Towards Quantitative, Low-Cost, Webcam-Based Microscopy for Medical Diagnostics

Marten Darmawan, Amir Faisal, Gea O.F. Parikesit

Microfluidics Laboratory, Department of Engineering Physics, Faculty of Engineering, Gadjah Mada University,
Yogyakarta 55281, Indonesia

marten_darmawan@yahoo.com, amirf415al@yahoo.com, geaofp@yahoo.com

Abstract

Digital web cameras (popularly known as webcams) have recently gained a significant increase of relevance in
the field of optical microscopy, in particular to allow for quick and do-it-yourself methods in developing low-
cost and portable microscopes suitable for medical diagnostic applications in low-resource areas. Unfortunately,
these methods were published without any systematic explanation and quantitative assessment of the imaging
performances. In this paper we reproduce these do-it-yourself methods, discuss the optical considerations that
are relevant for them, and quantitatively compare their imaging performances with a commercial digital
microscope in order to clarify both the advantages and disadvantages of using the webcam-based microscopes
for medical diagnostics purposes.

Keywords: microscopy, webcam, do-it-yourself, quantitative imaging, medical diagnostics.

I. Introduction for them, and quantitatively assess their imaging

The seminal works performed by Hooke
and van Leeuwenhoek [1] have arguably
transformed optical microscopes into standard
scientific instrumentations, but it was the discovery
of digital sensor technologies that has allowed the
microscopes to function as quantitative
measurement tools [2] versatile for various
applications, spanning from biomedical
investigations [3] to engineering research [4].
Pollak and Hutter reported their method [5] for
mounting a digital web camera (popularly known as
‘webcam’) on an optical microscope, effectively
providing a low-cost version of digital microscopy.
Other methods, using cellular phones, further
demonstrate a level of portability that can allow
digital microscopes to work efficiently in low-
resource area [6,7].
Recently some researchers have devised
methods to develop “webcam-based microscopes”,
where webcams were directly turned into digital
microscopes, hence circumventing the requirement
to possess an existing microscopy setup [8,9,10].
Even though these quick and do-it-yourself
methods have successfully demonstrated both low-
cost and portability, these methods were published
without a systematic explanation and quantitative
assessment of the imaging performances. In this
paper1, we reproduce these do-it-yourself methods,
discuss the optical considerations that are relevant
1
A more comprehensive version of this manuscript has been
submitted for publication in the journal of Optical Engineering:
http://spie.org/x867.xml
performances. Further comparisons with
performances of a commercial digital microscope
were also performed in order to clarify the
advantages and disadvantages of using the
“webcam-based microscopes” for medical
diagnostics purposes.
II. Methods
In general, there are three different do-it-yourself
methods that have been published: (i) the lens-less
method, where the lens is simply removed from the
webcam [8]; (ii) the inverted-lens, where the lens is
inverted and repositioned in the webcam [9]; and
(iii) the extra-lens method, where another lens is
added into the webcam system [10]. In this paper
we focus only on the first two methods, since these
are the most straightforward methods that do not
require components other than the ones existing
within the webcam. To reproduce the do-it-yourself
methods, we use a Prolink webcam (type:
PCC8020; sensor: CMOS; 1280 x 1024 pixels;
interface: USB 2.0; maximum frame rate: 30
frames per second; F# of the lens: 2.8; see
http://www.prolink2u.com/new/products/index.php
?cid=102). The Prolink webcam was chosen mainly
due to its low cost (i.e. around US$ 30.00) and
commercial availability at our geographical area.
Below we describe the configurations of the
reproduced do-it-yourself webcam-based
microscopes. Figure 1 shows the webcam in (a) its
original form, (b) after modification using the lens-
less method [8], and (c-d) after further modification
using the inverted-lens method [9].
(a) (b) (c)
(d)
Figure 1. (a) The webcam in its original form; (b)
After removal of lens and its holder; (c) After
repositioning of the lens and enclosing of the
system in a plastic jar; (d) Schematics of the lens
repositioning (not to scale).
The lens-less method is principally similar
to the lens-less Optofluidic Microscope method
reported by Heng et al. [11], where microscopic
objects were imaged directly into an array of digital
