Analysis: Manual Colorview

analySIS
Manual
ColorView
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ColorView II
ColorView II - Camera - technical specifica-

tions 1
ColorView II
Handbook Layout 2
Camera and light microscope 3
WHITE BALANCE .....................................................................3

Monitor Settings for White Balance ................................4
Executing White Balance ...............................................4
Online shading correction 5
OPERATION OF THE SHADING CORRECTION ....................6

ACQUIRING CORRECTION IMAGES ......................................7
Enabling Shading Correction ...............................................9
Repeat or Update the Reference Acquisitions .....................9
USE OF THE ONLINE SHADING CORRECTION ..................10
TECHNICAL NOTE .................................................................10
intX and Online Shading Correction ..................................10
Image Acquisition 11
ECVII0804
ACQUISITION METHODS ......................................................11

Live-Acquisition ......................................................11
Snapshot ................................................................11
intelligent Exposure ................................................11
ACQUISITION .........................................................................11
CAMERA CONTROL ..............................................................12
Contents
Exposure Time ....................................................... 12

White balance ........................................................ 13
Color Settings ........................................................ 13
Sharpen filter .......................................................... 14
Automatic Gain Display and Scaling ...................... 15
Black balance ......................................................... 16
Sharpness Monitor ................................................. 17
Partial Readout ...................................................... 18
SNAPSHOT ............................................................................ 19
INTELLIGENT EXPOSURE .................................................... 19
NetCam 23
INSTALLING THE WEB SERVER .......................................... 23

NETCAM IN THE INTRANET ................................................. 24
NETCAM IN THE INTERNET ................................................. 24
USING NETCAM ..................................................................... 25
Usage in Black & White mode 27
Gamma Correction ............................................................ 28

Display and usage of the acquisition with Real time FFT .. 28
Camera Set Up 29
SOFTWARE CAMERA CONTROL ......................................... 30

The Input tab ................................................................ 31
The Display tab ............................................................ 34
The Format tab ............................................................ 38
Contents
Appendix I: Microscope Settings 40
Appendix II: Camera Installation 43
INSTALLATION PROCEDURE - WINDOWS 2000 ................ 45

INSTALLATION PROCEDURE - WINDOWS XP ................... 58
WHEN ENCOUNTERING PROBLEMS WITH THE CAMERA... 66
CE DECLARATION OF CONFORMITY 68
ColorView II - Camera - technical specifications
ColorView II
ColorView II - Camera - technical specifications
Definition ColorView II is a quick, high resolution CCD color camera used for bio-
medical and material sciences applications. It was especially developed
for acquisitions using light microscopy, has a special compact design, and
can be connected to all types of light-microscopes via the C-mount. Its high
dynamic scope enables it to acquire images which have very light and very
1
dark areas
ColorView II offers the following technical specifications:
ColorView II
Dynamic range 3 x 12 bit using color mode
12 bit using black & white mode
Exposure times 100 µs to 160 s
Frame rates 25 / s in Search Mode
14 / s in Focus mode
5 / s in high resolution mode
Resolution 2080 x 1544 in color mode
1040 x 772 Pixels in black-white mode
Binning 2x, 3x
Cooling Peltier-cooling: 10°C @ 25° ambient
temperature
Camera Mount Standard C-Mount
PC-interface FirewireTM (IEEE 1394)
ColorView II • Handbook Layout
System Requirements To properly use ColorView II, the following system requirements for your
computer and your analySIS image analysis system should be taken into
consideration:
Hard- / Software Required
CPU P4 with at least 1,8 GHz
Memory 256 MB
Hard Disk Storage 20 GB
Interface FirewireTM (IEEE 1394)
Operating system Windows 2000 Professional,
Windows XP Pro
analySIS analySIS 3.2
Build 625 or higher
Modern cameras with the respective software offer numerous possibilities

for correcting and modifying the acquired image. They enable the image to
be optimized quickly. However, the entire system, the sample, the micro-
scope with all its possibilities, and the camera all must be optimally set in or-
der to attain the best possible results.
Handbook Layout
This handbook will help you use your system to optimally acquire images.
Camera Adjustments All basic camera settings are handled by the software via one or more logical
input channels. You will find the input channel options for color and black &
white use in the chapter entitled "Adjusting the Camera". This chapter is at
the end of the book, since hardly any alterations are made to the input chan-
nels in day-to-day use.
Camera and light mi- The chapter entitled "Camera and Light Microscope" portrays the elements
croscope of the microscope which have the greatest influence on the image quality.
The correct microscope settings are explained.
Image Acquisition The chapter entitled "Image Acquisition" deals with the uses and possibili-
ties of a digital camera, as well as the software required for image acquisi-
tion. This chapter will be the most used chapter for day-to-day use. The in-
2
dividual acquisition methods (live-acquisition, single image, intelligent expo-

sure) are explained in this chapter in detail.
Usage in Black & White In addition to the normal color mode, ColorView II also offers a black & white
mode mode. The chapter entitled "Usage in Black & White mode" describes the
setup of this mode as well as the alterations in camera usage.
Related Topics
Camera Set Up 29
Camera and light microscope 3
Image Acquisition 11
Camera and light microscope
Camera and light microscope

Both the camera settings as well as the microscope settings are vital for the
quality of the image to be acquired. Only the most important rules for the mi-
croscope settings are mentioned here; a more thorough explanation to the
topic can be found in the appendix:
1) The illumination must be parallel.
2) Use ND-filters to reduce the illumination intensity; do not alter the lamp
voltage.
3) Establish Köhler illumination conditions.
4) Select the appropriate total magnification and the objective for the
problem in question.
White Balance
The human eye reacts very sensitively to color variations which are per-
ceived as a color tinging effect. Therefore, when wanting to acquire quality
images with a digital camera, it is vital that the individual color channel set-
tings are set so that the entire image does not show color tinging. The ad-
justment of the color channels and the white balance depend on the illumi-
nation conditions. When changing the lamp or altering the lamp voltage, the
white balance should be repeated afterwards accordingly.
When using white balance, the individual color channels are scaled in a way
that the white or neutral gray area of the image displayed on the monitor is
displayed correctly as white or gray. The white balance can be carried out
automatically on each image or manually for individual images.
Mode of operation An area which is, for the most part, uniformly gray or white is selected on the
image. Such an area must contain an equal intensity of the three color com-
ponents : red, green and blue. White balance sets the color channels in such
a way that the requirements for the selected area are satisfied as much as
possible. To be able to carry out an effective white balance, you need the
respective white or gray area.
White balance is carried out on a part of the image (ROI). The program alters
the color settings in such a way that the image area in the ROI appears as
3
white or gray as possible. After you have once carried out the white balance,
you can reapply it any time to a running live-image acquisition.
Standardized White Bal- A test image is acquired with the camera using optimal illumination for the
ance standardized white balance. The individual color channels are now set so
that the acquisition also appears white or gray on the monitor. These chan-
nel settings are saved and used automatically for all additional acquisitions.
Manual White Balance A manual white balance is carried out on the current single image / snap-
shot. To do this, you need an area in the image which is for the most part
gray or white.
Microscope Settings Use the same microscope settings for the white balance that you will use for
the acquisitions later. This especially holds true for the filter and the lamp
voltage settings. The lamp voltage of a 12V halogen lamp should be set to
9V. It should, however, not be set lower than 5V. Use the color neutral ND-
filter, should you have to reduce light intensity.
Related Topics
Appendix I: Microscope Settings 40
ColorView II • White Balance
Monitor Settings for White Balance

Monitor Settings The monitor settings are decisive for the image you see on the monitor. Set
your monitor to the appropriate color temperature, should your device allow
this. To do this, compare the image's color impression on the monitor and in
the eyepiece.
Note The color temperature setting can be altered on most monitors via the mon-
itor menu buttons located on the monitor. In other cases, the color temper-
ature can be altered via the operating system.
How to... This is how you change the color temperature using the operating system:
1) Right click on Desktop.
" The Desktop's context menu is opened.
2) Select the Properties entry from the context menu.
" The Display Properties dialog box is opened.
3) Select the Settings tab.
4) Click the Advanced... button.
" Another Display Properties dialog box is opened. The appearance
and function of this dialog box depends on the graphics card in-
stalled on your system and your monitor.
" Search for a tab with the name Color or Color Management.
Note The appearance of the operating elements might differ from the one being
described here because the adaptation of the color temperature is not part
of the operating system, but rather is made accessible by the monitor man-
ufacturer or the graphics card being used.
5) Select a color temperature whose color impression best corresponds
to that seen in the eyepiece.
" Should your system offer a slide control with the options warm and
cold, do not alter your color temperature with these.
Executing White Balance

Specialized microscopy It is very difficult to execute a white balance in some microscopic methods,
techniques i.e. DIC or polarization. In these cases, execute the white balance using the
bright field method and only then begin with the special methods.
4
Transmitted Light When using the transmitted light method, look for a position through which
the light passes through the cover slip, embedding material, and microscope
slide, but not through the specimen. Should there not be such an area, re-
move the object to be observed. Thus, the lamp light used for illuminating
the specimen falls directly on the objective. This is white when using a white
lamp.
Reflected light When using the reflected light method, you need the appropriate reference
surface. Standardized gray cards are best for this. Should one not be avail-
able, you can also use white laboratory porcelain or neutral ND-filter paper.
Should your samples contain very reflective areas, defocus your microscope
when setting the white balance.
How to... This is how you set up the white balance:
1) Acquire an appropriate snapshot.
2) Use the Image > Camera Control... command.
" The Camera Control dialog box is opened.
Related Topics
Camera Control 12
Online shading correction
3) Click the Set ROI for White Balance button.

4) Pull the ROI's red frame shown in the image document to a position
which is more or less a uniform white or gray.
" Change the size of the area by keeping the mouse depressed and
moving the mouse. The ROI used for the white balance cannot be
made to any large size possible.
" The ROI can be difficult to see if the image document has a reddish
shade. In this case, use the Special > Preferences > View com-
mand to change the color of the frame. You can select a more ap-
propriate color from the Primary list located in the Mouse cursor
group.
5) Rightclick to confirm position and size of the ROI.
6) Click the White Balance button to conduct an automatic white balance.
" The settings for the individual color channels are optimized in the
set area. The result is directly visible in the image buffer's image.
7) Click the Color Settings button.
" The Color Settings dialog box is opened.
8) Click the Define...button.
" The Define Default Color Settings dialog box is opened.
9) Click the Current settings button to adopt the current values for the rest
of the work.
" When using this setting, the white balance is valid for all images
which are acquired with this input channel during the current ses-
sion.
10) Click the OK button to return to the Color Settings dialog box.
11) Click the Standard button to save these values as general standards.
" The current values for the white balance are now saved and will be
imported when the program is restarted. A manual change of the
color settings changes the white balance.
The program alters the color settings in such a way that the image area in
the ROI appears as white or gray as possible. After you have carried out the
white balance once, you can reapply it any time to a running live-image ac-
quisition.
Online shading correction 5

Optical systems consisting of camera and microscope generate image inho-
mogeneity, or so called shading, even if care was taken with setting up the
devices.
The left illustration shows an
image where the online
shading correction has not
been activated. In compari-
son to the image on the
right, one can notice the in-
homogoneities in the illumi-
nation.
ColorView II • Operation of the Shading Correction
Shading Correction For a shading correction, the image errors are first determined in order to be
able to make corrections to the acquired image. Using the online shading
correction of ColorView II, these corrections already take place in the live-
image.
Operation of the Shading Correction

The correction of the image proceeds pixel to pixel with the aid of two differ-
ent reference image series: one consists of dark images for each resolution
of the camera, and the other consists of correction images. Before the online
shading correction is able to be used, you must acquire these reference im-
ages.
The illustration explains the
principle of the online shad-
ing correction.
Dark image and correction
image are transferred from
the PC to the camera based
on the magnification and ob-
servation method.
The corrections based on
these images are done in
the camera.
The image transferred to the
PC has already undergone
shading correction.
6
Dark Images A dark image is an acquisition in which no light falls on the camera. Errors
can occur from noise or defective pixels which can be corrected by the dark
image. An own dark image is required for each resolution of the camera.
Correction Images The illumination of the entire optical system without the object (with refer-
ence samples in reflected light mode) is shown. In addition to the camera
characteristics, the microscope's optical characteristics, especially the ob-
jective being used contribute to the correction image. A correction image for
each camera resolution and objective combination must be made accord-
ingly.
Acquisition Methods The utilized acquisition method also plays a vital role in addition to the al-
ready mentioned factors of the actual cameras and objectives. The online
shading correction has been aligned with transmitted light, reflected light,
DIC and Phase Contrast microscopy. When acquiring the correction imag-
Acquiring Correction Images
es, you will be asked for the set methods of acquisition. Should you want to
employ various methods, you must repeat the acquisition of the correction
image for each method used.
Note When using a magnification changer, correction images must be acquired
for each possible total magnification.
Acquiring Correction Images

