-GHz laptop
computer for cell classification.
With further optimization of the auto-
mated image analysis code and parallel
execution on multiple GPUs, we expect
that, in the near future, real-time diffrac-
tion image processing for cell classification
can be scaled up to a rate of 10
3
images per
second. Figure 3 shows some examples of
recent diffraction image data collected
from cultured cells derived from B and T
lymphocytes from cancer patients. Quick
visual examination indicates that differ-
ences in pattern and intensity may be used
to perform label-free classifications of
these two lymphocyte-derived cell types.
A detailed study to test this hypothesis
using the gray-level-co-occurrence matrix
algorithm is in progress.
Meet the authors
Dr. Xin-Hua Hu is a physics professor at East
Carolina University in Greenville, N.C., and
chief scientist at WavMed Technologies Corp.
in Tianjin, China; e-mail: hux@ecu.edu.
Dr. Yuanming Feng is a professor and chairman
of the biomedical engineering department at
Tianjin University in China; e-mail: ymfeng
@tju.edu.cn. Dr. Jun Qing Lu is an associate
professor of physics at East Carolina Univer-
sity; e-mail: luj@ecu.edu.
References
1. P.J. Crosland-Taylor (January 1953). A de-
vice for counting small particles suspended
in a fluid through a tube. Nature, pp. 37-38.
2. W. Dittrich, W. Gohde (March 1969). Im-
pulsfluorometrie bei Einzelezellen in Suspen-
sionen. Z Naturforsch B, pp. 360-361.
3. M.R. Melamed et al (1990). Flow Cytometry
and Sorting. 2nd ed. Wiley-Liss.
4. S.H. Ong et al (October 1987). Development
of an imaging flow cytometer. Anal Quant
Cytol Histol, pp. 375-382.
5. D. Gabor (1948). A new microscopic princi-
ple. Nature, Vol. 161, pp. 777-778.
6. J.Q. Lu et al (April 4, 2005). Simulations of
light scattering from a biconcave red blood
cell using the FDTD method. J Biomed Opt,
024022.
7. R.S. Brock et al (November 2006). Effect
of detailed cell structure on light scattering
distribution: FDTD study of a B-cell with
3D structure constructed from confocal
images. J Quant Spectrosc Radiat Transfer,
pp. 25-36.
8. H. Ding et al (2007). Angle-resolved Mueller
matrix study of light scattering by B-cells at
three wavelengths of 442, 633, and 850 nm.
J Biomed Opt, Vol. 12, 034032.
9. K.M. Jacobs et al (2009). Development of a
diffraction imaging flow cytometer. Opt Lett,
Vol. 34, pp. 2985-2987.
10. K.M. Jacobs et al (2009). Diffraction imag-
ing of spheres and melanoma cells with a
microscope objective. J Biophotonics,
Vol. 2, pp. 521-527.
11. K. Dong et al (2011). Study of cell clas -
s ification with a diffraction imaging flow
cytometer method. SPIE Proc, 7902.
12. J. Neukammer et al (2003). Angular distri-
bution of light scattered by single biological
cells and oriented particle agglomerates.
Appl Opt, Vol. 42, pp. 6388-6397.
13. K. Singh et al (February 2004). Analysis of
cellular structure by light scattering measure-
ments in a new cytometer design based on
a liquid-core waveguide. IEE Proc Nano -
biotechnol, Vol. 151, pp. 10-16.
Flow Cytometry
F
orensic crime shows such as CSI have
taken over TV networks worldwide,
providing viewers with a stylized
glimpse into forensic science. A vital part
of an investigators armory is the forensic
microscope, crucial for inspecting trace
evidence and for making bullet and tool
mark identifications. For a century now in
forensic biology, light microscopy has
been used to identify spermatozoa in
sexual assault cases.
Sadly, real-life crime laboratories often
have hundreds of thousands of cases
in vaults that have not yet been opened
some laboratories can even be years be-
hind, said Richard E. Dick Bisbing, a
forensic scientist for more than 40 years
and formerly with the Michigan State Po-
lice. He is now executive vice president of
McCrone Associates Inc. and an instructor
at Hooke College of Applied Sciences
LLC, both in Westmont, Ill.
There are horror stories about crimi-
nals roaming free because there are inade-
quate resources and too many samples
unworked, he said.
Digital imaging
Consequently, forensic microscopy
makers are looking to advance the devel-
opment of more highly automated systems
so that laboratory technicians can process
samples more quickly, easily, accurately
and less expensively.
Recent advances include combining
optical work with spectroscopy and
spectrophotometry for identifying fibers,
paints and polymers, but the biggest
change has occurred in the past few years:
digital imaging.
Digital imaging via instruments
such as the new Olympus polarized
light microscopes BXiS and BX53
means that microscopic images can be
captured with the touch of a button,
then automatically labeled and archived
together with related sample and analytical
information.
Back when we had to capture images
on film, documentation was often lacking.
Today, most laboratories take photos of
everything because they have high-quality
imaging systems built onto their micro-
scopes, Bisbing said. Thats been a big
change in forensic science in just the past
four or five years.
Thanks to the microscopes coded nose-
piece, all the calculations and magnifica-
tions are input automatically.
What this means in the laboratory
CSI Experts Find Clues
Faster with Microscopy
Tools such as the Olympus BXiS microscope,
when used with Stream software, provide
a comprehensive instrument for analysis of
forensic samples. A cross section of a composite
material is examined and measured using the
Olympus BX51M microscope and Stream
software. Courtesy of Olympus America Inc.
As criminal cases pile up, automation and digital imaging
techniques will help crime scene investigators
close them more quickly.
BY MARIE FREEBODY, CONTRIBUTING EDITOR
ND11_Forensic Feat_Layout 1 10/27/11 10:25 AM Page 35
where imaging is done is a much lower
cost per image, along with the ability to
store multiple images in multiple places
so data is never lost, said Matt Smith,
director of sales and marketing, Scientific
Equipment Group Industrial, of
Olympus America Inc. in Center Valley,
Pa. Hardware is only half a product
now users are really buying an inte-
grated system.
Combining optical techniques with in-
frared spectroscopy allows scientists to
identify paints, adhesive tapes, minerals
and related samples. In addition, Raman
spectroscopy techniques are growing in
popularity.
In some cases, forensic scientists use a
light microscope to make initial compar-
isons and select an appropriate portion of
the sample for analysis, and then use an-
other technique for some of the more
detailed work.
The integration of spectroscopy with
microscopes is expected to become more
refined, as is the streamlined integration
of entire systems, including software,
hardware and storage.
The more efficient and less costly
computer power becomes, the faster
this change will occur, Smith said. In
addition, creating databases that can be
queried easily is an area of importance.
Because photography increasingly is a
part of the work flow, digital camera
technology will continue to develop, with
cameras like the Olympus DP26, DP72
and other cameras that are more accurate,
provide better color rendition and are
push-button easy.
Chromosome sequencing advances
On the biological side, there have been
advances in chromosome sequencing,
and there is progress in performing intra-
cellular work with light microscopy.
Fluorescence and other light microscopy
techniques can now be applied to smaller
samples, down to individual cells.
From the toolmakers and manufactur-
ers perspectives, one of the greatest
changes is that the spectrographic compo-
nent, the optical components and all the
resulting data now can live in the same
Forensic Microscopy
A microscope magnifies cat hairs. Courtesy of McCrone Microscopes & Accessories.
Asbestos fibers are typical specimens for forensic microscopes. Courtesy of Olympus America Inc.
Unitrons comparison forensic microscope was
designed with the help of police organizations,
forensic scientists and educators in the field of
forensic science to inspect ballistics and firearms