sensors, without any optical element in between.
After the webcam’s body has been dismantled, the
lens and its support were removed from the setup.
A thin glass coverslip (thickness of 170 µ m) was
then put on the CMOS sensor, both to cover it from
dust and to place samples directly on top of the
sensor. Meanwhile, adhesive tapes were used to
cover the electronics around the CMOS sensor from
dusts and wet bio-samples. Due to the close
distance between the sensor and the object, the
CMOS pixel arrays will capture the image with
essentially no magnification. The imaging
resolution will therefore depend only on the pixel
size. For the illumination source, we used a LED
lamp that is positioned above the object plane. A
black tube, made from cardboard, was positioned
around the webcam to prevent stray lights from the
ambience to reach our sensor.
Meanwhile, the inverted-lens method is
actually an adaptation from the “reverse macro”
technique used in photography [12], which is
popularly used as a low-cost alternative to produce
images with a large magnification and a short depth
of focus. Figures 1(c-d) illustrate the configuration
used in our reproduction of the inverted-lens
method. In order to obtain a focused image plane at
the sensor, the object plane need to be located 10.36
mm on top of the inverted lens (see f in Figure 1.e),
based on the following parameters: F# (given in the
product specification) is 2.8; D (measured) is 3.7
mm. After the lens was inverted and repositioned,
the webcam system was enclosed in a plastic jar
(from Skippy peanut butter,
http://www.skippypeanutbutter.com/index.aspx),
such that fine adjustments of the object plane
position can be achieved simply by screwing in or
out the threaded lid. For the illumination, we again
use a LED lamp as the illumination source, located
on top of the object.
In order to comprehensively evaluate the
webcam-based microscopes, we also perform
measurements with a commercial digital
microscope. For this, we use a QX5 Computer
Microscope from Digiblue, with 640 x 480 pixels,
where we use the 60x and 200x magnification
together with the bottom illumination source
(http://store.digiblue.com/QX5_Computer_Microsc
ope_for_PC_p/db12013.htm). Note that such a
microscope has been used for scientific research
before, particularly for low-cost gel electrophoresis
analysis of DNA molecules [13].
All these microscopes were tested using
several periodic objects available at our laboratory.
The first object is a 1951 USAF Resolution Target
(Edmund Optics, Barrington, NJ, USA). This
resolution target, commonly used to determine the
resolution of imaging systems, consists of arrays of
periodic lines (i.e. opaque lines on translucent
background) with varying spatial periodicity. The
second test object is a grooved microchannel (width
= 100 µ m, depth = 1 µ m, translucent grooves in
translucent channels), with grooves periodicity of 5,
10, and 15 µ m [14]. This test object is useful for
testing the contrast and resolution obtained
particularly when both the objects and backgrounds
are translucent, which is relevant should the
developed microscopes be used with unstained
biological cells. The digital images were captured
and stored in our laptops, before being analyzed
using the software ImageJ
(http://rsbweb.nih.gov/ij/).
III. Result and Discussion
III.1. Resolution Target
In this test we focus only on the inverted-
lens microscope and the QX5 microscope, as the
lens-less microscope effectively has no optical
magnification (and its resolution is therefore
defined simply by their pixel size). Figure 2
presents the digital images of the 1951 USAF
Resolution Target captured with the inverted-lens
microscope and the QX5 microscope. Comparisons
of these images indicate two important things. First,
the inverted-lens microscope’s field of view is
comparable to the QX5 microscope with a
magnification between 60x and 200x. Second, the
inverted-lens microscope can resolve a minimum
spatial periodicity of 6.2 µ m (corresponding to
Group 7 and Element 3 in the Resolution Test),
which is superior to the resolutions obtained with
the QX5 microscope, 12.4 µ m (200x
magnification). Note that we also performed the
resolution test with QX5 microscope for 10x and
60x magnification, but these results (not shown
here) were inferior to the one obtained with the
200x lens.
resulting illumination is not ideally uniform, as the
light beams coming out of the LED lamp may
possess different directions of propagation. This, in
turn, leads to shading at the microchannel
sidewalls, as can be observed from the intensity
dips shown in Figures 5(a) and 5(b) that occur at
the sidewalls locations.