Please be very careful when acquiring correction images for the online
shading correction, since false correction acquisitions can lead to artifacts in
the images. The actual acquisition process is controlled by a wizard. This
wizard provides you with a step-by-step procedure.
Requesting Reference A correction acquisition is always linked to a concrete magnification; a XY-
Acquisitions calibration of the objective being used must be carried out before the acqui-
sition of the correction images.
Within the scope of the preparation of the acquisition, the wizard asks you if
the correction acquisition is to be made only for the current magnification or
for all calibrated magnifications.
Transmitted Light 1) Disable the preprocessing and postprocessing of the image acquisi-
Mode tion.
" To do this, clear both Enabled check boxes in the Macro tab of the
Configure Input dialog box (command Image > Configure Input...).
2) When using a tungsten-halogen lamp, set the same lamp voltage as
when using normal operation.
" The lamp voltage should not be below 5V. Use ND-Filters to reduce
possible brightness.
Note ND-Filters belong to the basic components of the microscope. They will be
put into the light path to reduce brightness without altering the color temper-
ature. Should your microscope not have a ND-filter or not be able to have
one installed, you must instead reduce the lamp voltage.
3) Set the illumination method to brightfield. This also holds true if you
want to use special methods at a later time.
4) Bring the objective with the smallest magnification, by which the con-
7
densor front lens still has to be used, into the light path and set the
Köhler illumination.
5) Set the camera's exposure time to a value of > 50ms by lowest resolu-
tion.
" The value in the online histogram should be around 2,000.
6) Bring the object into the light path and bring it into focus.
7) Defocus by increasing the space between the objective and the object
until the object is recognizable as a whole but the details of the struc-
ture are no longer visible.
8) Move to a position on the slide where neither the sample is visible, nor
are there any scratches or contaminations.
" Should such a position not be able to be found, remove the micro-
scope slide from the microscope and acquire the reference images
without the object.
9) Begin the acquisition of the reference images with the Image > Acquire
Reference Images... command.
" The Online Shading-correction dialog box appears.
Related Topics
Köhler Illumination 41
ColorView II • Acquiring Correction Images
10) Follow the wizard's instructions to acquire the reference images.

Reflected Light Mode 1) Disable the preprocessing and postprocessing of the image acquisi-
tion.
" To do this, disable both Enabled check boxes in the Macro tab of
the Configure Input dialog box (command Image > Configure In-
put...).
2) When using a tungsten-halogen lamp, set the same lamp voltage as
when using normal operation.
" The lamp voltage should not be below 5V. Use ND-Filters to reduce
possible brightness.
Note ND-Filters belong to the basic components of the microscope. They will be
put into the light path to reduce brightness without altering the color temper-
ature. Should your microscope not have a ND-filter, please contact your
supplier.
3) Set the illumination method to brightfield. This also holds true if you ad-
ditionally want to use special methods at a later time.
4) Bring the objective with the smallest magnification, by which the con-
densor front lens still has to be used, into the light path and set the
Köhler illumination.
5) Set the camera's exposure time to a value of > 50ms by lowest resolu-
tion.
" Your online histogram's gray value should be around 2,000.
6) Bring the reference sample into the light path and set the Köhler illumi-
nation.
" The reference sample must be color neutral and have a flat surface
devoid of defects or optical structures visible through the micro-
scope.
Note Should you not have access to a reference sample, you can also use a piece
of opaque glass. Should you want to use another sample, please be sure to
defocus it.
7) Increase the distance between the objective and the reference sample
until the surface structures of the reference sample are no longer visi-
ble.
8
Warning Please note that the focus depth is high when using objectives with lower
magnification. In this case, you have to defocus more than with objectives
with a magnification of 10x or higher.
When using reference samples with transparent surfaces, it is important to
be sure that a section within the reference sample is, by all means, not
shown.
8) Begin the acquisition of the reference images with the Image > Acquire
Reference Images... command.
" The Online Shading-correction dialog box appears.
9) Follow the wizard's instructions to acquire the reference images.
10) Carry out a new white balance when first using the online shading cor-
rection.
Related Topics
Köhler Illumination 41
Enabling Shading Correction
Enabling Shading Correction

When finishing the reference acquisitions, you will be asked if you want to
activate the online shading correction. Independent of the decision as of
now, you can activate or deactivate the online shading correction with the
Image > Configure Input > Input command any time. The settings of the on-
line shading correction remain even after the program has been restarted.
Repeat or Update the Reference Acquisitions

The online shading correction functions optimally as long as the illumination
settings are not changed.
Sample acquisitions Sample acquisitions enable you to check the quality of the online shading
correction. To do this, acquire an image of a sample with a background as
even as possible. Now, measure the acquired sample's intensity distribution
with the commands in the Measure >Intensity Profile submenu. Apart from
the sample's signature, the measured profile is even if the online shading
correction is perfectly set.
Example for an intensity
profile without shading cor-
rection (or poorly executed)
and with shading correction.
The intensity profile shows a
clear decline along the X-ax-
is.
Alterations to the illumi- If you switch to another illumination method supported by the online shading
nation method correction, you must make a new correction acquisition if none exists for the
method. Should the required acquisitions already exist, then you can set the
appropriate illumination method with the Image > Configure Input > Input
command. To do this, select the appropriate entry from the Illumination
method list.
Adding/changing objec- If you add new objectives or change already installed objectives, you must
tives acquire correction images for the new objective.
ColorView II • Use of the Online Shading Correction
Altering the lamp voltage Smaller alterations made to the lamp voltage do not influence the online
shading correction. Larger alterations cannot be balanced by the online
shading correction. Therefore, use the same lamp voltage you utilize in your
normal routine use for acquiring the correction images. Use ND-filters to re-
duce the brightness or acquire new reference images.
Use of the Online Shading Correction

Please note the following instructions for the use of the online shading cor-
rection in order to be able to make optimal acquisitions.
Before acquisition Before the acquisition procedure, make sure that the optical system is clean.
Check the lamp voltage and check if the Köhler illumination has been set. In
addition, no alterations should have been made to the light path on the mi-
croscope.
Setting magnification The correct magnification must be set for the use of the online shading cor-
rection. Use the SetMagnification$(); command, but not in the input channel
because in this application it is executed too late.
Automatic microscope It is not necessary to set the magnification when using an automatic micro-
scope which is remote controlled by the application.
Technical Note
Organizing the reference Dark images and correction images are saved to the hard disk. In order to
images be able to work with a correct online shading correction when having
changed the camera, the application creates an individual directory for sav-
ing the reference images. This directory is in the application's working direc-
tory. Its name begins with DGSISFW and continues to contain information
about the serial number of the camera being used. This directory should not
be moved or renamed.
The reference images file names are also coded in such a way that the on-
line shading is able to automatically access it. Should one or more of the ref-
erence images have been deleted, the application immediately tells you that
a missing image is required and automatically requires you to reacquire the
missing reference image.
10
File Format The reference images are saved as normal TIF images.
Requirements The filter in front of the camera's CCD chip and all of the microscope's opti-
cal parts must be clean. Otherwise, dust particles or fingerprints would ap-
pear in the correction images. The shading correction can of course correct
such contaminations, as long as they remain unchanged. Should a contam-
inant be moved or removed, the shading correction would give you a ghostly
image of the former contamination in each new image. Check the filter be-
fore acquiring the reference images and clean it with a LensPen, if neces-
sary.
intX and Online Shading Correction

The intelligent Exposure acquisition method works problem-free together
with the online shading correction. In some cases, however, the shading
correction can result in over exposure. You will then receive an error mes-
sage when executing a white balance.
Adapting intelligent Ex- In this case, click the Options...button located in the intelligent Exposure di-
posure alog box. Reduce the Dynamic limit in %value in the intelligent Exposure
Options dialog box to approximately 70.
Image Acquisition
Image Acquisition
The chapter about "image acquisitions" deals with the various acquisition
methods you can use with the image-analysis software.
Acquisition Methods
The software offers you various acquisition methods for various tasks.
Live-Acquisition
When using live acquisition, the image is continually transferred from the
camera to the monitor. This mode is suited for setting various parameters,
including image segments, focussing, and camera settings. The live acqui-
sition is concluded after a snapshot has been acquired. You can use the Im-
age > Camera Control... command to do numerous camera settings during
a live acquisition.
Snapshot
The Image > Snapshot command concludes the live acquisition and saves
the current image to the active image buffer. The same settings which are
used for the live acquisition are also used for the snapshot. You can make
a snapshot without acquiring a prior live acquisition.
intelligent Exposure
The Image > intelligent Exposure... command offers you an extensively au-
tomated image acquisition. The automated functions being used coordinate
the amplification of the image signals and the exposure time of the acquired
image so that a good image result is guaranteed. The exposure time of a live
acquisition, through the resetting of the camera's acquisition mode, are kept
so short that a very good adjustability during the live-image is guaranteed.
Acquisition
11
Use the Image > Acquire command to display the acquired image on the monitor live.
You may also You may also click the Standard (button bar) > Acquire button.
Available The command is not available if the active image buffer is write protected.
What will happen... The live-image is displayed in the active viewport. The camera symbol locat-
ed in the status bar blinks and the name of the camera appears in red. As
long as the live-image is shown in the active viewport, the image-analysis
software only offers commands which can be effectively used.
Why use live-images? You need the live-image for focussing, or when arranging the camera or the
sample. The Image > Snapshot command stops the live-image display and
takes the image last shown in the active image buffer.
Related Topics
Acquisition 11
Snapshot 19
intelligent Exposure 19
ColorView II • Camera Control
live-image settings You can determine the settings for the image acquisition in the logical input.
Use the Image > Set Input...and Image > Configure Input... command to cre-
ate an input and to determine its properties.
Editing other images in The live-image is shown in only one viewport. If you are using two monitors
live mode or have numerous viewports in your image window, you can enable another
viewport in live mode and at the same time load another image and display
it in this viewport, for example. In addition, you can measure another image
with the interactive measurement functions, save them, or insert them into a
database while live mode is active.
Live overlay The Live overlay function enables you to write and draw directly onto the im-
age's overlay during the live acquisition. To do this, use the buttons in Over-
lay button bar.
Available The live overlay is only available if you have activated it with the Image >
Configure Input command located on the Display tab.
Camera Control
Use the Image > Camera Control... command to alter the settings for acquisition with the camera.
The Camera Control command and the appropriate dialog box enables you
to attain control over numerous setting possibilities required for acquiring
single images and live acquisitions. The commands in this dialog box are
closely interlocked with the settings available from the Image > Set Input
command.
Available The Camera Control command is only available if the Set Input dialog box
is closed.
You may also You can also click the Standard (button bar) > Camera Control button.
Color operation and The appearance and the functions of the Camera Control dialog box depend
black & white operation on whether or not color or black & white images are taken. This section de-
scribes the camera controls for the camera's color mode; a description for
the black & white operation can be found in the section entitled "Operation
12
in Black and White Mode".