and to make tool mark examinations.
Courtesy of Unitron Ltd.
system using the same database, along
with images.
This is powerful because all the
techniques can be integrated on a
single platform such as the BXiS micro-
scope, used with Stream software, to
speed up the work flow for forensic
scientists, Smith said. Olympus also
partners on an OEM basis with some
excellent FTIR [Fourier transform in-
frared] and Raman systems, and all the
data from all of these techniques lives
in a single database.
Polarized light microscopes and other
modular instruments on the market accept
polarized light accessories but also can
be used for phase contrast, fluorescence
and other techniques used by biomedical
researchers and forensic biologists, ac-
cording to Jeffrey McGinn, president
and director of instrument sales at
McCrone Microscopes & Accessories
Inc. in Westmont, a sister company of
McCrone Associates.
Comparison microscopes and stereo-
microscopes also are used in forensic
applications, McGinn said. Additionally,
stereomicroscopes are always sold with
the polarized light systems. The stereos
are used for sample examination with
a larger field of view and the true 3-D
perspective offered by stereoscopic
viewing.
Unitron Ltd. of Commack, N.Y., re-
cently launched a comparison forensic
microscope (CFM) and is working on
approvals to place it into governmental
facilities. The CFM was designed with
the help of police organizations, forensic
scientists and educators in the field of
forensic science to inspect ballistics and
firearms and to make tool mark examina-
tions.
Evidence can be observed simultane-
ously via the comparison bridge that sup-
ports two sets of matched objective steps
on a five-step magnification changer.
The microscope series, equipped with
22-mm field-of-view eyepieces, produces
Forensic Microscopy
erect, unreversed images that move in the
same direction as the specimen for ease
of use. The images can be viewed at
100 percent right or left side, split screen
or superimposed.
I took our CFM to a forensic confer-
ence a few months ago and received noth-
ing but kudos from the state police, sher-
iffs department and local Bureau of
Investigation, said Heston Singh, a busi-
ness developer at Unitron. Our products
are considered mechanically and optically
competitive and are considered far
superior to most of the products coming
out of the Pacific Rim.
Beyond CSI
Its not just crime scene units that make
use of forensics: The FBI and other fed-
eral entities, private firms, private labora-
tories and even industrial laboratories use
forensic instruments. Some industrial
forensic work lies in research and devel-
opment, and some is used to solve prob-
lems relating to litigation and safety be-
cause some events can be understood only
by studying a material closely and identi-
fying its origin or composition.
Hooke College and McCrone Micro-
scopes & Accessories are carrying out
extensive US National Guard training,
working with first responders to teach
them essential techniques to identify
samples and assure safety when working
with unknown and potentially hazardous
materials.
When there is an incident, they show
up on the scene and aid in homeland
defense, McGinn said. We continue
to train these teams, who come to our fa-
cility for basic and advanced microscopy
courses.
Other future developments could come
from projects such as the cooperative
agreement between the National Institute
of Justice and Hooke College, in which
300 trace evidence examiners have been
trained so far in the use of light and elec-
tron microscopes, along with Raman and
infrared spectroscopy.
This is a development that can truly
help advance the practice of forensic
science, Bisbing said.
marie.freebody@photonics.com
There are horror stories about
criminals roaming free because
there are inadequate resources
and too many samples
unworked.
Dick Bisbing, forensic scientist
R
esearchers working with lab-on-a-
chip and cell experiments to date
have relied on general-use micro-
scopes to view the flow of materials
through microchannels and to image mi-
croparticles. These instruments typically
are large and expensive and lack the fea-
tures for viewing live cells and other parti-
cles. Their look-down design inhibits
access for high-voltage electrodes, syringe
pumps and other hardware necessary for
micro- and nanofluidic research and
development experimentation.
A modular, inverted fluorescence syn-
chronized video microscope (Figure 1)
has been developed for imaging micro-
and nanofluidic bioanalyzer prototypes.
The system combines bottom-up viewing
and illumination with a motionless sample
stage, enabling visualization of microsys-
tems without perturbing the fluid flow.
The motionless stage also acts as an opti-
cally accessible lab bench, with unhin-
dered access for external electrical and
fluid connections.
Microanalytical methods hold great
promise for rapid microbial detection,
separation and concentration. Researchers
working with microanalytical methods
require an imaging platform that allows
them to verify in real time the pres-
ence of particles and cells passing through
microchannels or capillaries. Traditional
microscopes, in which the optics are
housed in a stationary head above a mov-
ing stage, have several drawbacks for
these experiments:
The moving stage can easily perturb
the microfluidic system.
The microscopes objective impedes
access to the microfluidic chips,
fluid circuits or cell plates and
other hardware.
Standard illumination can quench
the fluorescent dyes used to view
the particles or microorganisms.
The inverted fluorescence synchronized
video microscope was designed to provide
high-quality images and video of micro-
High-quality video and
images from an inverted
fluorescence synchronized
video microscope show
promise for imaging
micro- and nanofluidic
bioanalyzer prototypes.
Figure 1. The inverted fluorescence synchronized video microscope.
Inset image shows epifluorescence module imaging a micro fluidic
channel from below. Images courtesy of LabSmith Inc.
Inverted Fluorescence Microscopy Aids
Microseparation Studies
BY DR. YOLANDA FINTSCHENKO, LABSMITH INC.,
AND DR. BLANCA H. LAPIZCO-ENCINAS,
TENNESSEE TECHNOLOGICAL UNIVERSITY
nd11_FeatLabsmith_Layout 1 10/27/11 10:26 AM Page 38
and nanosystems. The instruments small
size allows it to be used on a desk or
benchtop. The stage is immobile, leaving
the experimental setup unperturbed by the
imaging process. Images are displayed on
a computer, so there is no need for over-
head viewing optics.
Figure 2 shows an exploded view of the
microscopes major components. The op-
tics are contained within the camera mod-
ule. An LED ring illuminator module can
provide external illumination at a variety
of wavelengths for fluorescence and dark-
field microscopy. Stroboscopic illumina-
tion reduces quenching of fluorescent dyes.
The camera and illumination modules both
attach to an X-Y stage that moves beneath
a stationary stage top. This design lets re-
searchers construct microsystems directly
on the microscope and allows unimpeded
connection to other equipment.
An epifluorescence camera module can
provide greater wavelength selectivity for
improved signal-to-noise. This module
contains illumination and filters as well
as the camera. Light returned from the
sample follows a common optical path.
The microscopes operating software
also can record pictures or movies and
can insert velocity probes for particle
image velocimetry experiments.
Bioanalytical efficiency
An example of the efficacy of the mi-
croscope is in the research of Dr. Blanca
H. Lapizco-Encinas, associate professor
of chemical engineering and director of
the Microscale Bioseparations Laboratory
at Tennessee Technological University in
Cookeville. Her team is studying an im-
proved method for single-step separation
and concentration of macromolecules,
microorganisms and nonorganic particles.
The method, insulator-based dielec-
trophoresis (iDEP),
1,2
is a variant of dielec-
trophoresis, a process in which nonuniform
electric fields concentrate and separate par-