(a) (b) (a)

Figure 2. Digital images of the 1951 USAF
Resolution Target captured with: (a) the inverted-
less microscope; (b) the QX5 microscope, with
magnifications of 200x.

III.2. Grooved Microchannel

The test reported in this section is similar,
but more challenging, than in the previous section,
since here both the object and the background are
translucent, which would generally result in low- (b)
contrast images. Figure 3 displays the obtained raw
images, indicating the locations of the grooves in
the channels. Meanwhile, Figures 4 and 5 show the
image intensity distributions along the channel
length and the channel width, respectively, and
Figure 6 displays the Fourier transformation of the
images shown in Figure 3.
The data shown in Figures 4 and 6 shows
that the best spatial resolution, quantitatively
represented by a periodicity of the digital image (c)
intensity, was obtained with the inverted-lens Figure 3. The digital raw images obtained with (a)
microscope. Both the lens-less microscope and the the lens-less microscope, (b) the inverted-lens
QX5 microscope fail to show any detectable microscope, and (c) the QX5 microscope (with 60x
periodicity of the grooves (see Figures 4(a) and magnification), for the grooves periodicity of (left)
4(c)), as also shown in the Fourier spectra (see 5 µ m, (middle) 10 µ m, and (right) 15 µ m.
Figures 6(a) and 6(c)). The inverted-lens
microscope, however, only succeeds to resolve the Im a g e inte nsity v a ria tio n a lo ng the cha n ne l le ng th
210
grooves periodicities of 15 µ m and 10 µ m, but
not 5 µ m (see Figures 4(b) and 6(b)). This agrees 205
well with the results in the previous section, where
the minimum spatial periodicity resolvable with the 200

inverted-lens microscope is 6.2 µ m. 195

Figures 4 and 5 indicate that the lowest
image contrast, quantitatively represented by the
Counts

190
intensity differences between the channel and its
surrounding sidewalls, was obtained with the QX5 185

microscope. This caveat was caused, ironically, by

180
the good illumination distribution of the QX5 5 micron
10 micron
microscope, where the light coming out from the 175 15 micron
LED lamp is passed through a diffuser to ensure
uniform illumination intensity across the whole 170
0 20 40 60 80 100 120 140 160 180 200
field of view, while the microscope object is placed Pixels

sufficiently close to the diffuser. As the webcam-

based microscopes do not use such a diffuser, their (a)
 (a)
Im a g e inte nsity v a ria tio n a lo ng the cha nne l leng th
230
Ima ge intensity va ria tio n a lo ng the cha nnel w idth
5micron 210
220 10micron
15micron
200
210
190
200
180
190
Counts

Counts
170
180

160
170

160 150
5micron
10m icron
150 140 15m icron

140 130
0 50 100 150 200 250 300 350 400 450 500 0 20 40 60 80 100 120
Pixels Pixels

(b) (b)
Ima ge intensity va ria tio n a lo ng the cha nnel leng th Im a g e in te n sity v a ria tio n a lo ng th e ch a n ne l w id th
123 126
5micron
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15micron 124
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5micron
115 114 10 micron
15 micron
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0 50 100 150
Pixels 112
0 10 20 30 40 50 60
Pixels

(c)
Figure 4. Image intensity distributions along the (c)
channel length (i.e. parallel to the grooves
periodicity), obtained with (a) the lens-less Figure 5. Image intensity distributions along the
microscope, (b) the inverted-lens microscope, and channel width (i.e. perpendicular to the
(c) the QX5 microscope (with 60x magnification). microchannel sidewalls), obtained with (a) the lens-
less microscope, (b) the inverted-lens microscope,
Im a g e inte nsity v a ria tio n a lo ng the cha nnel w id th and (c) the QX5 microscope (with 60x
190
magnification).