The Camera Control dialog
box for a ColorView II cam-
era using color operation
Exposure Time
The exposure time required for high-quality acquisitions depends on the il-
lumination, the properties of the sample, and the camera being used. First,
remove possible ND-filters to be able to manage with lower exposure times.
You can also additionally set the prism or the phototube's mirror to "only
Related Topics
Live overlay 36
Camera Control
photo". Should the result not be adequate enough, you can then increase
the lamp voltage. After you have increased the lamp voltage, it is recom-
mended that you redo the white balance.
Exposure Time You can set the camera's exposure time in the Exposure time group.
Click the - and + buttons to alter the exposure time in pseudo-logarithmic
steps. You can also directly change the exposure time in the corresponding
text field or manipulate it using the arrow buttons next to the text field. The
minimum and maximum exposure times depend on the camera being used.
Auto Click the Auto button to enable the automatic exposure time settings. The
Auto button is only available in live-mode.
White balance
The white balance proceeds after the description shown above (S. 4).
Color Settings
Click the Color Settings button to manually alter the camera's color settings.
The current settings can be altered using the five slide controls: Red, Green,
Blue, Gamma, and Saturation.
Red, Green, Blue The Red, Green and Blue slide controls can each be set from 0,00 to 20,0.
The set value is the factor with which the signal of the corresponding color
channel is multiplied.
13
Gamma Use the Gamma slide control to obtain a nonlinear contrast enhancement.
Please note that this gamma filter does not only affect monitor display but
also alters actual image information. If you are planning on quantitatively
evaluating your images at a later time, you are advised to use the value 1.0
for the gamma filter. The Oper menu provides you with commands for ad-
justing image contrast at any later time.
What is gamma? As is the case with the slide controls of the individual color channels, the nu-
merical value for gamma effects how the illumination intensity of the pixels
of the camera CCD chip is converted into the color value for the correspond-
ing pixel in the image buffer. If gamma equals 1, the conversion from illumi-
nance into gray/color value is linear. A nonlinear contrast enhancement
results should the gamma not be equal to 1: if gamma is smaller than 1, the
darker gray values will be spread more significantly than the brighter ones.
The result will be enhanced contrast in the lighter areas of the image. The
image as a whole will become darker. If gamma is greater than 1, the darker
Related Topics
White balance 13
gray values will be spread more significantly than the brighter ones. The re-
sult will be enhanced contrast in the dark image areas. The image as a
whole will become lighter.
Saturation The Saturation slide control enables you to alter the color rendering from a
black and white signal to maximum color saturation. In most cases, the
"good" setting of this value is seldom higher than the range of 0,10 to 0,20.
The Gamma slide control's value range is from 0,00 to 2,00; the Saturation
slide control's range is from 0,00 to 0,99.
Default Click the Default button to use the default values defined in the Define De-
fault Color Settings dialog box from now on.
Define… Click the Define... button to set the standard color settings. The Define De-
fault Color Settings dialog box is opened.
Factory settings Click the Factory settings button to return the color settings to their original
levels. The factory settings have been adapted in such a way that the color
rendering with a specific equipment configuration is ideal for a large scope
without any additional changes having to be made.
Current Settings Click the Current settings button to make the current values be the future
standard.
Sharpen filter
Use the sharpen filter to increase the sharpness using edge enhancement
14
already with the live acquisition.

Sharpen Filter Click the Sharpen Filter On/Off button to activate/deactivate the sharpen fil-
On/Off ter. The manner in which the acquired image is affected by the sharpen filter
depends on the sharpen filter parameter. You can adjust this parameter by
using the Sharpen Filter Settings dialog box.
Clicking the Sharpen Filter On/Off button produces the same results as se-
lecting/clearing the Activate check box in the Sharpen filter group in the In-
put tab. The button is simply a more convenient alternative.
For more information about the sharpen filter and sharpen filter parameter
have a look at the chapter "The Input tab S. 31"
Sharpen Filter Click on the Sharpen Filter Settings button to open a dialog box where you
Settings can set the sharpen filter parameter:
Camera Control
Available The Sharpen Filter Settings button is only available if the sharpen filter has
been activated.
To adjust the numerical value of the sharpen filter parameter, use the mouse
to move the slide control to the right or left. This is a convenient alternative
to using the Input tab - where you can also adjust this numerical value (in
the Sharpen field).
Automatic Gain Display and Scaling

ColorView II uses a dynamic range of 12 bits in black and white mode or 3
x 12 bits in color mode. However, only 8 bits, respectively 3 x 8 bits can be
used for the display and additional editing of images. Therefore, the data
transferred from the camera to the computer must be adjusted directly dur-
ing acquisition so that this condition is fulfilled. There are different methods
for this purpose which can be utilized based on the specimen being ob-
served as well as the microscope and camera settings.
No Settings The four lowest bits are ignored should you not make any settings. Thus, at
large, only imperceptible color alterations to individual pixels occur.
Fixed scaling The "Fixed scaling" method gives you a fixed defined intensity area to be
used for each channel. To do this, set the limits in the range from 0-4095. All
signals which are outside the defined areas will be ignored. It is not possible
to set a bottom limit with color cameras due to the fact that this would lead
to strong color alterations. Use this method with images with weak contrasts
or to select a specific signal area (fluorescent microscopy).
Automatic gain display The "Automatic gain display" method extracts the area to be used based on
the actual signals on hand. The strongest of these signals will be ignored.
15
Use this method with images with a high dynamics range or low maximum
signal strength.
Use automatic Click the Use automatic gain display button to activate the automatic gain
gain display display. The live-image, independent from the illumination conditions, is al-
ways shown in optimal contrast. The behavior of the "automatic gain dis-
play" depends on whether or not this function's desired overflow has been
set and on which area of the image the contrast enhancement is going to
take place. The desired overflow can be set using the Image > Configure In-
put command located on the Display tab. The following possibilities for se-
lecting the image area can be found in the Camera Control dialog box:
Histogram cal- Click the Histogram calculation on full image button to carry out the contrast
culation on full optimization on the basis of the entire image. This option is recommended if
image the structures to be observed are equally dispersed throughout the entire
image.
Histogram cal- Click the Histogram calculation on crosshair button to only consider the pix-
culation on els along a strip which is only a few pixels wide in the horizontal and vertical
crosshair direction.
Histogram cal- Click the Histogram calculation on full image button to carry out the contrast
culation on ROI optimization on the basis of a rectangular area within the image. You can
use this option if relevant details are visible in a part of the image. The dis-
play can thus be optimized for this area.
Set ROI for his- Click the Set ROI for histogram button to interactively set the rectangular
togram area of the image for optimizing the contrast. Pull the ROI's frame shown in
the image document to the desired position and set its size by moving the
mouse and keeping the right mouse button depressed. Rightclick to confirm
position and size of the ROI.
Use fixed scal- Click the Use fixed scaling button to use a fixed scaling amplification. In con-
ing trast to the automatic gain display, the fixed scaling utilizes a predefined sig-
nal range which is manipulated so that it uses the entire dynamic area.
When using the fixed scaling, the image is no longer displayed optimally if
the illumination or sample are altered.
Automatically Click the Automatically adjust fixed scaling button to automatically set the
adjust fixed scaling on the current conditions. This button is only available during an ac-
scaling tive live acquisition.
Manually ad- Click the Manually adjust fixed scaling button to manually set the upper right
just fixed scal- hand limit for the fixed scaling. The Fixed scaling dialog box is opened.
ing
Move the Right limit slide control or directly enter the desired numerical val-
ue into the text box.
Black balance
The black balance is used for background correction. The program calcu-
lates the mean value for each color channel in an area (ROI) which you de-
fined. These values are subtracted from all pixels in the entire image. The
16
black balance is to be used predominately for fluorescence acquisitions in

order to balance the undesired background in the acquisition. A discolora-
tion of the images occurs if the black balance is used with brightfield acqui-
sitions.
Functionality The black balance is carried out based on the rectangular area in the image,
in which a uniformly dark-gray to black signal is to be located. The mean sig-
nal value is determined for each color channel within this area. This value is
reduced from each pixel when applying the black balance.
How to... This is how you carry out a black balance:
1) Take a snapshot which contains a suitable area to be used for black
balance.
2) Use the Camera Control command.
3) Click the Set ROI for Black Balance button to adduct an area of the im-
age for the black balance.
4) Pull the ROI's red frame shown in the image document to a position
which contains an area which is as uniformly dark as possible.
Camera Control
" Change the size of the area by keeping the mouse depressed and
moving the mouse. The ROI used for the black balance cannot be
made to any large size possible.
5) Rightclick to confirm position and size of the ROI.
" The values are calculated for the black balance on the area defined
in such a way.
6) Click the Black Balance On/Off button to activate the black balance.
" The correction values calculated for the black balance are deduct-
ed from all the pixels in the entire image.
The black balance function is available for live acquisitions as well as for the
acquired snapshots.
Sharpness Monitor
The Sharpness Monitor allows you to control the focus settings during a live
acquisition. You are also able to define a rectangular area of the image
which is to be analyzed for the sharpness monitor in the same way as for the
white balance and black balance. The maximum contrast serves as a mon-
itor for the sharpness in this area. Therefore this area should be limited to
an image segment in which the contrast is a good monitor for the sharpness.
Meaning of the Sharp- The Sharpness Monitor consists of a dialog box in which a relative measure-
ness Monitor ment of the sharpness is displayed by a changing bar which can be varied
between Blurred and Focused.
If you start a live acquisition with an activated Sharpness Monitor or activate

it during a live acquisition, it will show middle sharpness (50%) in each case.
This is because there is no absolute measurement for the sharpness in an
image and because the sharpness monitor only gives relative values which
are related to the initial situation which was present when starting the live
acquisition.
17
Focussing and Sharp- The length of the bar changes if you alter the focus settings during a live ac-
ness Monitor quisition in progress with an active sharpness monitor. In doing so, the
green markation bar shows the maximum sharpness reached since the live
acquisition was started. The bar shifts accordingly to the right should the
sharpness increase due to your changes. If the focus settings are already
optimal when starting acquisition, the bar can only shift in direction Blurred.
Reset to 90% Click the Reset to 90%button to set the sharpness monitor's relative value
to 90%. Utilize this function if you start working with a very blurry image and
if the sharpness monitor's bar reaches the limit to the right while focusing.
By resetting to 90%, the display gets more room for additional focussing.
The Reset to 90% function is also available if the sharpness drops very dras-
tically due to changes made to the microscope settings (i.e. changing objec-
tives) during an acquisition in progress.
How to... This is how you set the sharpness monitor for good acquisitions:
1) Set all other parameters (microscope settings, exposure time, white
balance) optimally. The application of the sharpness monitor should be
the last step before image acquisition.
2) Acquire a snapshot.
3) Use the Camera Control command.

4) Click the Set ROI for sharpness monitor button to interactively define a
rectangular area of the image for the sharpness monitor.
" To do this, select an area of the specimen for which a good sharp-
ness setting is especially important.
5) Begin a live acquisition.
" The Set ROI for sharpness monitor button becomes inactive.
6) Click the sharpness monitor on/off button to open the sharpness mon-
itor dialog box.
" The sharpness monitor begins with a value of 50%.
7) Vary the focus settings and observe the sharpness monitor while doing
so.
" The sharpness monitor bar shows the current sharpness, the green
markation shows the highest achieved sharpness.
" Mind the exposure time when setting the focus. The sharpness
monitor alters itself with a certain amount of delay when using larg-
er exposure times (> 100ms). Vary the focus settings in small inter-
vals and wait each time until the sharpness monitor has been
adapted as well.
8) Find the focus settings by which the sharpness monitor is at its maxi-
mum and use these settings for the acquisition.
Partial Readout
The partial readout is a method for fast live acquisition. A rectangular part of
the image is defined for the partial readout to which the readout is limited
during the live acquisition. A higher frame rate is attained due to the smaller
amount of data to be transferred.
Available The live overlay must be enabled for the partial readout. The live overlay can
be enabled via the Display tab located in the Configure Input dialog box.
Partial Readout and Clip- The partial readout and clipping functions both lead to smaller images than
ping those possible with the camera. The partial readout already works when the
image is read out of the camera; the image areas which are faded out by this
function are not transferred to the computer. The clipping, however, is then
18
calculated out of the complete image data in the computer; an acceleration

in the image acquisition is not intended. The frame rate, due to the very high
pixel count of the camera, using full resolution is often not enough to enable
good focussing during live-mode. In order to attain both high resolution and
high frame rates, you must do the following:
1) Start the live acquisition using full resolution.
2) Set the ROI for Partial Readout onto an important image detail.
3) Activate the Partial Readout.
4) Do the required settings.
5) Deactivate the Partial Readout.
" The image area set with the ROI for Partial Readout applies to the
following snapshot, should Partial Readout be enabled.
6) Acquire a snapshot.
Either a partial readout or a clipping is possible all the time. A partial readout
is disabled if you enable a clipping on the Format tab located in the Config-
ure Input dialog box. The ROI which was set for the partial readout remains
Related Topics
The Display tab 34
Snapshot
intact. Clipping is disabled if you enable a partial readout while clipping is en-
abled. The settings for the clipping, however, remain intact. They will be
once again utilized as soon as you disable the partial readout.
Partial Readout Click the Partial Readout On/Off button to activate the Partial Readout. In-
on/off stead of the camera's entire CCD chip, only a rectangular area which was
defined with the Set ROI for Partial Readout function is read. Clipping will be
disabled should it have been enabled. A predefined ROI is used in the image
center, should you activate the partial readout before a ROI has been set for
this function.
Set ROI for Par- Click the Set ROI for Partial Readout button to interactively set the rectan-
tial Readout gular area of the image to be read out. Pull the ROI's red frame shown in the
image document to a position on the image which is to be read out. Change
the size of the area by keeping the mouse button depressed and moving the
pointer. Rightclick to confirm position and size of the ROI. The Set ROI for
Partial Readout command does not enable the partial readout; you still have
to click the Partial Readout On/Off button afterwards.
Snapshot
Use the Image>Snapshot command acquire a single images and to make it available for the image-anal-
ysis software.
You may also You can also click the Standard (button bar) > Snapshot button.
Available The command is not available if the active image buffer is write protected.
If the live mode was enabled with the Image > Acquire command, it will
automatically disabled with the Snapshot command.
Before acquiring an im- You should first check the live mode if you are not certain if the object to be
age acquired is positioned, focussed, or illuminated correctly. The illumination of
the object, the setting of the diaphragm, as well as the setting of brightness
and contrast are important for the quality of your images.
19
Use the intelligent Exposure procedure to make acquisitions with little effort.
intelligent Exposure largely makes the acquisition procedure automatic. The