ticles.
3
In the iDEP technique, two remote
electrodes apply a direct current across a
microchannel filled with a conductive
electrolyte and an array of insulating
structures. The structures do not conduct
current and thus create the nonuniform
electric fields necessary for DEP.
By varying experimental parameters
and the shape, size, spacing and orienta-
tion of the insulator posts, particles and
cells of specific sizes can be trapped, sep-
arated and concentrated in seconds or min-
utes, rather than the hours or days required
by traditional concentration techniques.
Because iDEP microdevices are inexpen-
sive and relatively simple to fabricate, the
method holds significant promise for
miniaturized high-throughput sampling
systems. The electric field gradient is
easily controlled across the entire volume,
enabling scalability.
For the iDEP method to be widely im-
plemented, it is necessary to understand
Figure 2. Exploded view of the microscopes major components.
Figure 3. Schematic representation of the microfluidic channel setup for studying the parameters of the iDEP
method, (a) side view and (b) top view.
Figure 4. Experimental setup shows
vacuum manifold and a circular
glass microfluidic chip on the stage
of the microscope.
the conditions that result in the best trap-
ping of various-size particles and organ-
isms. To that end, Lapizco-Encinas and
her team developed extensive, predictive
models encompassing a range of experi-
mental conditions. To corroborate the
models, a set of iDEP experiments was
conducted, in which a microchannel was
fabricated containing an array of cylindri-
cal, insulating posts, 200 m in diameter
and spaced on 250-m centers (Figure 3).
A DC field is applied both to trap the par-
ticles and to drive the fluid flow through
the channel by means of electro-osmosis.
4
Real-time examination of events in the
microchannel was required over the course
of the experiments to verify the predictive
models. An inverted fluorescence video
microscope was selected as the imaging
platform for the iDEP experiments.
The instrument was perfectly suited to
our work, said Hctor Moncada Hernn-
dez, a senior PhD student in Lapizco-
Encinas laboratory. We were able to
construct our experimental setups directly
on the microscope, as if it were a table.
Since we were working with voltages of
1000 V or more, it was also important that
we did not have a microscope objective or
other metal parts in the field that could
ground our electrodes.
Figure 4 shows the setup for the iDEP
experiments, in which a vacuum manifold
and a circular glass microfluidic chip are
placed atop the stationary stage of the
microscope.
In a first set of studies, a sample of
fluorescent 1-m microspheres was intro-
duced at the inlet of the microchannel and
a voltage applied to drive the fluid flow
through the channel (Figure 5). Images
and video of the fluid flow allowed the
researchers to fine-tune the voltage, as
well as the pH and conductivity of the
suspending buffer, to maximize particle
trapping (Figure 5a).
In a second set of experiments, a sam-
ple containing E. coli was introduced into
the microchannel. The cells were labeled
with SYTO 9 fluorescent dye for visibility.
Whereas the illuminator of a standard
microscope can quench the dye, leaving
the cells indiscernible, the stroboscopic
illuminator of the inverted fluorescence
video microscope minimizes quenching,
enabling high particle visibility over
longer experiments.
In addition, the microscopes epifluores-
cence camera module was used to improve
visibility. The epifluorescence capability
Fluorescence Microscopy
Figure 5. A DC potential of 800 V is applied
across a microchannel with suspended 1-mm
microspheres. The particles appear in bright
white between the insulator posts. Researchers
used images such as these to fine-tune parame-
ters to maximize particle trapping. (a) Particles
are being successfully immobilized by employ-
ing a suspending medium with conductivity of
100 s/cm and pH of 6. (b) Immobilization
decreases when pH is increased to 9. (c) Immo-
bilization decreases when conductivity is de-
creased to 25 s/cm.
4
Figure 6. E. coli cells trapped at a potential of 1000 V, viewed with a 4 objective. (a) Image taken using
standard LED illumination. (b) Same image, taken with the epifluorescence module, which improves imaging
of low concentrations.
Figure 7. Dielectropherogram shows peaks of concentrated E. coli and S. cerevisiae cells as a function of
time and applied DC voltage. Simultaneous concentration and separation in <2 min was visualized with the
microscope by fluorescence measurements.
5
helped us to see low concentrations of
particles that we may have missed using
standard illumination, Lapizco-Encinas
said. Figure 6 shows E. coli cells trapped
at a potential of 1000 V, as imaged with
standard illumination (left) and with
improved visibility resulting from the
use of epifluorescence (right).
A third set of studies shows the great
potential of the iDEP method for rapid
microorganism concentration and separa-
tion.
5
Water from a pond was filtered to
remove larger particles, then spiked with
labeled E. coli and Saccharomyces cere-
visiae yeast cells. An electric field was
applied to immobilize and concentrate
both types of cells. The field was then
lowered to release the bacteria, and 30
seconds later, the field was lowered
again to release the yeast cells.
Fluorescence measurements made with
the microscope allowed researchers to plot
the passage of high concentrations of each
type of cell as clearly separated events.
Figure 7 shows a dielectropherogram of
the events, in which an electric field gradi-
ent eluted the concentrated cells from the
microchannel.
A further advantage of the inverted
fluorescence microscope is its integrated
software for capturing images and video
of fluid flow and analyzing the results.
The software includes facilities for auto-
matically triggering video capture based
on the presence of microparticles or other
events. This capability ensures that critical
events are not missed, even over the
course of hours- or days-long experiment
runs. Particle imaging velocimetry tools
are integrated with the microscope soft-
ware, which allows researchers to easily
map the velocity of the flow through
microfluidic channels.
4
The inverted fluorescence synchronized
video microscope addresses the require-
ments of researchers working with micro-
or nanofluidic experiments. It brings to-
gether the features required for microflu-
idics work, including a stationary stage,
high-quality imaging, and software to cap-
ture publication-quality images and video.
Meet the authors
Dr. Yolanda Fintschenko is the director of new
technologies, marketing and sales at LabSmith
Inc. in Livermore Calif.; e-mail: yfintschenko
@labsmith.com. Dr. Blanca H. Lapizco-Encinas
is associate professor of chemical engineering
and director of the Microscale Bioseparations
Laboratory at Tennessee Technological
University.
References
1. E.B. Cummings et al (2003). Characteriza-
tion of electrokinetic mobility of microparti-
cles. Anal Chem, Vol. 75, pp. 4724-4731.
2. B.H. Lapizco-Encinas et al (2004). Dielec-
trophoretic concentration and separation of
live and dead bacteria in an array of insula-
tors. Anal Chem, Vol. 76, pp. 1571-1579.
3. H.A. Pohl (1951). The motion and precipita-
tion of suspensoids in divergent electric
fields. J Appl Phys, Vol. 22, pp. 869-871.
4. J.I. Martnez-Lpez et al (2009). Characteri-
zation of electrokinetic mobility of micropar-
ticles in order to improve dielectrophoretic
concentration. Anal Bioanal Chem, Vol. 394,
pp. 293-302.
5. H. Moncada Hernndez et al (2010). Simul-
taneous concentration and separation of
microorganisms: insulator-based dielectro -
phoretic approach. Anal Bioanal Chem,
Vol. 396, Issue 5, pp. 1805-1816.
Fluorescence Microscopy
2012 OSA
OPTICS &
PHOTONICS
CONGRESS Call for Papers
Visit: www.osa.org/biomedicaloptics for more information
Biomedical Optics and
3-D Imaging
S S E R ONG C
S C I ON T HO P
S & C I T P O
A S O 12 0 2