180

170
Counts

160

150
5micron
10micron
15micron
140

130
0 10 20 30 40 50 60
Pixels

(a)
(b)
(c)
Figure 6. The Fourier transformation of the raw
images obtained with (a) the lens-less microscope,
(b) the inverted-lens microscope, and (c) the QX5
microscope (with 60x magnification), for the
grooves periodicity of (left) 5 µ m, (middle) 10
µ m, and (right) 15 µ m.
The intensity dips displayed by Figures
5(a) and 5(b) can also be used to quantitatively
estimate the locations of the sidewalls, and
eventually to also estimate the microchannel width.
Table 1 shows the results of these estimations,
using 1000 similar measurements along lines
perpendicular to the channel sidewalls, in which the
sidewalls locations are estimated by finding the
center-of-mass of the dips [16]. The values in the
table, together with the actual width of the channels
(100 µ m), allow us to further estimate two
parameters of the webcam. First, the CMOS pixel
size in the webcam can be estimated as 3.2
µ m/pixel (i.e. 100 µ m per 31.3 pixels). Second,
the effective magnification obtained by inverting
and repositioning the webcam lens can be estimated
by comparing the dimensions (in pixels) in the
images obtained by the lens-less and the inverted-
lens microscopes; the estimated value of
magnification is 2.6x (i.e. 82.5/31.3).
Table 1. Quantitative estimation of the sidewalls
locations and the microchannel width
IV. Conclusions
Based on the results described above, we
can conclude that our reproduction of the inverted-
lens microscope provides the most optimum
performance. It allows us to obtain the best spatial
resolution (6.2 µ m), which is twice better than the
QX5 microscope (12.4 µ m, at 200x
magnification). Equipped with this good resolution,
the inverted-lens microscope was also able to
resolve translucent objects with a translucent
background, for up to a spatial periodicity of 10
µ m, which could be relevant for life sciences
investigations on unstained biological cells that
further can be support medical diagnostics. There
are, however, two caveats in the inverted-lens
microscope. First, the illumination is not uniform
(yet), but this can be further engineered using a
light diffuser. The second caveat is the optical
aberrations (data not shown in this paper), which is
due to the inferior lens quality compared to the
QX5 microscope. This problem, however, can be
solved using digital image processing [17-18]. Note
that the inverted-lens setup would not allow us to
obtain a Koehler-type illumination [19], mainly
because it only employs a single lens.
The lens-less microscope is the easiest
setup that can be developed, but it involves
essentially no magnification. This can also be
advantageous, however, because it also means that
we obtain no optical aberration. The decision to
choose between a lens-less microscope, an
inverted-lens microscope, or a commercial digital
microscope would therefore depend on the specific
requirements of the applications. Another
consideration would be the cost and time available:
the commercially-available QX5 microscope costs
about US$ 100 and is directly ready to use after
purchase, while the Prolink webcam costs only Rp.
300,000.00 (i.e. around US$ 30,00) but requires
several hours to develop.
In conclusion, this paper reports on the
reproduction of do-it-yourself methods to develop
low-cost digital microscopy, discuss the optical
considerations that are relevant for these methods,
and quantitatively compare their imaging
performances with a commercial digital microscope
in order to clarify both the advantages and
disadvantages of the webcam-based microscopes.
While the comparisons discussed in this paper has
been the result of measurements using a Prolink
webcam-based microscope and a QX5 microscope,
a similar analysis method can obviously be
employed to compare the imaging performances of
any arbitrary pair of a webcam-based microscope
and a commercial digital microscope.
V. Acknowledgement
We gratefully acknowledge discussions
with Alex Gunawan, Aura Mimosa Nugrowati, and
Indraswari Kusumaningtyas.
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