exposure times for the live acquisition and the snapshot are automatically
set independent of one another so that the dynamic range of the camera are
fully taken advantage of. The optimization of the exposure time occurs con-
tinually and automatically.
Histogram-Analysis intelligent Exposure analyzes the real time histogram during a live acquisi-
tion in order to optimize the exposure times.The automatic gain display and
the exposure time are set so that the signals use one of the dynamic ranges
set by the user.
Calibration intelligent Exposure arranges linear correlation between the exposure time
and the mean gray value for calculating the optimal exposure time. In order
to take advantage of this correlation, the camera's dark current must be
known as an offset of the linear function. Thus, the first step to working with
Related Topics
Use automatic gain display 15
Dynamic limit in % 22
ColorView II • intelligent Exposure
intelligent Exposure is measuring this dark current via the acquisition of an

image using a darkened camera. When first starting intelligent Exposure,
you will be automatically asked to carry out this calibration.
This calibration step measures the camera's dark current, in order to make
an offset correction possible. After successful measurement of the dark cur-
rent, intelligent Exposure asks you to do the next step.
The second calibration step consists of measuring the signals in various res-
olution settings. This detects the camera's sensitivity. A changing live-image
is displayed in this step.
You must execute the calibration anew should you change cameras, i.e. by
switching to another input channel. You have access to this function by us-
ing the Calibration... button located in the intelligent Exposure > Options di-
alog box which is described further down.
intelligent Ex- Click the intelligent Exposure button to be able to make acquisitions using
posure this method. The intelligent Exposure dialog box is opened.
20
Exposure times The exposure times ascertained by intelligent Exposure are displayed in the
dialog box's status bar. The term Live Exp. (Live Exposure Time) stands for
the exposure time of the live acquisition, while Snap Exp. (Snapshot Expo-
sure Time) stands for the exposure time of the snapshot. Independent of the
calculated exposure time for the snapshot, the exposure time for the live ac-
quisition cannot exceed 125 ms. In doing so, a quick live-image is guaran-
teed which simplifies the microscope's settings when the camera is running.
It is possible that the live-image is very dark, should the changes you made
to the setting result in very long exposure times for snapshots. Please alter
the microscope settings accordingly in such cases.
Acquisition Click the Acquire button to start the live acquisition using intelligent Expo-
sure. The live-image is displayed in the image document. You can make the
necessary settings (focus, stage settings) with the help of this live-image.
Characteristics of the The speed of the live-image has absolute priority when using intelligent Ex-
live-image posure. For this reason, use the camera's automatic mode-switch to in-
crease the live-image's frame rate. These changes, however, do not affect
the snapshots which were made with intelligent Exposure.
Snapshot Click the Snapshot button to initiate the acquisition of a snapshot. intelligent
Exposure acquires an image and saves it to the active image buffer. A hint
box informing you about the acquisition process appears if the exposure
time of a snapshot exceeds one second.
Note Use the Special > Preferences... command to be able to acquire numerous
snapshots simultaneously without overwriting them. Enable the Images tab.
Select the Image buffers (All) entry from the. Image acquisition> sequence
list. A new image buffer is automatically enabled after the acquisition of a
snapshot.
Exposure time correc- The Exposure time correction slide control enables you to manually influ-
tion ence the exposure time for snapshots calculated by intelligent Exposure.
Position ’0’ on the slide control does not influence the exposure time which
was automatically calculated by intelligent Exposure. Move the Exposure
time correction control to the right in order to increase the exposure time, or
to the left to shorten it. The exposure times for live acquisitions and snap-
shots are equally altered during correction, as long as the exposure time for
snapshots is lower than 125 ms. The exposure time for the live acquisition
remains unchanged for exposure times exceeding 125 ms.
Note Moving the control to the left only leads to a hardly visible change in bright-
ness in the live-image. This is due to the automatic gain display function
which is always enabled when using intelligent Exposure.
21
Resetting the Exposure Double click on ’0’ below the Exposure time correction slide control in order
time correction to use the automatically calculated exposure time values once again.
Exposure Time Click the Exposure Lock button to lock the current exposure time for intelli-
Lock gent Exposure. As of now, no more alterations will be made to the exposure
time. Click the button anew to return to the dynamic adjustment of the expo-
sure time.
Preview Click the Preview button to use the setting for snapshots with live-images as
well. Now the resolution and exposure time for snapshots will be used. Click
the Preview button anew to once again use the settings for the live acquisi-
tion.
White Balance Click the White Balance button to carry out a special white balance when us-
ing intelligent Exposure. In the image document, move the ROI's red rectan-
gle to a white or uniformly gray position on the specimen. Change the size
of the ROI by keeping the right mouse key depressed and moving the
mouse. Rightclick to confirm position and size of the ROI.
Options Click the Options button to set the options for intelligent Exposure. The in-
telligent Exposure dialog box is opened.
ColorView II • intelligent Exposure
Dynamic limit in % Set the dynamic range in which intelligent Exposure works with the Dynamic
limit in % slide control. Move the controller to the right to increase the dy-
namic limit. This setting results in lighter images with a higher contrast in
darker areas of the image. Move the controller to the left to decrease the dy-
namic limit. This setting results in darker images with a possible better con-
trast in light areas of the image. This effect is not distinctly evident when us-
ing color mode.
Note Due to the calculation procedure, setting the Dynamic limit in % to 90 can
lead to an overflow. You should reduce the Dynamic limit in % for acquisi-
tions whose high gray values contain important information for the image, for
example in fluorescent microscopy.
Max. exposure time in Set the highest exposure time to be used for snapshots using the Max. ex-
s posure in s slide control. You can also enter the desired text directly into the
text field. This value determines the maximum exposure time for snapshots
which intelligent Exposure can use. The maximum value which can be set
here depends on the camera being utilized. A warning occurs and the pic-
tures become darker should intelligent Exposure calculate a higher expo-
sure time than determined by Max. Exposure in s.
Max. exposure preview Set the highest exposure time to be used for live image acquisitions using
in s the Max. exposure preview in s slide control. You can also enter the desired
text directly into the text field. This value defines the maximum exposure
time for live image acquisitions which intelligent Exposure can use. The
22
maximum value which can be entered here, depends on the camera being
utilized. A warning will appear should this value be exceeded by the current
illumination conditions.
Resolution Select the size of the images to be acquired in the Resolution group. In doing
so, the resolution of the images changes, however not the image segment
shown.
Acquisition Select the image size for the live acquisition from the Acquire list. Next to the
full resolutions, you will also find lower exposures with half and a third of the
pixel count in each direction available. intelligent Exposure uses this expo-
sure.
Snapshot Select the image size for snapshots from the Snapshot list. Next to the full
resolutions, you will also find lower exposures with half and a third of the pix-
el count in each direction available. intelligent Exposure first sets the full ex-
posure automatically.
Keep live after snap- Select the Keep live after snapshot check box, in order to continue the live
shot acquisition directly after a snapshot. A snapshot is created in the live acqui-
sition's image buffer and a new live acquisition is started in the next image
NetCam
buffer defined in the sequence. Keep live after snapshot is only available if
you have selected an entry other than None in the Sequence list located in
the Special > Preferences > Image > Image acquisition dialog box.
Calibration... Click the Calibration... button to begin the camera's calibration process. The
procedures for calibration are the same as the procedures located in the
section entitled "Calibration".
NetCam
You can transfer the ColorViewII live-image via the intranet or internet to
other computers using the add-In NetCam.
This enables you make the images available for your colleagues directly
during acquisition. Thus, interactive work over large distances is made pos-
sible using simultaneous communication, e.g., telephone. The images are
compressed so that they can be transferred quicker.
Transferred images The live-image is directly transferred. Based on the speed of your network
connections and the attained refresh rate during acquisition, you will get the
impression of a live-image. In an internal company network, a direct backup
coupling from observer to user is possible without any elapsed time.
NetCam can also transfer a fixed image, whereby in this case the image's
overlay is also transferred.
Definition The Server is the computer on which both the image-analysis software and
the NetCam are installed. The image acquisition is controlled from the serv-
er.
The Client is each computer which can access the server's images via in-
ternet. The client cannot control image acquisition.
Available A web server must be installed on your computer to make images available
via the internet. An appropriate program is automatically installed by Setup,
if you have selected the add-In NetCam. This step will be skipped should a
web server already exist on your computer.
Warning When using a computer connected to a network it is possible that the secu-
rity settings will not allow the use as a web server. Therefore, contact your
23
system administrator.
In the Module\NetCam subdirectory of your image analysis program, you will
find the file "NetCam_Info.pdf". This file contains technical explanations
about NetCam which you can give to your system administrator.
Installing the web server

The web server required for the use of NetCam is automatically installed during the setup.
The installation of the web server is started directly by setup. After having
agreed to the license terms, you can make further settings in a dialog box.
Related Topics
Calibration 19
ColorView II • NetCam in the intranet
You can simply adopt the default settings.

Complete, Custom We recommend the complete installation of the web server.
NetCam in the intranet

The transfer of images within the company network or intranet generally
does not require any further actions.
Enabling access During installation, setup will create a link with the name NetCam.html in the
\Module\NetCam subdirectory below the working directory of your image-
analysis software. Send this link via email to colleagues you would like to
give access to your images. The easiest way to do this is with the following
command: Send to > E-mail receiver in the context menu of the link in Win-
dows Explorer.
Java Java Runtime Environment, Version 1.4.1_03 or higher is required to show
24
the images on the clients. When this requirement has not been fulfilled by
the client, the transfer of images cannot be carried out.
NetCam in the internet

When using NetCam, a direct communication takes place between the serv-
er and the client. If the communication goes beyond the company network,
you must, if need be, make changes to the security settings.
Adapting the security The adaptation of a network's security settings takes place via the respec-
settings tive system administrator. The necessary technical and security require-
ments for the use of NetCam are outlined in a separate file entitled
"NetCam_Info.pdf" which was saved by setup to your system's image-anal-
ysis software \Module\NetCam subdirectory. Please give this file to your
system administrator so that he can make the appropriate adaptations.
IP Address The server's internet address must be entered on the client's page. This ad-
dress is available in the well-known alphanumerical form ("MyComput-
er.OurCompany.de") and in the numerical form ("62.180.61.137"). The nu-
merical form of the address is more reliable, especially if your system admin-
istrator has to alter the network settings.
Using NetCam
Determining the IP ad- You can find out your computer's IP address from your system administrator
dress or determine it yourself.
How to... This is how you determine your computer's IP address:
1) Select the operating system command Start > (All) Programs > Acces-
sories > Command Prompt.
" The Eingabeaufforderung console box is opened.
2) Enter the ipconfig command with your keyboard and use the enter key.
" Data about your computer's network connection are shown in the
console box.
3) Note the entry in the IP address row (192.168.118.175 in example