d i B

i t l O i d

d

o YYo itt m b u S
a il 2 M r p 9 A 2
n D o t il i H m a i M
ag m I D - 3
d e m o i B

w o r N e ap r P u o
2 1 y 20 a
, F i m a i n M w o t n w o n D
ng i ag
c i t p l O a c i d

! ww!
A S , U a d i r o l
d n s a

n D
or osa. . w w w isit: V
io t a r t s i g Re - e r P
a e n D io s s i m b u S
g O n i r e e n i g n l E a c i d e m o i B
l , A o r p s a i : D t i d e r o c t o h P

0
f g/biomedicaloptics or
ch 2 r a 9 M : 1 e n i l d a e n D
0 r 2 e b m e c e 4 D : 1 e n i l d
. 6 3 : , 5 6 0 0 , 2 e n i l n g O
n m o i t a t i c x n e o t o h P - i t l u . M l t a ; e o t r e b

12
tion orma e inf or mor f
0
1 1 0
. y p o c s o r c i n m

Revolution DSD
Andors Revolution DSD is an innovative spinning disk technology that
brings an affordable confocal solution to your laboratory, offering you
less dependency on laser-based solutions often restricted to core facilities.
While laser-free, the Revolution DSD can still achieve the optical sectioning
you expect of a complex laser scanning confocal system, but with low
maintenance costs.
+44 28 9023 7126
marketing@andor.com
www.andor.com/dsd
See more new products at Photonics.com
Its easy to find the latest products on our website Photonics.com.
Just click on the menu marked PRODUCTS on the navigation bar
(under the logo) to find new products almost every day.
When people ask, Whats new? tell them to go to:
Photonics.com/Products.
(413) 499-0514
photonics.com
advertising@photonics.com
ND11_Spotlight_Layout 1 10/27/11 11:30 AM Page 42
BREAKTHROUGHPRODUCTS
Fiber-Coupled Multibar Modules
Dilas is offering higher-power fiber-coupled multibar
laser modules operating at 1940 nm. The high-
brightness, high-power modules feature a compact
footprint and deliver 30-W output through a 600-m
core diameter fiber with a numerical aperture of 0.22
and wall-plug efficiency of >10%. The designs are
based on industry-standard conductively cooled
bars that are optically stacked and polarization-coupled. The modules require
only industrial water for cooling. Custom wavelengths are available upon request.
The modules are suitable for direct diode applications such as medicine, defense
and welding of transparent plastics.
Dilas
sales@dilas.com
532-nm Laser
Laser Quantum has enhanced its Excel, a
continuous-wave 532-nm laser that outputs
up to 2 W while maintaining noise at <0.5% rms.
Compact and robust in design, its new intelligent
smd12 power supply allows operation in power
or current mode, with power stability of <0.4%.
The lasers low requirements for temperature
control and long warranty periods make it
suitable for scientific applications and integration
into other instruments. Applications include
particle image velocimetry, ophthalmology and microscopy.
Laser Quantum
sales@laserquantum.com
Polarization Imaging
Bossa Nova Technologies LLC has released a full Stokes polariza-
tion imaging camera. Salsa performs live measurement of the full
Stokes vector, including the fourth parameter S3, related to circular
polarization. Dedicated software provides live images of any derived
parameters, such as the degrees of linear and circular polarization,
the angle of polarization and the ellipticity angle. In-house calibration
enables accurate polarization measurements below 1% rms deviation.
Measuring 3.2 3.2 4 in., the robust camera features a 1-mega -
pixel, 12-bit CCD sensor, standard C-mount lenses and a proprietary
fast polarization modulator. The camera operates at up to 12 fps
in polarization imaging mode and is suitable for use in a variety
of applications, including biomedical imaging.
Bossa Nova Technologies LLC
info@bossanovatech.com
Digital Microscope Camera
For live-cell imaging applications, Leica Microsystems has unveiled the DFC365 FX digital microscope
camera. To document live cells and rapidly fading fluorescence specimens, it combines high-quality
imaging with high temporal resolution for rapid time-lapse recordings. It enables researchers to
work efficiently, even with weakly fluorescing specimens. Equipped with a sensitive CCD sensor with
6.45-m pixels and active Peltier cooling, it is suitable for applications ranging from basic fluorescence
documentation to total internal reflection fluorescence, Frster resonance energy transfer and struc-
tured illumination. It operates at acquisition rates of 21 fps at full resolution. In overlapping mode,
an image can be captured while the previous image is still being read out. Data is transferred to the
PC via a FireWire-B interface.
Leica Microsystems
valerie.nicolas@leica-microsystems.com
Photomultiplier Tubes
Hamamatsu Corp. has re-
leased the H11451 series
linear array photomultiplier
tube (PMT) modules. The
detectors offer high cathode
sensitivity, high gain and
high-speed response, with
fast rise time and a narrow
transit time spread. They
contain a multichannel PMT,
along with a preamplifier and
a high-voltage power supply
circuit. Applications range
from biomedical fluorescence
and laser scanning detection
to analysis, including spec-
tros copy and environmental monitoring. The series features eight channels
in a linear format, with each channels photosensitive area measuring
2 2.5 mm. The modules are characterized by good anode uniformity,
low crosstalk and a rise time of 0.7 ns. Two spectral response ranges
are available: 300 to 880 nm (peak at 420 nm) and 300 to 920 nm (peak
at 630 nm).
Hamamatsu Corp.
usa@hamamatsu.com
Fluorescence Illumination
For fluorescence microscopy and live-cell imaging
applications, Lumen Dynamics Group Inc. has un-
veiled the X-Cite XLED1, a high-power LED illumina-
tion solution offering advanced triggering capabilities
that enable precise time control for high-speed image
automation. It provides instant switching of wave-
lengths and fast LED pulsing for greater data integ -
rity. The system is used for excitation or photolysis.
The touch-screen control panel provides a simple
and precise method to control the device and to save
protocols for multiple experiments. With optimized
excitation through high-power LEDs via narrow
spectral profiles, the XLED1 delivers maximum
illumination right at the sample plane. Providing
fine light intensity control, the system gives research -
ers full control for achieving optimal imaging.
Lumen Dynamics Group Inc.
ada.nowensky@ldgi.com
ND11_Breakthrough Leads_Layout 1 10/27/11 11:48 AM Page 43