above).
4) Make this information available to the users on the side of the clients.
Java Java Runtime Environment, Version 1.4.1_03 or higher is required to show
the images on the clients. When this requirement has not been fulfilled by
the client, the transfer of images cannot be carried out.
Using NetCam
The use of your image-analysis software remains the same when using Net-
Cam. You can use all of the functions as always. To transfer the images to
25
other users and to be able to control the transfer, use the buttons located in
the NetCam button bar.
NetCam on/off Click the NetCam On/Off button to start or end the transfer of images with
NetCam.
The transfer of live-images begins as soon as NetCam is activated. The
transfer continues until you deactivate NetCam or close your image-analysis
software.
NetCam Set- Click the NetCam Settings button to set image options and to administer the
tings clients which are to have access to your images. The NetCam Settings dia-
log box is opened.
ColorView II • Using NetCam
Connections Use the Connections tab to administer the clients which are to access your
images.
Connected Clients In the Connected Clients list you will see all of the clients which are currently
connected with your computer. You can mark one of these clients to discon-
nect the connection to it or to refuse a connection in the future.
Disconnect Click the Disconnect button to disconnect the connection to the marked cli-
ent. The client can once again make a connection.
>> Click the >> button to include the marked client in the Banned Clients lists.
The corresponding client is automatically disconnected.
Banned Clients In the Banned Clients list you will see all of the clients who you have forbid-
den access to your image-analysis software. From these computers it is im-
possible to load and view the images made available by NetCam.
Add new... Click the Add new... button to add computers or entire groups of computers
to the Banned Clients list. The Enter IP(s) or a domain dialog box is opened.
26
Domain name Enter the alphanumerical name of a domain into the Domain name field if
you want to block the access of computers within this domain. For example,
enter the domain name foobar.com to hinder the transfer of the images to all
the computers within the domain foobar.com.
Procedure NetCam rends a query to the Nameserver to get the numerical IP address
belonging to the domain provided. You will receive a message, should the
query lead to an error. The provided domain will not be placed on the
Banned Clients list.
IP Address(es) You can provide a numerical IP address or an entire address range in the IP
adresse(s) group to exclude the computer in question.
From, to Enter the limit of the address range to be excluded in the from and to fields.
Usage in Black & White mode
Image options In the Image Options tab you can set the quality of the transferred images
and thus influence the transfer rate.
Image quality Use the Image Quality slide control to control the compression of the imag-
es. Move the slide control to the right to maintain a high image quality. The
data amount of the transferred images increases, whereby the transfer rate
decreases. Move the slide control to the left to increase the compression.
The image quality decreases, whereby the transfer rate increases.
Image quality and data A single image of ColorView II can have the size of up to 12 MByte. Images
transfer rate of this size take a long time to transfer to a client even with a very fast net-
work connection; the live-image effect is thus lost. Therefore the images are
compressed for transfer. The used algorithm is equivalent to that used with
JPEG images. Thus, high compression rates are achieved with decreased
image quality.
The amount of compression achieved is based on the image type. Images
with large, unison colored areas can be compressed more than detailed im-
ages without these characteristics.
Usage in Black & White mode

Some changes result when using a black & white camera or a color camera
27
in black & white mode. This chapter describes the control elements in the
Camera Control dialog box which are different from those in color mode.
Available A separate input channel is required in black & white mode. Setting up this
input channel will be described in the section entitled "New Channel".
Enabling Black & White Use the Image > Set Input... command to select the input channel which was
mode created for the black & white mode.
Related Topics
New Channel 30
ColorView II • Usage in Black & White mode
Gamma Correction
In areas with rather high and rather low intensities, cameras can come to
non-linearities which can decrease the contrast of these areas. You can use
a gamma correction to make a physiological correction.
Gamma Setting Click the Gamma preferences button to set up a quick gamma correction
during a live acquisition. The Gamma Setting dialog box is opened.
Move the Gamma slide control to make a gamma correction of the live-im-
age. Move the controller to the left (Gamma < 1) to make the entire image
darker; whereby the brightness in light areas will be increased. Move the
controller to the right (Gamma 1) to make the entire image lighter and to in-
crease the contrast in the dark areas of the image.
Note The image will be displayed using an LUT (Look-Up Table), or in other words
is converted into a false-color image. For these reasons, not all the analyses
are possible on the image any longer.
Ref. The Ref. LED (light-emitting diode) has no meaning for ColorView II.
Display and usage of the acquisition with Real time FFT

Available Real time FFT is only available for the camera in black & white mode. To be
able to utilize Real time FFT during a live acquisition, you must select the
Image > Configure input > Display > Live Overlay check box.
Enabling Real time FFT Real time FFT can be activated in the input channel. To do this, select the
Image > Configure input > Display > Real time FFT dialog box.
Real-time Fast Fourier Transformation (RTFFT) calculates the acquired im-
age's Fourier image in real time (the absolute value of the Fourier trans-
28
forms). The Fourier image is continually updated during a live acquisition. It

can serve as a quantitative display of the image quality. For example, if you
blur the microscope image, the high frequencies disappear in the Fourier im-
age (i.e. the structures which are located farther away from the center).
Camera Set Up
The Fourier Image of a Cat-

alase 2-D Crystal.
The points represent period-
ic structures in the micro-
scope image. The closer
such a point is to the center,
the smaller the spatial fre-
quency of the correspond-
ing periodic structure, i.e.
the larger the periodic dis-
tances in the microscope
image. (The cross arises
due to fringe effects.)
The RTFFT Control dialog box is available in order to optimize the onscreen
presentation of the Fourier image:
Contrast The Contrast slide control enables you to alter the contrast of the display in
the RTFFT Display window. Move the controller to the right to increase the
contrast and to the left to decrease it. Should you have selected a Display
palette the colors will change in the RTFFT Display window. You define the
29
palette in the input located on the Display tab.
Brightness The Brightness slide control enables you to alter the brightness of the Real
time FFT display in the RTFFT Display window. Move the controller to the
right to increase brightness. Move the controller to the left to decrease
brightness. Should you have selected a Display palette the colors will
change in the RTFFT Display window.
Camera Set Up
Status after installation An input channel is automatically created when installing the software, im-
mediately enabling you to make image acquisitions. This channel is, in many
cases, adequate for all demands.
Apart from the actual installation, the configuration of one or more input
channels is also part of the installation required for setting up the camera.
ColorView II • Software Camera Control
Software Camera Control

After successful installation, you can configure the camera for using the image-analysis software.
Logical input channels The ColorView software complies with the logical input channel concept.
This means the camera is as easy to operate as a regular video camera if it
is hooked up to the microscope and computer, and if the software has been
installed and configured. No additional menu appears in the user interface's
menu bar. When you have selected the Set input... command located in the
Image menu, you will realize that you have additional logical input channels
at your disposal which can be set up to fit your needs.
Configuration via tabs Channels are configured via the inputting of suitable values in various tabs.
The 3 tabs relevant for ColorView II will be dealt with in-depth.
Logical input channels contain, for example, the information about how large
an image format is, and what is to happen with the image signal before it
reaches the image buffer. Each channel must be calibrated separately. A
channel for image acquisition via ColorView II is usually called "ColorView II
FW...".
Setting up an input chan- If you have to set up a new input channel, you can create a copy of an al-
nel for black & white use ready existing input channel and adapt it, or create an entirely new input
channel. A new input channel is required for use in black & white mode.
Set Input Use the Image > Set Input... command to select a new input channel or to
create a new one. You can also depress the [F6] key. The Set Input dialog
box is opened.
The Set Input dialog box
with an already defined in-
put channel.
30
New Channel Click the New Channel button to create an entirely new input channel. The
Select device dialog box is opened. All connected image creating devices
are listed in the Available devices list.
Select Device dialog box

with a connected ColorView
II camera.
The abbreviation BW in the
last line marks this camera's
black & white mode.
Select the desired camera mode (color or black & white) in the Available de-
vices list and click OK in order to create a new channel.
Creating additional input Should you want to create additional input channels, for example to have dif-
channels ferent configurations immediately available for repeating tasks, use an al-
ready existing input channel.
Duplicate Click the Duplicate Channel button to make a copy of the active input chan-
Channels nel. The new input channel gets the same name as the channel from which
it was copied and receives a sequential number. The newly created channel
is automatically enabled.
Configure In- Click the Configure Input button to adapt the settings for the new channel.
put The Configure Input dialog box is opened. You can also open the dialog box
using the Image > Configure Input command or by double clicking the cam-
era name located in the status bar.
The Input tab
31
Exposure Enter the number of microseconds the CCD chip of the camera is to be ex-
posed into the Exposure field. You can also use the arrow key (to the right
of the field) to select this value. The chapter entitled "Camera Control" ex-
plains how you can change the value in this field with the help of the Camera
Control... command interactively while viewing a live-image.
The smallest possible exposure time is 100 µs and the largest is 160 s. The
dynamic breadth from ultra-short to long-period exposure enables the acqui-
sition of high-intensity, brightly exposed images, as well as the fluorescent
microscopic reproduction of very dim glowing structures.
Resolution You can choose three different resolutions possible with ColorView II from
the Resolution list. The adjustable resolution in ColorView II substitutes bin-
ning.
Online shading You can set the camera function with the same name in the Online Shading-
correction correction group.
Activate Select the Activate check box to use the online shading correction. The on-
line shading correction will immediately be used should correction images
already exist for the current settings. Should there not be appropriate cor-
rection images available, the application will automatically start the wizard
for the acquisition of correction images.
Illumination method Select the procedure used by you from the Illumination method list.
Sharpen filter Select the Activate check box of the Sharpen filter group to enhance the im-
Activate age sharpness. This sharpen filtering takes place in real time during image
acquisition. When applying the sharpen filter, the numerical value in the
Sharpen field determines whether or not the image’s contours appear sharp-
er or softer.
Raising sharpness will accentuate edges, but also brings out image noise
more.
Please note that when you use the sharpen filter you not only alter the mon-
itor display, but the actual image information as well. This means that when
you are planning on quantitatively evaluating your images later, you should
not use the sharpen filter. The Oper menu provides you with commands you
can use at any later time for adjusting an image’s sharpness.
Sharpen Enter the desired sharpness parameter into the Sharpen field. You can also
32
use the arrow key (to the right of the field) to select this value. Values rang-
ing from -30 to 30 are possible, whereby the default value is 10. The image
appears smoother when using negative values and sharper when using pos-
itive values. Entering a value of 0 will result in the sharpen filter having no
effect at all.
Convenient alternatives for activating/deactivating the sharpen filter, as well
as for adjusting sharpen filter parameters, are the Sharpen Filter On/Off but-
ton and the Sharpen Filter Settings button - both in the Camera Control dia-
log box.
Mirror The Mirror group enables you to determine if the camera's image is to be
mirrored directly into the program during transfer. Both mirror functions can
be combined with each other.
Horizontal Select the Horizontal check box to mirror the image horizontally during ac-
quisition.
Related Topics
Camera Control 12
Acquiring Correction Images 7
Vertical Select the Vertical check box to mirror the image vertically during acquisi-
tion.
Info Click the Info... button to have some device data, as well as the current tem-
perature of the CCD chip and the camera housing shown in the ColorView
II FW Information window:
Device Name The Device Name relates to the inner construction of the camera and is
specified by the manufacturer.
Devices ID The Device ID is the physical identification of the camera for the operating
system. It is in accordance with the device's serial number.
Firmware Version The Firmware is the cameras software for your operation.
Serial Number The serial number serves as a clear identification for each camera pro-
duced.
Camera Type The Camera Type is the exact identification for the camera model.
DLL Version The DLL Version is the version number of the camera's driver.
SYS Version The SYS Version is the version number of the camera's device driver. DLL
and SYS Version are identical.
33
Temperature The cameras are equipped with temperature sensors. The temperatures are
continually measured.
CCD Chip CCD Chip is the CCD chip's temperature. An increase in noise is the result
of high temperatures of the CCD Chip. The default value for trouble-free op-
eration is 10°C.
Housing Housing is the temperature of the cameras housing. The cameras housing
should not exceed 40°C.
The Display tab
Over exposure The Over exposure group enables you to determine if and when you should
be warned of an overexposure. The software continually checks the image
intensity in live mode, by analyzing the image's gray-value histogram in real
time. You will be warned if the live-image is overexposed. The warning will
also appear following acquisition of an overexposed single image.
Display warning Select the Display warning check box to have a warning message shown if
the image is being overexposed:
34
In this case, reduce the exposure time till the warning message disappears.
Overflow An image is overexposed when a certain percentage of its pixels attain the
maximum gray value possible. This percentage is entered into the Overflow
field.
Automatic gain display Use the functions in the Automatic gain display group to have the image al-
ways displayed optimally onscreen, no matter what the actual lighting con-
ditions are. The system assesses the current gray-value histogram - in real
time - in order to obtain optimal display of the live-image on the monitor.
Note that the automatic gain display not only effects the monitor display, but
also effects the image information. If it is activated, the bit-depth of the ac-
quired images is reduced from 3 x 12 Bit to 3 x 8 Bit with the help of the op-
timized lookup table. If automatic gain display is not activated, the 4 lower
bits (from the 12 bits of each component color) will simply be clipped.
Activate Select the Activate check box to enable the automatic gain display. The ef-
fects of this check box is the same as clicking the Use automatic gain display
button located in the Camera Control dialog box (<S. 15). The Activate
check box is not available if the check box with the same name in the Fixed
Scaling group is enabled.
Left overflow When the automatic gain display is active, image intensity for the image on
Right overflow the monitor will be spread between the gray values Gmin and Gmax. You de-
termine the values of these Gmin and Gmax gray values in the Left and Right
overflow fields: this is where you enter the percentage of pixels with the low-
est gray values which are to be displayed black, and which percentage of
the highest gray values are to be displayed white.
This is how you clip gray values that are to be ignored at both the upper and
lower ends of the gray-value histogram. Make use of the overflow values so
that individual pixels or minute image areas that are too bright or too dark do
not influence the automatic gain display.
Fixed scaling Instead of using the Automatic gain display group, you can use the Fixed
scaling group. The group whose Activate check box has been selected will
be the one available - the other will not be available.
The functions of the Fixed scaling group are for manually influencing on-
screen image display. You select definite limits - Gmin and Gmax - for the
gray-value range that you wish to have optimally spread for onscreen dis-
play. Any gray values occurring outside this defined gray-value range will
appear either black or white.
Application This is how you can, for example, improve the visibility of the specimen
structures of interest to you - by having dark areas of the specimen dis-
played entirely in black.
Please note that this manual presetting of the gray value, like the automatic
gain display, not only effects the monitor display but also the image informa-
tion.
Activate Select the Activate check box to switch on the manual definition of the gray-
value range to be displayed.
Left limit The left limit can only be set in black & white mode; using it with color cam-
eras would lead to a color tinge. Enter the value of the left limit for the fixed
35
scaling into the Left limit box.
Right limit Enter the value of the right limit for the fixed scaling into the Right limit box.
Its function is equivalent to the Right limit parameter, which you can influ-
ence by clicking the Manually adjust fixed scaling button located in the cam-
era controls dialog box.
Online histogram Select the Online histogram check box to be able to keep an eye on the
gray-value distribution during image acquisition. During image acquisition, a
window showing the current histogram will appear automatically. This histo-
gram will be continually updated.
Related Topics
Manually adjust fixed scaling 16
Online histogram while op-

erating in color mode
(above) and in black & white
mode (below).
Minimum, mean, and maximum gray values of the image will be shown be-
neath the histogram. Percentages are in relation to the maximum obtainable
gray value. The maximum obtainable gray value when using ColorView II
(with its 12-bit depth) is 4095. This gray value is equivalent to 100%. A mean
value of 33,6% when operating in black & white is equivalent to a gray value
of 1376.
During a live acquisition, you can turn on or off the Automatic Gain Display
with the buttons in the online histogram, as well as activate and set the fixed
scaling.
Setting the fixed scaling You use the mouse to set the limits of the fixed scaling in the online histo-
via the online histogram gram. Move the pointer over the blue (lower limit) or red (upper limit) vertical
line in the histogram. The pointer turns into a horizontal double arrowhead.
Move the mouse with the mouse button depressed to the left or right to either
increase or decrease the limit values. The changes can be viewed directly
36
in the live-image.
Real time FFT The Real time FFT check box is only available in black & white mode and
with black & white cameras. In order to be able to use the Real time FFT dur-
ing live acquisition, you must select the Live Overlay check box. An expla-
nation of the effects of the enabled Real time FFT can be found in the sec-
tion entitled Display and usage of the acquisition with Real time FFT.
Live overlay Select the Live overlay check box if you want to use overlays in live-images.
Live overlays are absolutely necessary for partial readout, white balance, or
Real time FFT.
An image’s overlay contains information in the form of text, markers, or other
graphical elements. The overlay can be displayed together with the corre-
sponding image without the actual image information being affected. The
live overlay must be enabled in order to make Real time FFT or partial read-
out possible.
Related Topics
Use automatic gain display 15
Use fixed scaling 16
Display and usage of the acquisition with Real time FFT 28
Partial Readout 18
The Live overlay check box is only available if your display board features a
so-called hardware overlay which is supported by Microsoft DirectX. Almost
any up-to-date AGP display board provides you with that feature. You can
look at the System Info dialog box to check whether your display board of-
fers this feature. To do this, select the About... command located in the ?
menu and click on the System Info... button. The System Info dialog box will
be opened. Click on the "DirectX" entry in the list on the left-hand side to
have the desired information appear in the list on the right-hand side.
Image scaling Select one of the four selections for displaying an acquired image in a View-
port in the Image scaling list.
Underscan Select the Underscan entry to have the whole image displayed. The system
will select the largest zoom level - 25%, 50% or 100% - where the whole im-
age can still be displayed within the Viewport. In certain cases not all of the
Viewport’s available area will be used.
Overscan Select the Overscan entry to ensure that all the Viewport’s available area will
be used. The system will select the smallest zoom level - 25%, 50% or 100%
- where the image fills the whole Viewport. In certain cases parts of the im-
age will not be visible.
Adjust to viewport Select the Adjust to viewport entry to have the image size adjusted to opti-
mally fit the Viewport. The image will be adjusted to fit the current size of the
Viewport.
Full size (100%) Select the Full size (100%) entry to have the image displayed without any
zooming. Only the left upper corner of the image will be shown if the View-
port is smaller than the image.
Histogram calculation Select the pixels to be taken into consideration for calculation of the gray-
value histogram in the Histogram calculation list. There are three options:
Full image If you select the Full image entry, the gray values of all pixels will be applied
to histogram calculation. This is the most commonly used setting when us-
ing light microscopy.
Crosshairs If you select the Crosshair entry, only pixels of a particular image area are
used in the calculation. This image area is shaped like cross hairs that are
centrally positioned on the image. The vertical and horizontal segments of
these cross hairs are respectively just a few pixels wide and cover just about
37
the entire height and width of this image. Crosshair is the default setting
when using transmission electron microscopy.
ROI Select the ROI entry to carry out the optimization of contrast and exposure
time based on a rectangular frame.
These settings effect all real time functions which use the gray-value histo-
gram: over exposure warning, Automatic gain display, and the Online-Histo-
gram.
Application We recommend selecting Crosshair if you wish to speed up histogram cal-
culation. You can speed up the frame rate of the live-image depending on
what real time functions you have activated. You should however make sure
that the pixel gray values around the middle image line and around the mid-
dle image column are representative for the gray values of the entire image.
If this is the case, you will not notice any difference compared to the Full im-
age option for the over exposure warning, Automatic gain display, and the
online histogram.
Display palette You can select a false color lookup table from the Display palette list to color
the acquired image. The Display palette list is only available in black & white
mode.
- none - Select the entry - none -, not to use any palette for the image display.
Adjust, Dither, Gamma, The entries in this list are the names of various lookup tables which can be
Mixmap Rainbow, So- used for the image display on the monitor.
laris, Thermal
The Format tab
Active area The display in the Active area field shows how large the acquired image is
using the present settings with regards to the largest possible image size. If
you make changes to the clipping, you can directly view their effect here on
the active area.
Image format You can choose a means of image acquisition from the Image format list.
38
The entries in this list correspond to predetermined settings for the clipping.
In addition to the predetermined list elements, you can also select from the
settings which you defined.
Binning The Binning function is not enabled when using ColorView II. Use the Res-
olution function located on the Input tab instead.
Clipping border (pixel) When clipping is used, the edges of the images are clipped. Hence, clipping
does not have any effects on the resolution and brightness as well as the
required exposure time.
Left, Right, Top, Bot- The fields entitled Left, Right, Top and Bottom show the amount of pixels
tom that are to be clipped from each image edge. Each individual value can
reach from 0 (no clipping) to almost the entire image in each respective di-
rection (complete clipping). If a value for the clipping already has been set,
the setting possibilities for clipping the opposite edge reduce themselves re-
spectively.
Related Topics
Resolution 32
Interactive Click the Interactive button to be able to place the clipping interactively into
the image document. Move the pointer with the mouse button depressed to
change the size of the area to be acquired. The actual acquired area is
marked by a red frame. Move the mouse to alter the position of the marked
area within the image document. Rightclick once to fix the area to be ac-
quired.
No Clipping Click the No Clipping button to return the values for Left, Right, Top and Bot-
tom back to 0, thus disabling Clipping.
39
ColorView II • Appendix I: Microscope Settings
Appendix I: Microscope Settings

Your digital camera can only achieve high image quality if the microscope
has been optimally set. A subsequent correction to the software can never
correct deficiencies in the image resulting from faulty settings made to the
camera and microscope.
The optical system microscope offers numerous setting possibilities. In or-
der to attain best acquisition results, the settings must be made precisely.
In spite of varying models and observation possibilities, the microscope's
image producing components are in accordance with each other.
A microscope basically consists of the optical components: projection objec-
tive and magnifier. The projection objective is the objective with which the
observation is to be carried out. The eyepiece takes on the function of the
magnifying glass. An additionally important component is the illuminator.
This illustration clarifies the
basic assembly of the com-
ponents of a microscope in
transmitted-light mode.
40
Illumination
Light is a decisive medium for creating images with a microscope. As a result, the light, i.e. illumination,
plays an important role in creating the image in the microscope.
The goal of the illumination settings is the equal distribution of light on the
sample with parallel light rays falling on the object. The illumination must be
bright enough to make all of the image details visible; however over-illumi-
nation leads to images being too bright and low-contrast. In addition, the col-
or of the light has a strong impact on the sample's color rendering.
Lamp Selection The tungsten-halogen lamps and gas discharge lamps are the most com-
monly used lamps in microscopy. The light creating processes of both lamp
types are fundamentally different.
Illumination
Tungsten-halogen Tungtsten-halogen lamps comply with their functionality to a conventional

lamps light-bulb. Electricity flowing through a metal wire heats the wire to the extent
that light is produced (thermal emitter). In doing so, the wire emits a contin-
uous spectrum, hence a broad spectral band is emitted. One can control the
brightness as well as the color (color temperature) of the lamp by controlling
the electrical current. Tungsten-halogen lamps can be widely used in mi-
croscopy, especially when acquiring color images.
Gas discharge lamp Gas discharge lamps stimulate gas at the atomic level using strong electrical
fields. The charged atoms return to their initial state and, in doing so, give
off the energy which initially charged them in the form of electromagnetic ra-
diation: light. The resulting photons each have characteristic energy which
is typical for the gas being used. As a result, this type of lamp does not emit
a broadband spectrum like the halogen lamp, but instead emits discrete
wavelengths. This fundamental physical process cannot be externally influ-
enced; hence, such a lamp's perceived color is independent of the operation
conditions. Because of this discrete spectrum, gas discharge lamps do not
emit a color-neutral illumination. Its adjustable high intensity light and the
emission of short-wave spectral bands required for fluorescent microscopy
is an advantage.
ND-filter Many microscopes can insert ND-filters into the light path, thus reducing the
intensity of the lamp's light. These ND-filters ensure a uniform reduction of
light intensity throughout the entire frequency range without altering the col-
or temperature.
Köhler Illumination
This illumination process which was described by Dr. August Köhler as early
as 1893, is still an indispensable prerequisite for successful analysis using
light microscopy. The Köhler illumination enables an optimal, homogenous
illumination of solely the part of the sample to be analyzed.
Condenser The condenser carries the light onto the sample. The main components are
the aperture stop with which the size of the illuminated area is set and the
condenser lens which is responsible for the parallel emitted light rays. The
condenser can be opened and closed. The condenser is height-adjustable
and horizontally adjustable.
41
Note Set the aperture stop to 2/3 of the entire opening and do not use it to reduce
the brightness. To do that, you should always use ND-filters or a lower lamp
voltage.
Principle of Köhler Illumi- You will find comprehensive descriptions about the principle of Köhler illumi-
nation nation in microscopy text books. We will only give you a brief overview about
some important settings you should know.
Focussing the Field Di- One vital step when setting up the Köhler illumination is the sharp view of
aphragm the field diaphragm at the specimen level.
1) Focus the object to be viewed.
2) Close the field diaphragm so that only a small bright spot is visible in
the eyepiece.
3) Focus the image in the field diaphragm by using the condenser's height
adjustment.
ColorView II • Objective, Total Magnification and Useful Magnification
Adjusting the Con- Now the illuminated area is optimally set.