Laser Platform
Vortran Laser Technology Inc.s Stradus laser
is compatible with Manager open source
confocal software that controls confocal mi-
croscope systems. The software allows the
user to perform common microscope image
acquisition protocols such as time lapses,
multichannel imaging, Z-stacks and other
combinations of techniques. It works with
microscopes from Leica, Nikon, Olympus
and Zeiss. The flexibility of the laser module
allows it to function well with this software.
The Vortran Stradus-GUI software gives com-
plete control of the laser to the user, who can
adjust power, monitor key parameters and
perform custom program scripts.
Vortran Laser Technology Inc.
sales@vortranlaser.com
36-mm Linear Actuator
Haydon Kerk Motion Solutions, a division of
Ametek Inc., has unveiled a new version of
its 36-mm G4 linear actuator. Included is an
adapter plate that allows the smaller 36-mm-
outer-diameter unit to replace 42- and 46-mm
units in existing applications. The company
says that the performance of the device ap-
proaches that of the larger 42- and 46-mm
models from various manufacturers. The
integrated adapter plate uses the same bolt
pattern and pilot surface, allowing customers
to replace existing 42- and 46-mm units easily.
Advantages of substituting include a smaller
linear actuator body, which is important when
space is at a premium, and a lower cost sys-
tem. The 36-mm model is suitable for medical
equipment and laboratory instrumentation
applications.
Haydon Kerk Motion Solutions
info@haydonkerk.com
Machine Vision Cameras
Imperx Inc. supports a wide range of high-res-
olution video-capture applications with two
29-megapixel B6620 cameras. High resolution
is important for video-capture applications
where large areas must be viewed in fine
detail. Shutter speeds of up to 1.25 10
5
s
freeze rapid motion. Frame rates up to 2.4 fps
follow rapidly evolving scenes. The standard
F-mount enables fitting the cameras to almost
any optical system requirement. The cameras
ability to image at light levels as low as 1 lx
(f/1.4) allows capturing dimly lit scenes. The
cameras feature compact size, light weight, a
wide operating temperature range, low power
consumption and a high shock rating. Applica-
tions include biomedical research.
Imperx Inc.
sales@imperx.com
Diode Laser Stacks
Dilas has released high-duty-cycle vertical
diode laser stacks that enable long pulse
lengths for hair removal and high-energy
applications. The compact gold-tin-soldered
vertical stacks are designed for operation in
medical applications with low cooling capacity
and for high-energy laser applications requir-
ing high pulse energy in a small, dense pack-
age. From the parameter field defined by duty
cycle, power and pulse length, the diode
lasers start at 200 W/808 nm and 300 W/940
nm at 2% duty cycle with 2-ms pulse duration,
up to 60 W per bar at 808 nm and 175 W per
bar at 940 nm at 20% duty cycle with 75-ms
pulse duration. Custom options are available.
Dilas
sales@dilas.com
CCD Cameras
Princeton Instruments PyLoN series con-
trollerless, cryogenically cooled CCD cameras
are designed for quantitative spectroscopy
applications that demand high sensitivity. The
PyLoN:100 and PyLoN:400 dual-amplifier cam-
eras use 1340 100- and 1340 400-pixel
sensors, respectively. In creating the new plat-
form, the company redesigned its Spec-10
family of cameras to remove the external con-
troller, increasing experimental flexibility and
improving the ultralow-noise electronics.
Liquid nitrogen cooling virtually eliminates
dark current, and indium metal seals enhance
vacuum longevity. Binning noise has been
reduced. Both cameras provide software-
selectable gain that permits operation in
high-sensitivity mode for Raman or single-
molecule spectroscopy, and in high-capacity
mode for fluorescence spectroscopy. The
PyLoN:100 is suitable for rapid spectral acqui-
sition, and the PyLoN:400 for multiplexed
spectroscopy.
Princeton Instruments
info@princetoninstruments.com
Microstructures
Available from IMT Mask und Teilungen AG
are microstructures in sizes down to 0.150 m
in combination with optical coatings on glass
and other substrates. They are customized for
original equipment manufacturer solutions
such as reticles, lab-on-chip components, light
valves, beamsplitters, microapertures, masks,
encoder discs and scales, calibration plates,
targets and mirrors. The microstructures and
optical coatings are manufactured according
to customer specifications in quantities from
prototyping to large-scale manufacture of
several hundred thousand per year. Suitable
for use in applications in bioscience, screen-
ing, optical devices, biophotonics, detectors,
medical technology, microfluidics, machine
vision sensors and metrology, the products
are available in dimensions from 0.8 mm in
diameter to 24 32-in. plates.
IMT Mask und Teilungen AG
info@imtag.ch
Spectroradiometer
International Light Technologies has released
the ILT950 portable spectroradiometer. The
high-speed, high-precision turnkey instrument
is designed for standard irradiance and color
measurements in a production or laboratory
p BREAKTHROUGHPRODUCTS
nd11_NewProds_Layout 1 10/27/11 12:00 PM Page 44
environment. Typical applications include
colorimetry, characterization of UV curing sys-
tems, photostability testing, solar simulation,
photobiology, photochemistry and LED illumi-
nation. Proprietary SpectriLight III software
provides easy setting of all operating controls.
It has an integrated data analysis package
that renders spectral analysis fast and simple.
Wavelength range, integration time, scan aver-
age and other controls can be set through
pop-up windows, menus and tool bars. Ab-
solute irradiance and chromaticity are calcu-
lated instantly. Color data is verified against
NIST-traceable standards. The software also
offers enhanced scaling and zooming features,
along with movable vertical cursors.
International Light Technologies
ilsales@intl-lighttech.com
Ceramic Motor Stage
Nanomotion has added a Z-wedge vertical
stage to its FB range of compact nanometer-
level micropositioning mechanics. The stages
complement a modular and compact range of
linear, rotary and goniometric stages that to-
gether provide multiaxis and nanometer-level
translation in all six degrees of freedom. The
FBZ075-10 has a classic upper and lower
wedge design featuring a precision-grade
cross roller bearing arrangement that main-
tains stage stiffness, with a side-mounted
ceramic motor driving the bottom wedge via
a ceramic strip. The FB range uses the com-
panys HR series high-force ceramic motors,
providing a wide dynamic speed range from
a few microns per second up to 250 mm/s,
with high acceleration. The technology has
applications in biomedical systems.
Nanomotion
nano@nanomotion.com
GHz Photoreceiver
New Focus, a Newport Corp. brand, intro-
duces the GHz Nirvana, a high-speed photo -
receiver with automatic signal balancing. It
offers high bandwidth and fiber coupling. The
autobalance feature eliminates background
noise from dynamically changing systems, in-
cluding thermal drift and wavelength depend-
ence, yielding perfect power balance between
the reference and signal beams. It eliminates
the need to optically balance the detectors
and enables measurement systems that can
accommodate drift over time. The user can
balance in optical and manual modes. The
receiver improves the performance of swept-
source OCT using high-speed balanced and
autobalanced detection. In this application,
the balanced detection allows subtraction of
all common-mode noise and improves the
ability to resolve low-scattering features
deep in the sample.
New Focus
malcolm.minty@newport.com
Compact Focus Module
New Scale Technologies Inc. has upgraded
the lifetime specification of its miniature M3-F
focus module and provided new repeatability
data for it. The module replaces fixed lens
holders in compact board cameras for biomet-
ric detection and medical diagnostics applica-
tions. The new lifetime is >2 million cycles
mean time between failure in a fixed orienta-
tion or >500,000 cycles in random orientation.
Position repeatability is 5 m unidirectional
and 20 m bidirectional, demonstrating high
performance for high-resolution imaging
applications. Image quality is directly propor-
tional to lens position repeatability with auto-
focus algorithms. With a footprint of 20 22
16 mm and no external control board, the
M3-F easily replaces fixed lens holders over
CCD or CMOS image sensors without adding
to the camera size.
New Scale Technologies Inc.
sales@newscaletech.com
EMCCD Microscopy Camera
For high-sensitivity low-light imaging, QImag-
ing has released the Rolera Thunder electron-
multiplying CCD (EMCCD) microscopy camera.
Its quantitative capability with linear EM gain
BREAKTHROUGHPRODUCTS p
and high frame rates renders advanced imag-
ing accessible to more researchers. Designed
for quantitative low-light fluorescence micros-
copy, the camera is suitable for life sciences
applications including superresolution sto-
chastic optical reconstruction microscopy im-
aging, photoactivated localization microscopy
imaging, single-molecule fluorescence and
fluorescence recovery after photobleaching.
The camera features a back-illuminated 16-
m-pixel, 512 512 EMCCD sensor for >90%
peak quantum efficiency, read noise of <1 e
and operation at >385 fps with binning. Be-