denser 1) Open the field diaphragm so that its edge is still completely within the
visible area.
2) Center the image of the field diaphragm using the condenser's adjust-
ment screws.
" A precisely adjusted condenser guarantees that only the area of the
specimen to be viewed is illuminated.
3) Repeat steps 1) and 2), until the edge of the field diaphragm is no long-
er visible in the visual field.
Adjusting the Lamp You can generally skip adjusting the lamp with modern microscopes, provid-
ed that the lamp is located in a frame and the bulb is in a centered socket.
Objective, Total Magnification and Useful Magnification

There are no general rules for which objective you select; one should always
keep the specimen and reason for observation in mind.
Terminology The two most important parameters of an objective (magnification and nu-
merical aperture) are engraved in the objective. First the magnification and
then the numerical aperture which is separated by a diagonal slash. Hence,
the code 40 / 0,65 marks an objective with a magnification of 40X and the
numerical aperture of 0,65.
Magnification An objective's engraved magnification is considerably important for the total
magnification. However, one should note that the range of useful magnifica-
tion should not be surpassed.
Numerical Aperture The numerical aperture is the characteristic of an objective which deter-
mines its capacity. The numerical aperture can also be found engraved into
the objective. It is based on the refraction index of the lenses being used as
well as the objective's angular aperture. The manufacturers are narrowly
limited when dealing with the refraction index, whereas the objective's angu-
lar aperture can be increased by using appropriate measures. To achieve
this, a fluid with a high refraction index is placed between the cover slip and
the objective. The immersion objectives are accordingly marked and should
only be used with the appropriate immersion fluid.
42
Total magnification The system's total magnification, to which an installed TV adaptor contrib-
utes, is based on the actual size of the final image. This can be the image
on the monitor, as well as the image printed on paper. To determine the total
magnification, it is best to use a stage micrometer which can be shown on
the monitor or the printout. You can determine the total magnification by di-
viding the length of the object shown by the set length on the micrometer.
Useful Magnification The term useful magnification specifies a total magnification of the optical
system by which full resolution is used without having a negative effect on
contrast or sharpness. The useful magnification of the optical systems mi-
croscope depends on the numerical aperture being used ( engraved on the
objective). The useful magnification is in the range of 500x to 1,000x the nu-
merical aperture.
Empty Magnification A loss of contrast and sharpness is the result if total magnification is larger
than the useful magnification. Check to see if replacing the TV adapter with
a 0,5 or 0,63 model sufficiently reduces the total magnification. If this is not
possible, you can also use an objective with the same magnification, but
with a higher numerical aperture.
Appendix II: Camera Installation
ColorView II
Appendix II: Camera Installation

Warning Please read the following carefully before attempting installation of
camera.
Requirements
The ColorView II can be used under the operating systems Windows 2000
and Windows XP. The installation procedure is described separately for
these two operating systems.
Installation sequence In order to avoid problems, you’ll have to follow the steps of the procedure
in the order given here.
Warning Install the image-analysis software first before connecting the camera
Windows 2000 to the FireWireTM board.
1) Check whether your computer system has a FireWire interface on-

board or a FireWire board installed.
2) Install the FireWire board provided with the camera.
3) Install the image-analysis software on your computer.
4) Install the camera and connect it to the computer system.
What if my computer sys- If your computer system already has a FireWire interface onboard or a sep-
tem already has a arate FireWire board installed, you do not need to install the board provided
FireWire interface? with the camera. If you are not sure whether a FireWire interface is available,
open the Windows Device Manager via Start > Settings > Control Panel >
System > Hardware > Device Manager....
43
An entry called ’IEEE 1394 Bus host controllers’ shows that the needed
FireWire interface is available.
ColorView II • Appendix II: Camera Installation
General Warnings!
• CCD image sensors are easily damaged by static discharge. Do not
disassemble the camera housing.
• Do not attempt to touch the internals of the camera with your bare
hands.
• Do not touch cover slip (over the CCD chip) with your hand or have any
object come in contact with glass surface. Should dust stick to the cov-
er slip, blow it off gently with an air blower. (For dust stuck due to static
electricity, ionized air is recommended.)
• It is best to use a LensPen to clean the cover slip. Do this only when
absolutely necessary.
• Do not expose to strong light (sunlight) for long periods.
• Exposure to high temperature or humidity will affect the characteristics
of the camera. Avoid storage or usage under such conditions.
• Do not subject the camera to too much mechanical shock.
• The FireWire cable plug is not symmetrical. You must make sure that
the cable plug is facing the same way as the camera socket when you
plug the cable into the camera jack.
• Do not disconnect the FireWire cable while the image-analysis soft-
ware is running.
The FireWire board IEEE-1394

Warning! • The components of these devices are easily damaged by static dis-
charge. Wear grounding wrist straps when handling them.
• Do not plug in or unplug the cards from the PCI slot while the PC is on.
• Please follow the installation manual in this booklet step by step to
avoid any incorrect connections. Incorrect connections may result in
serious damage to your PC system and/or the camera.
44
Installation Procedure - Windows 2000
Installation Procedure - Windows 2000

Installation of the FireWire board IEEE-1394
When is this necessary? This procedure is necessary only if the PC does not have a FireWireTM
interface on-board hardware configuration or if it is not yet equipped with a
FireWireTM board.
the FireWire board
How to install the 1) Turn off the PC and disconnect the power supply.
FireWire board 2) Remove the cover from the PC.
3) One PCI slot is required for the FireWire board. Insert the board into the
slot.
FireWire board installed
45
4) Replace the cover onto the PC.

5) You are now ready to first install the image-analysis software and then
the camera.
ColorView II • Installation Procedure - Windows 2000
Installation of the image-analysis software

Warning! The image-analysis software must be installed first before connecting the
camera to the FireWire board.
System requirements The Color View II camera requires the image-analysis software, (3.2 ver-
sion), and Microsoft Windows 2000 (or later). The image-analysis software
also supports multithreading on multiprocessors, thus making full use of the
power of the latest multi-CPU PC’s.
camera driver installation Installation of the SIS View FireWireTM camera driver is simply done via the
image-analysis software setup. You are only required to select FireWire and
specify the additional camera driver during setup.
Installation of the image- For installation, follow the instructions given on the monitor.
analysis software
In the image-analysis soft-
ware setup wizard select the
FireWire digitizer and select
your camera type. Select
the Firewire check box in the
Input Device (Digitizer)
Selection dialog box.
46
Then select the Soft Imag-

ing System manufacturer
entry and the Color View II
camera entry.
Precalibration You will be advised to carry out a precalibration of the camera when instaling
the image-analysis software.
In this dialog box you can
activate the precalibration.
47
Select the TV adaptor's magnification from the list which has the same
name. Should you not be working with a remote microscope, clear the Kam-
era soll mit Remote-Mikroskop betrieben werden check box. Then, mark the
checkbox for all mounted objectives in the Objektiv-Vergrößerungen des
Mikroskops list.
Note Should a magnification appear on the microscope objective which does not
apear in the list, a manual calibration must be carried out for this objective.
A guide is part of the image analysis system's step by step guide.
Note The image-analysis software can support several same or different types of
FireWire cameras simultaneously. You differentiate Color View II cameras
via their serial numbers when creating a new logical input channel via Image
> Set Input... > New Channel. A camera’s serial number is located directly
below the FireWire jack on the camera housing.
The camera’s serial number
is located below the
FireWire jack.
Installation of the camera and cables

Don’t forget... Install the image-analysis software first before connecting the camera to the
FireWireTM interface.
48
Installation of the camera and cables
How to install the camera 1) The diagram shows a typical installation of a light microscope. You
should have all required components ready at hand.
49
2) Remove the protective cap from the camera.
3) Firmly screw the TV adaptor (0,5x or bigger") onto the camera. This
should be done with the CCD-chip side of the camera facing down to
minimize the chance of dust landing on its surface.
4) Install this assembly onto the microscope's camera mount. Some mi-
croscopes may require additional optical elements installed between
the camera mount and the C-mount adapter. You may also need to
align the orientation of the microscope with that of the stage, and also
match the focus of the camera image with that of the eyepiece image.
These settings can be done later when you are able to obtain an image
on the screen.
5) Plug one end of the FireWire cable into the camera connector.
" The FireWire cable plug is not symmetrical. You must make sure
that the cable plug is facing the same way as the camera socket
when you plug the cable into the camera jack. The camera chip
may be ruined otherwise.
Make sure the cable plug is

facing the right way before
you plug it into the camera
socket
6) Plug the other end into one of the small connector sockets on the
FireWire board.
" There are two or more jacks depending on the FireWire board. You
can use any of them.
" The Digital Signature Not Found MS Windows dialog box(es)
appear on your screen. Confirm the message as often as neces-
sary until the dialog boxes disappear.
50
7) Open the Windows Device Manager via Start > Settings > Control
Panel > System > Hardware > Device Manager....
" The Device Manager dialog box contains the correct SIS FireWire
camera driver entry SoftHard Technology MV4.0 Camera. The
camera is ready for use.
Troubleshooting
The Windows Device Manager shows the IEEE 1394 FireWire board and the SIS View FireWire camera driver Soft-
Hard Technology MV4.0 Camera.
Troubleshooting
What do I do if the cam- An error message appears when starting the program, even though you
era cannot be found? have completely installed the camera and the image-analysis software:
51
The image-analysis software has not been successful in creating a connec-
tion to the camera. Confirm the message with OK.
Check to see if the connector cable between the camera and the computer
has been correctly plugged-in. Should the connecter be loose or not plugged
in correctly, correct this by firmly plugging them into the corresponding jack.
Restart the image-analysis software. Should the problem still exist, continue
with the following section.
What to do when the The error message No SIS-View FireWire camera has been found will also
camera has been con- appear when starting the program should the camera have been connected
nected before the image- before installing the image-analysis software. In this case, Windows 2000
analysis software instal- has already installed its own camera driver, which prevents the error-free
lation communication between the camera and the computer. Installing the image-
analysis software now does not install the correct camera driver since this is
forbidden by Windows 2000. You will notice that the camera is not detected
by the image-analysis software when starting the application.
If the camera is hooked up to the computer, you have to install the correct
camera driver from the software’s setup CD separately from the software
installation. Do the following:
1) Select the Start > Settings > Control Panel > System > Hardware >
Hardware Wizard... command.
2) Click on the Next button.

52
3) Select the Add/Troubleshoot a device option and click on the Next but-
ton.
Troubleshooting
4) Select the entry SoftHard Technology MV 4.0 from the Devices list and
click on the Next button.
53
5) Click on the Finish button.


54
7) Select the Search for a suitable driver for my device (recommended)

option and click on the Next button.
Troubleshooting
8) Insert the image-analysis software setup CD into your computer’s CD-

ROM drive. Minimize the appearing setup dialog box.
9) Make sure that only the CD-ROM drives check box is selected and click
on the Next button.
" Wait until Windows has found the correct camera driver. The
driver's data file is mv40dcam.inf. You will find it on the CD-ROM in
Directory\program\support\cameras\sis_firewire\drivers\win2k.
55

" The Digital Signature Not Found dialog box appears on your
screen.
11) Click Yes.

" Wait until the camera driver has been installed.
56
12) Click on the Finish button.

" The Digital Signature Not Found MS Windows dialog box(es)
appear on your screen. Confirm the message as often as neces-
sary until the dialog boxes disappear.
Troubleshooting
13) Open the Windows Device Manager via Start > Settings > Control Pan-
el > System > Hardware > Device Manager.... The Device Manager di-
alog box contains the correct SIS View FireWire camera driver entry:
SoftHard Technology MV4.0 Camera.
57
The Windows Device Manager shows the IEEE 1394 FireWire board and the SIS View FireWire camera driver Soft-
Hard Technology MV4.0 Camera.
14) Install the image-analysis software. During the installation check the
Firewire check box to in the Input Device (Digitizer) Selection dialog
box. Select the ColorView II (Firewire) check box in the following Cam-
era Selection dialog box.
ColorView II • Installation Procedure - Windows XP
Installation Procedure - Windows XP

Installation of the FireWire board IEEE-1394
When is this necessary? This procedure is necessary only if the PC does not have a FireWireTM
interface on-board hardware configuration or if it is not yet equipped with a
FireWireTM board.
the FireWire board
How to install the 1) Turn off the PC and disconnect the power supply.
FireWire board 2) Remove the cover from the PC.
3) One PCI slot is required for the FireWire board. Insert the board into the
slot.
FireWire board installed
58
4) Replace the cover onto the PC.

5) You are now ready to first install the image-analysis software and then
the camera.