cause of its high sensitivity, very short expo-
sure times are possible, enabling rapid data
acquisition and greater temporal resolution.
QImaging
erasmussen@qimaging.com
Infrared AR Coatings
Proteus, Artemis Optical Ltd.s high-durability
infrared antireflection (AR) coatings, deliver
high transmission over a specified waveband
that increases the systems signal-to-noise
ratio and improves the identification range.
They are designed to meet MIL-C-48497 Para
4.5.5.1, extending system life and operational
performance. They are nonradioactive, which
reduces export restrictions, and because they
are already developed for a wide range of
infrared substrate materials, lead time and
development costs are reduced. Customers
include manufacturers of thermal imaging
cameras and systems. Thermal imaging
allows images of heat signatures to be dis-
played on a monitor or other devices. This
technology has growing application in a
variety of areas, including tumor detection
and photonics research.
Artemis Optical Ltd.
proteus@artemis-optical.co.uk
High-Speed Cameras
Vision Research, a business unit of the Materi-
als Analysis Div. of Ametek Inc., has intro-
duced the Phantom v1210 and v1610 1-
megapixel digital high-speed cameras, which
feature high-definition and wide-screen 1280
800-pixel CMOS sensors. The Phantom
v1610 acquires more than 16,000 fps at full
resolution and up to 1 million fps at reduced
resolution. Capturing high-resolution images
at ultrahigh speeds, the cameras benefit re-
searchers who need to see phenomena invisi-
ble to the human eye. The cameras are based
on sensors that offer 28-m pixels that pro-
duce high sensitivity when shooting in low
light. The v1210 can be configured with 12, 24
or 48 GB of memory, and the v1610, with 24
or 96 GB. The memory can be segmented into
as many as 63 partitions.
Vision Research
phantom@visionresearch.com
Laser Alignment Tuning Kits
Newport Corp. has released its LAKIT series
laser power alignment tuning kits. Suitable
for use in research setups, the kits include
the new 1917-R laser power meter and a wide
selection of optical detectors, making it easier
for researchers to match the laser type. Each
kit contains an optical power meter, a detector
and the corresponding mounting assembly.
The kits use silicon, germanium or thermopile
detectors. They operate in wavelength ranges
from 200 nm to 10.6 m, with up to 100-W
average power. The 918D series, designed for
low-power lasers (<2 W/cm
2
), operates in the
visible and infrared spectra. The 818P series
thermopile detectors are suitable for measur-
ing the average power for high-power lasers
that operate in both continuous-wave and
pulsed modes.
Newport Corp.
jay.jeong@newport.com
Automated Work Cell
Nordson Asymtek has introduced an auto-
mated work cell for film-frame wafer-level
packaging applications. Intended for cus-
tomers who want to buy their dispensing
and handling equipment from a single vendor,
the MH-910W integrates wafer dispensing and
handling in one automated work cell to mini-
mize footprint and maximize throughput.
The work cell same-side loader/unloader
integrated with the Spectrum S-920N dispenser is designed for 150-mm
film-frame wafer processing and is flexible enough to support various
sizes. Advanced process controls are built in to ensure smooth, error-
free transport and dispensing. The MH-910W film-frame loader/un-
loader and the S-920N dispensing system are fully enclosed, with
interlocked doors and windows for operator safety and part security.
Applications include biochip reagent jet dispensing in life sciences
applications.
Nordson Asymtek
info@nordsonasymtek.com
Laser Diode Drivers
The PPS series analog laser diode drivers/controllers manufactured by
Portable Power Systems Inc. can operate in pulsed or continuous-wave
mode. Depending upon the unit, voltages range from 0 to 250 V, and
currents range from 0 to 300 A. Rise times of 15 s and pulse widths
of 25 s are possible. Most units house a universal input power supply
that accepts voltages of 85 to 264 VAC. Higher-power units can work
with an external power supply to reach higher power levels. The cur-
rent can be controlled by a potentiometer on the front panel or by
accepting a 0 to 10-V analog signal via the rear panel. Medical applica-
tions include photodynamic therapy and vein cauterization.
Portable Power Systems Inc.
sales@pwrsys.com
BREAKTHROUGHPRODUCTS p
Advertise in BioPhotonics
Lasers, optics, imaging, microscopy and spectroscopy
for the life sciences in every issue.
January
Content focus: Dentistry
Sneak Preview: SPIE Photonics West
Show distribution: SPIE BiOS, Photonics West
February
Content focus: Pharmaceuticals
Show distribution: PITTCON,
Biophysical Society Annual Meeting
March
Content focus: Cancer
April
Content focus: Ophthalmology
Sneak preview: CLEO
Show distribution: OSA Congress:
Biomedical Optics & 3-D Imaging
CLEO, SMEs mfg4
Contact your sales representative or:
sales@photonics.com (413) 499-0514
Shine the spotlight on your Breakthrough Product in a display ad
in BioPhotonics. Contact Kristina Laurin at (413) 499-0514 or at
advertising@photonics.com
APPOINTMENTS
DECEMBER
Optics in Cardiology (Dec. 1-2) Rotterdam,
Netherlands. Contact Mieke Pruijsten, Erasmus
MC, +31 10 704 4030; m.pruijsten@erasmus
mc.nl; www.opticsincardiology.org.
Functional Optical Imaging 2011 (Dec. 3-4)
Ningbo, China. Contact Laura Nightingale,
University of Nottingham, +44 115 84 68504;
ibios@nottingham.ac.uk; www.nottingham.
ac.uk/ibios.
2011 ASCB Annual Meeting (Dec. 3-7) Denver.
Contact American Society for Cell Biology,
+1 (301) 347-9300; ascbinfo@ascb.org;
www.ascb.org.
SPIE Smart Nano+Micro Materials
and Devices (Dec. 5-7) Melbourne, Australia.
Contact SPIE, +1 (360) 676-3290; customer
service@spie.org; spie.org.
First EOS Topical Meeting on Micro- and
Nano-Optoelectronic Systems (Dec. 7-9)
Bremen, Germany. A European Optical Society
event. Contact Julia Dalichow, EOS Events
& Services GmbH, +49 511 277 2673; bremen
@myeos.org; myeos.org/events/bremen2011.
Scientific Meeting of Photonics4Life (Dec. 7-9)
Karlsruhe, Germany. Contact Juergen Popp,
+49 3641 206 300; juergen.popp@ipht-jena.de;
www.photonics4life.eu.
Society of Electron Microscopy Technology
Meeting 2011 (Dec. 12) London.
Contact David McCarthy, University of
London, +44 20 7753 5806; david.mccarthy@
pharmacy.ac.uk; www.semt.org.uk.
JANUARY
Live-Cell Imaging (Jan. 12-13) San Francisco.
Collocated with High-Content Analysis.
Contact Julia Boguslavsky, Cambridge
Healthtech Institute, + 1 (781) 972-5400;
juliab@healthtech.com; www.healthtech.com.
SPIE Photonics West (Jan. 21-26) San Fran-
cisco. Includes the symposia BiOS Biomed-
ical Optics; LASE Lasers and Applications;
OPTO Optoelectronic Materials and Devices;
and MOEMS MEMS Micro and Nanofabri-
cation. Contact SPIE, +1 (360) 676-3290;
customerservice@spie.org; spie.org.
IS&T/SPIE Electronic Imaging (Jan. 22-26)
Burlingame, Calif. Contact SPIE, +1 (360) 676-
3290; customerservice@spie.org; spie.org.
Lasers, Sources and Related Photonics
Devices: OSA Optics and Photonics Congress
(Jan. 29-Feb. 3) San Diego. Includes the
meetings Advanced Solid State Photonics
(ASSP); Advances in Optical Materials (AIOM);
Fiber Laser Applications (FILAS); and Laser
Applications to Chemical, Security and
Environmental Analysis (LACSEA). Contact
Optical Society of America, +1 (202) 223-8130;
info@osa.org; www.osa.org.
Third Asia-Pacific Optical Sensors
Conference (APOS 2012) (Jan. 31-Feb. 3)
Sydney, Australia. Contact Conference
Secretariat, secretariat@apos2012.com;
www.apos2012.org.
FEBRUARY
BIOSTEC 2012; Fifth International Joint
Conference on Biomedical Engineering
Systems and Technologies (Feb. 1-4)
Vilamoura, Portugal. Contact BIOSTEC
Secretariat, +351 265 100 033; biostec.
secretariat@insticc.org.
SPIE Medical Imaging (Feb. 4-9) San Diego.
Contact SPIE, +1 (360) 676-3290; customer
service@spie.org; spie.org.
APMC-10, ICONN 2012 and ACMM-22
(Feb. 5-9) Perth, Western Australia.
Comprises the 10th Asia-Pacific Microscopy
Conference, the International Conference
on Nanoscience and Nanotechnology and
the 22nd Australian Conference on Microscopy
and Microanalysis. Contact Event Manager,
EECW Pty Ltd., +618 9389 1488; registration
@eecw.com.au; www.iconn-2012.org or
www.apmc-10.org.
MMVR19/NextMed
(Medicine Meets Virtual Reality) (Feb. 9-11)
Newport Beach, Calif. Contact Aligned
Management Associates Inc., +1 (805)
534-0300; mmvr19@nextmed.com;
www.nextmed.com.
Biophysical Society 56th Annual Meeting
(Feb. 25-29) San Diego. Contact Biophysical
Society, +1 (240) 290-5600; society@bio
physics.org; www.biophysics.org/2012
meeting.
Eighth EOS Topical Meeting on Diffractive
Optics (DO 2012) (Feb. 27-March 1) Delft,
Netherlands. A European Optical Society
event. Contact Renate Rebmann, EOS
Events & Services GmbH, +49 511 277 2676;
do2012@myeos.org; www.myeos.org/events/
do2012.
MARCH
Pittcon 2012 Laboratory Science
Equipment Conference and Exposition
(March 11-15) Orlando, Fla.
Contact The Pittsburgh Conference,
+1 (412) 825-3220 or +1 (800) 825-3221;
info@pittcon.org; www.pittcon.org.
SPIE Smart Structures/NDE (March 11-15)
San Diego. Contact SPIE, +1 (360) 676-3290;
customerservice@spie.org; spie.org.
Research in Optical Sciences Congress:
OSA Optics and Photonics Congress
(March 19-21) Berlin. Includes High Intensity
Lasers and High Field Phenomena (HILAS);
International Conference on Ultrafast
Structural Dynamics (ICUSD); and Quantum
Information and Measurement (QIM).
Contact Optical Society of America, +1 (202)
223-8130; info@osa.org; www.osa.org.
Laser Optics Berlin International Trade
Fair and Congress for Optical and Laser
Technologies (March 19-21) Berlin. Contact
Kerstin Kube-Erkens, Messe Berlin GmbH,
+49 30 3038 2159; laser-optics@messe-
berlin.de; www.laser-optics-berlin.de.
CALL FOR PAPERS
CLEO: 2012 May 6-11
Deadline: Abstracts and summaries, December 5, 17:00 GMT
San Jose, Calif. Organizers of CLEO Laser Science to Photonic
Applications (Conference on Lasers and Electro-Optics) invite abstracts
for the meetings CLEO:QELS Fundamental Science (Quantum
Electronics and Laser Science); CLEO: Science and Innovations,
which will address biophotonics and optofluidics topics, among
others; and CLEO: Applications and Technology, which will consider
biomedical aspects.
Contact: Optical Society of America, CLEO Management
+1 (202) 416-1907 custserv@osa.org
www.cleoconference.org
Biomedical Optics and 3-D Imaging April 29-May 2
Deadline: Abstracts, December 14, noon EST (17:00 GMT)
Miami. Papers are sought for Biomedical Optics and 3-D Imaging:
OSA Optics and Photonics Congress, which will include the topical
meetings Biomedical Optics, and Digital Holography and 3-D Imaging.
Topics to be considered include in vivo imaging and therapies,
holographic interferometry for deformation or contour measurement,
3-D optical remote sensing and 3-D optical image processing.
Contact: Optical Society of America +1 (202) 223-8130
info@osa.org www.osa.org
Focus on Microscopy 2012 April 1-4
Deadline: Abstracts, January 10
Singapore. Submissions are encouraged for FOM 2012 Focus on
Microscopy. Key topics to be discussed include 3- and 4-D live-cell
and tissue imaging; time-resolved fluorescence; coherent, nonlinear
microscopies; multidimensional fluorescence and Raman spectroscopy
imaging; bio- and nanomaterials, biosensors; OCT and endoscopy;
and 3-D image processing and visualization for multidimensional data.
The conference also covers advanced fluorescence labeling techniques
for confocal and multiphoton imaging of live biological specimens.
Contact: FOM 2012 info2012@focusonmicrosopy.org
www.focusonmicroscopy.org
For complete listings, visit
www.photonics.com/calendar
nd11Appointments_Layout 1 10/27/11 10:29 AM Page 48
49
ADVERTISERINDEX
Photonics Media Advertising Contacts
Please visit our client service
website Photonics.com for all
your marketing needs.
Ken Tyburski
Director of Sales
Voice: +1 (413) 499-0514, Ext. 101
Fax: +1 (413) 443-0472
ken.tyburski@photonics.com
New England, Southeastern US & FL,
Rocky Mountains, AZ, NM, Midwest
Rebecca L. Pontier
Associate Director
Voice: +1 (413) 499-0514, Ext. 112
Fax: +1 (413) 443-0472
becky.pontier@photonics.com
NY, NJ & PA, Eastern Canada
Timothy A. Dupree
Regional Manager
Voice: +1 (413) 499-0514, Ext. 111
Fax: +1 (413) 443-0472
tim.dupree@photonics.com
Northern CA, Pacific Northwest,
AK, NV, Yukon & British Columbia
Joanne C. Gagnon
Regional Manager
Voice: +1 (413) 499-0514, Ext. 226
Fax: +1 (413) 443-0472
joanne.gagnon@photonics.com
Southern CA, Central CA & HI
Tracy L. Reynolds
Regional Manager
Voice: +1 (413) 499-0514, Ext. 104
Fax: +1 (413) 443-0472
tracy.reynolds@photonics.com
Europe, Israel & South Central US
Owen Broch
Regional Manager
Voice: +1 (413) 499-0514, Ext. 108
Fax: +1 (413) 443-0472
owen.broch@photonics.com
Austria, Germany & Liechtenstein
Olaf Kortenhoff
Voice: +49 2241 1684777
Fax: +49 2241 1684776
olaf.kortenhoff@photonics.com
Asia (except Japan)
Hans Zhong
Voice: +86 755 2157 3066
Fax: +86 755 2872 6973
hans.zhong@yahoo.com.cn
Japan
Scott Shibasaki
Voice: +81 3 5225 6614
Fax: +81 3 5229 7253
s_shiba@optronics.co.jp
Reprint Services
Voice: +1 (413) 499-0514
Fax: +1 (413) 442-3180
editorial@photonics.com
Mailing addresses:
Send all contracts, insertion orders
and advertising copy to:
Laurin Publishing
PO Box 4949
Pittsfield, MA 01202-4949
Street address:
Laurin Publishing
Berkshire Common, 2 South St.
Pittsfield, MA 01201
Voice: +1 (413) 499-0514
Fax: +1 (413) 442-3180
E-mail: advertising@photonics.com
89 North .....................................................................................................................................................47
www.89north.com
a
Andor Technology ..............................................................................................................................41, 42
www.andor.com
Applied Scientific Instrumentation ...........................................................................................................6
www.asiimaging.com
b
Blue Sky Research ................... .................................................................................................................19
www.blueskyresearch.com
c
Cargille Laboratories ............... ...................................................................................................................8
www.cargille.com
Cooke Corporation Ltd. ........... ..............................................................................................................CV3
www.cookecorp.com