Warning Install the image-analysis software first before connecting the camera to the
FireWireTM interface.
System requirements The Color View II camera requires the 3.2 image-analysis software and
Microsoft Windows XP (or higher). The image-analysis software also sup-
ports multithreading on multiprocessors, thus making full use of the power
of the latest multi-CPU PC’s.
camera driver installation Installation of the SIS View FireWireTM camera driver is simply done via the
image-analysis software setup. You are required to select FireWire and
specify the additional camera driver during setup. Then you have to get Win-
dows to use the SIS View FireWire camera driver.
Installation of the image- For installation, follow the instructions given on the monitor.
analysis software
In the image-analysis soft-
ware setup wizard select the
FireWire digitizer and select
your camera type. Select
the Firewire check box in the
Input Device (Digitizer)
Selection dialog box.
59
Related Topics
Installation of the camera driver, S. 62
Then select the Soft Imag-

ing System manufacturer
entry and the Color View II
camera entry.
Note The image-analysis software can support several same or different types of
FireWire cameras simultaneously. You differentiate Color View II cameras
via their serial numbers when creating a new logical input channel via Image
> Set Input... > New Channel. A camera’s serial number is located directly
below the FireWire jack on the camera housing.
The camera’s serial number
is located below the
FireWire jack.
60
How to install the camera 1) The diagram shows a typical installation of a light microscope. You
should have all required components ready at hand.
61
2) Remove the protective cap from the camera.
3) Firmly screw the TV adaptor (1/1,8" or bigger) onto the camera. This
should be done with the CCD-chip side of the camera facing down to
minimize the chance of dust landing on its surface.
4) Install this assembly onto the microscope's camera mount. Some mi-
croscopes may require additional optical elements installed between
the camera mount and the C-mount adapter. You may also need to
align the orientation of the microscope with that of the stage, and also
match the focus of the camera image with that of the eyepiece image.
These settings can be done later when you are able to obtain an image
on the screen.
5) Plug one end of the FireWire cable into the camera connector.
" The FireWire cable plug is not symmetrical. You must make sure
that the cable plug is facing the same way as the camera socket
when you plug the cable into the camera jack. The camera chip
may be ruined otherwise.
Make sure the cable plug is

facing the right way before
you plug it into the camera
socket
6) Plug the other end into one of the small connector sockets on the
FireWire board.
" There are two or more jacks depending on the FireWire board. You
can use any of them.
7) Continue with the camera driver installation before starting the image-
analysis software.
Installation of the camera driver

The SIS View FireWire camera driver is not signed and therefore not used
automatically under Windows XP. After having installed the image-analysis
software and connected the camera to the computer - or vice versa - you
have to go through the following steps to get Windows XP to use the SIS
View camera driver:
1) Open the Windows Device Manager via Start > My Computer > (right
mouse) > Properties > Hardware > Device Manager....
62
" The Device Manager dialog box shows the Generic 1394 Desktop
Camera entry in the Imaging devices section. The exclamation
mark indicates a device conflict.
2) Select the Generic 1394 Desktop Camera entry and click on the Up-
date driver button in the device manager button bar.
3) Select the Install from a list or specific location (advanced user) option
and click on the Next button.
63
4) Select the Don’t search. I will choose the driver to install. option and
click on the Next button.
" The Model list contains the SoftHard Technology MV4.0 Camera
entry.
" If this entry is missing, click on the Cancel button. Repeat the instal-
lation of the image-analysis software including FireWire input
device and Color View II camera selection.
" The next Hardware Update Wizard dialog box will be opened in the
background displaying the message to wait while the Wizard
installs the software.
" Additionally, the Hardware Installation dialog box will pop up in the
foreground.
64
6) Click on the Continue Anyway button.

" The Hardware Update Wizard dialog box which had been in the
background, will become visible.
Related Topics
Installation of the image-analysis software, S. 59
" After having finished the SIS View FireWire camera driver installa-
tion the following dialog box will be opened.
65
7) Click on the Finish button to end the hardware update wizard.

" In the Device Manager dialog box there is still the same Generic
1394 Desktop Camera entry but the exclamation mark indicating
the device conflict should have disappeared. The camera is now
ready for use.
ColorView II • When encountering problems with the camera...
When encountering problems with the camera...

Should you have questions or should problems arise which you cannot solve
with the help of the documentation provided, please feel free to contact our
customer service - preferably via e-mail. Our customer-service personnel
will be more than happy to assist you.
1) Please clarify before hand when and under what circumstances the
problem arose.
• It would be best if you can clearly reproduce the problem/error. You
will therefore make it easier for you and our customer-service per-
sonnel to solve the problem.
66
2) Please write down the exact wording of the (error) messages which ap-
pear.
3) As an alternative, you can make "Snapshots" of these messages. You
can make a snapshot of your active window by using the key combina-
tions [Alt+Print] respectively [Alt+Print]. A copy of the active window is
stored to your clipboard and can be easily attached to an e-mail using
[Strg+V] respectively [Ctrl+V].
4) Since very large e-mails can create transmission problems both for the
sender and the receiver, do not copy "screenshots" of the entire con-
tents of the screen. For the same reasons it is advisable to reduce the
color depth of the snapshots.
5) Should the camera basically work, but problems with image quality
arise, save an image in which the problems are clearly visible in TIF for-
mat. Insert this example image in your e-mail to our customer service.
6) Start the image-analysis software.
7) Open the Info dialog box using the ?>Info... command.
" The Info-Dialog box shows you information about the software
package (Docu, Auto, Pro), die Build-Number and the serial num-
When encountering problems with the camera...
ber of your analySIS-Version as well as the operating system being

used.
• The software data mentioned should be at hand when contacting
our customer service personnel.
8) Please write an e-mail clearly describing the problem (including snap-
shots and example images), as well as the software data from the Info
dialog box to our customer service address:
support.osis@olympus-sis.com
• The quickest and most convenient way is to use the automatic e-

mail settings for your customer service query: by using the ? > In-
fo... > System Info... > Send button, an automatic e-mail is created
which you can then complete and send to us. If you have made an
example in TIF format, please attach this to the e-mail as well. Be-
fore sending, read the notes in your e-mail box the data about your
system which you will send to us in this e-mail.
Should you not be able to send e-mails from your computer, use the
? > Info... > System Info... > Info save button to save the files and
to send them from another computer.
• You may of course contact us via telephone or fax:
Tel.: (+ 49) 2 51 / 7 98 00-444
Fax: (+ 49) 2 51 / 7 98 00-6060
67
CE DECLARATION OF CONFORMITY
Product Type CCD Camera

Brand Name Soft Imaging System
Model Designation Color View II
Name and Address of manufacturer or representative
Manufacturer Soft Imaging System GmbH

Johann-Krane-Weg 39
D-48147 Münster
The product complies with the requirements of the following European directives:
89/336/EWG Directive of the council on the approximation of the laws of Member

States relating to electromagnetic compatibility changed by directive
91/263/ECC, 92/31/ECC and 93/68/ECC of the council.
Compliance was proved by the application of the following
electromagnetic compatibility standards:
EN 55022 : 1998+A12000, cl. B
EN 55024 : 1998
EN 50081-1 : 1992
EN 50082-1 : 1997
EN 61000-4-2
EN 61000-4-3
EN 61000-4-4
EN 61000-4-6
ENV 50204
Date: April 30th, 2002
Dr Manfred Schierjott April 30th, 2002

Date
Index
Display 34
A Display tab
Acquisition, command 11 Image scaling 37
Activate module 25 Over exposure 34
analySIS, version 2 Palette 37
Automatic Gain Display DLL version, ColorView II information 33
Camera Control 15 Duplicate Channels 31
Input 34
E
B EFI
Binning 38 Activate module 25
Black & white mode 27 Exposure time
Gamma Correction 28 Camera Control 12
Set Input 30 Input 32
Black balance 16 intX 20
Exposure time, intX 21
C
F
Camera configuration 29
Camera Control 12 Firmware Version 33
Black balance 16 Fluorescent microscopy 15
Color Settings 13 Black balance 16
Exposure time 12 intelligent Exposure 22
Gain display 15
Partial Readout 18
Sharpen filter 14 G
Sharpness Monitor 17 Gamma Correction
Camera resolution Black & white mode 28
In input channel 32 RGB mode 13
in intX 22
69
Camera Type 33
Clipping 38
Color Settings 13
H
Gamma Correction 13 Housing temperature 33
Saturation 14
ColorView II Black & white mode 27
ColorView II info 33 I
ColorView II specifications 1
Image scaling, Display tab 37
Configure Input 31
intelligent Exposure 11, 19
Display tab 34
intX 19
Format tab 38
Calibration 20
Input tab 31
Exposure time 20
Exposure time correction 21
Resolution 22
D White balance 21
Device ID 33
Device Information 33
Index
K R
Köhler Illumination, Microscope 41 Real time FFT 36
Requirements, PC 2
Resolution
L Input 32
intX 22
Live Acquisition 11 RTFFT Control 29
intX 21
Live overlay 12, 36
S
M Select device 31
Serial Number 66
Microscope 40 Serial number 33
Controlling illumination intensity 3 Set Input 30
Empty Magnification 42 Sharpen filter 14
Illumination 40 Sharpen filter Activate 32
Numerical Aperture 42 Sharpness Monitor 17
Objective 42 Snapshot
Total Magnification 42 Command 19
Useful Magnification 42 intX 21
Microscope Illumination Intensity 3 Special 4
Microscope Settings 3, 40 System Info 67
Mirror 32 System requirements 2
Monitor Settings 4
Monitor's Color Temperature 4
T
N Tab Format 38
Binning 38
NetCam 23 Clipping 38
in the internet 24 Image format 38
70
In the intranet 24 Tab Input 31

Settings 25 Automatic gain display 34
New Channel 30 Exposure time 32
Info 33
Mirror 32
O Resolution 32
Online histogram 35 Sharpen filter 32
Online shading correction 5 Temperature 33
Over exposure 34 Total magnification, microscope 3
Over Exposure Warning 34
U
P Useful magnification, Microscope 42
Partial Readout 18
PC, system requirements 2
Index
W
Web Server 23
White balance 3
Execute 4
General remarks 3
White balance, intX 21
71

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analySIS

© Olympus Soft Imaging Solutions GmbH

ColorView II - Camera - technical specifica-

Camera and light microscope 3

WHITE BALANCE .....................................................................3

Online shading correction 5

OPERATION OF THE SHADING CORRECTION ....................6

ACQUISITION METHODS ......................................................11

Exposure Time ....................................................... 12

INSTALLING THE WEB SERVER .......................................... 23

Usage in Black & White mode 27

Gamma Correction ............................................................ 28

SOFTWARE CAMERA CONTROL ......................................... 30

Appendix I: Microscope Settings 40

Appendix II: Camera Installation 43

INSTALLATION PROCEDURE - WINDOWS 2000 ................ 45

ColorView II - Camera - technical specifications

Modern cameras with the respective software offer numerous possibilities

dividual acquisition methods (live-acquisition, single image, intelligent expo-

Camera and light microscope

Monitor Settings for White Balance

Executing White Balance

3) Click the Set ROI for White Balance button.

Online shading correction 5

Operation of the Shading Correction

Acquiring Correction Images

10) Follow the wizard's instructions to acquire the reference images.

Enabling Shading Correction

Repeat or Update the Reference Acquisitions

Use of the Online Shading Correction

intX and Online Shading Correction

in Black and White Mode".

already with the live acquisition.

Automatic Gain Display and Scaling

black balance is to be used predominately for fluorescence acquisitions in

If you start a live acquisition with an activated Sharpness Monitor or activate

3) Use the Camera Control command.

calculated out of the complete image data in the computer; an acceleration

intelligent Exposure largely makes the acquisition procedure automatic. The

intelligent Exposure is measuring this dark current via the acquisition of an

Installing the web server

You can simply adopt the default settings.

NetCam in the intranet

NetCam in the internet

3) Note the entry in the IP address row (192.168.118.175 in example

Usage in Black & White mode

Display and usage of the acquisition with Real time FFT

forms). The Fourier image is continually updated during a live acquisition. It

The Fourier Image of a Cat-

Software Camera Control

Select Device dialog box

The Input tab

The Display tab

Online histogram while op-

The Format tab

Appendix I: Microscope Settings

Tungsten-halogen Tungtsten-halogen lamps comply with their functionality to a conventional

Adjusting the Con- Now the illuminated area is optimally set.

Objective, Total Magnification and Useful Magnification

Appendix II: Camera Installation

1) Check whether your computer system has a FireWire interface on-

The FireWire board IEEE-1394

Installation Procedure - Windows 2000