CVI Melles Griot ...........................................................................................................................15, 17, 27
www.cvimellesgriot.com
e
Edmund Optics ........................ .................................................................................................................20
www.edmundoptics.com
f
Finger Lakes Instrumentation ..................................................................................................................46
www.flicamera.com
g
Gooch & Housego ................... .................................................................................................................45
www.goochandhousego.com
h
Hamamatsu .............................. ...................................................................................................................8
www.hamamatsucameras.com
i
Incom Inc. ................................. ..............................................................................................................CV2
www.incomusa.com
Iridian Spectral Technologies ....................................................................................................................9
www.iridian.ca
l
Lumen Dynamics Group Inc. ................................................................................................................CV4
www.ldgi.com
Lumencor Inc. .......................... .................................................................................................................13
www.lumencor.com
m
Mad City Labs .......................... .................................................................................................................22
www.madcitylabs.com
Mightex Systems ..................... .................................................................................................................18
www.mightex.com
n
Newport Corp. .......................... ...................................................................................................................3
www.newport.com
o
Optical Building Blocks Corp. ....................................................................................................................5
www.obb1.com
Optical Society of America ..... .................................................................................................................42
www.osa.org/biomedicaloptics
p
PI (Physik Instrumente) L.P. .....................................................................................................................34
www.pi.ws
Picoquant GmbH ..................... .................................................................................................................23
www.picoquant.com
Prior Scientific Inc. .................. .................................................................................................................37
www.prior.com
r
Raptor Photonics Ltd. ............. .................................................................................................................18
www.raptorphotonics.com
s
Sutter Instrument .................... .................................................................................................................16
www.sutter.com
Sydor Optics Inc. ..................... .................................................................................................................16
www.sydor.com
ND11_Ad Index_Layout 1 10/27/11 11:34 AM Page 49
POSTSCRIPTS
T
he immunological battle against
AIDS could someday be won in
our genes with a little help from
a few glow-in-the-dark cats.
A new genome-based immunization
strategy from the Mayo Clinic in
Rochester, Minn., could help fight feline
AIDS and even illuminate ways to combat
human diseases, including HIV/AIDS,
which has killed more than 30 million
people. There is no effective vaccine
on the horizon.
Each year, millions of cats suffer
and die from feline immunodefi-
ciency virus (FIV), which causes
AIDS in cats the same way HIV
does in people: by depleting the
bodys infection-fighting T cells.
Naturally produced protective
proteins called restriction factors
are ineffective against FIV in cats
and HIV in humans.
The interdisciplinary research
team looked to jump-start evolution
by creating cats with an inborn im-
munity to the virus. Normally, over
vast spans of time, restriction factors
evolve to defend mammals against
virus invasion, but the Mayo physi-
cians, virologists, veterinarians and
gene therapy researchers, working
with scientists in Japan, didnt want
to wait that long.
They devised a way to insert a
rhesus monkey restriction factor
known to block FIV infection in a
culture dish into the cat genome,
creating cats that produce the pro-
teins on their own and even pass the
genes on to their kittens. A jellyfish
gene, added for tracking purposes,
has the side effect of making the
offspring cats glow green.
The technique, called gamete-targeted
lentiviral transgenesis, involves inserting
genes into feline eggs before they are fer-
tilized. This is the first time it has been
successful in a carnivore. The method is
highly efficient, so that nearly all offspring
have the genes, and the defense proteins
are made throughout each cats body.
The findings appeared online in Nature
Methods.
This specific genome modification ap-
proach will not be used directly to treat
people with HIV or cats with FIV, but
it will help medical and veterinary re-
searchers understand how restriction
factors can advance gene therapy for
AIDS in both humans and cats.
One of the best things about this bio-
medical research is that it is aimed at
benefiting both human and feline health,
said Dr. Eric Poeschla, a molecular biolo-
gist at the Mayo Clinic and leader of the
study. It can help cats as much as peo-
ple.
GENETIC APPROACH
shows bright promise against AIDS
A genome-based immunization technique could shed light
on ways to fight feline immunodeficiency virus and even
HIV/AIDS, among other diseases. A jellyfish gene included
for tracking purposes makes the offspring cats glow green.
Courtesy of Mayo Clinic.
Laura S. Marshall
laura.marshall@photonics.com
ND11_Postscripts_Layout 1 10/27/11 10:29 AM Page 50
high resolution
5.5 megap|xe|
readout noise
< 1.4 e|ectrons
dynamic range
> 22 000 : 1
maximum frame rate
100 frames / s
pcc.edge - the first cemere system
with the revcIuticnery sCNOS imege senscr
Bringing to light! The new camera system pco.edge represents a perfect
combination of high resolution, extremely low read out noise, and superior
dynamic at low light, for excellent image quality even at high frame rates.
Discover the new possibilities in the range of high performance applications.
For more information please visit
www.cookecorp.com/scmos-cameras/pcoedge/
www.cookecorp.com
In Europe. www.pco.de
nd11_CookeCorp_PgCVR3_Layout 1 10/27/11 10:31 AM Page CVR3
nd11_LumenDynamics_PgCVR4_Layout 1 10/27/11 10:32 AM Page CVR4

Biophotonics201111 DL

Загружено:

Сведения о документе

Исходное описание:

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Biophotonics201111 DL

Загружено:

Авторское право:

Доступные форматы

November/December 2011

Search Biophotonics-Related News

nd11Online_Layout 1 10/27/11 10:12 AM Page 9

ND11_Breakthrough Leads_Layout 1 10/27/11 11:48 AM Page 43

and operation at >385 fps with binning. Be-

Вам также может понравиться