See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/26268852

High-speed laser Doppler perfusion imaging using an integrating CMOS image

sensor

Article  in  Optics Express · September 2005

DOI: 10.1364/OPEX.13.006416 · Source: PubMed

CITATIONS READS
99 105

2 authors, including:

Theo Lasser
École Polytechnique Fédérale de Lausanne
304 PUBLICATIONS   7,071 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Inflammation: Cellular and vascular aspects View project

Measurement of Diffusion Characteristics in Nanochannels using Nanochannels View project

All content following this page was uploaded by Theo Lasser on 30 December 2014.

The user has requested enhancement of the downloaded file.

 High-speed laser Doppler perfusion imaging
using an integrating CMOS image sensor
Alexandre Serov and Theo Lasser
Laboratoire d'Optique Biomédicale, École Polytechnique Fédérale de Lausanne
CH-1015, Lausanne, Switzerland
alexandre.serov@epfl.ch

Abstract: This paper describes the design and the performance of a new
high-speed laser Doppler imaging system for monitoring blood flow over an
area of tissue. The new imager delivers high-resolution flow images
(256×256 pixels) every 2 to 10 seconds, depending on the number of points
in the acquired time-domain signal (32-512 points). This new imaging
modality utilizes a digital integrating CMOS image sensor to detect
Doppler signals in a plurality of points over the area illuminated by a
divergent laser beam of a uniform intensity profile. The integrating property
of the detector improves the signal-to-noise ratio of the measurements,
which results in high-quality flow images. We made a series of
measurements in vitro to test the performance of the system in terms of
bandwidth, SNR, etc. Subsequently we give some examples of flow-related
images measured on human skin, thus demonstrating the performance of the
imager in vivo. The perspectives for future implementations of the imager
for clinical and physiological applications are discussed.
©2005 Optical Society of America
OCIS codes: (170.1650) Coherence imaging; (170.3340) Laser Doppler velocimetry;
(170.3890) Medical optics instrumentation; (170.4580) Optical diagnostics for medicine

References and Links

1. G.V. Belcaro, U. Hoffman, A. Bollinger, and A.N. Nicolaides, Laser Doppler (Med-Orion Publishing
Company, London, 1994).
2. K. Wårdell, A. Jakobsson and G.E. Nilsson, “Laser Doppler perfusion imaging by dynamic light scattering,”
IEEE Trans. Biomed. Eng. 40, 309-316 (1993).
3. T.J.H. Essex and P.O. Byrne, “A laser Doppler scanner for imaging blood flow in skin,” J. Biomed. Eng. 13,
189-193 (1991).
4. H. Fujii, K. Nohira, Y. Yamamoto, H. Ikawa, and T. Ohura, “Evaluation of blood flow by laser speckle
image sensing. Part 1,” Appl. Opt. 26, 5321-5325 (1987).
5. J.D. Briers, G. Richards, and X.W. He, “Capillary blood flow monitoring using laser speckle contrast
analysis (LASCA),” J. Biomed. Opt. 4, 164-175 (1999).
6. K.R. Forrester, J. Tulip, C. Leonard, C. Stewart, R.C. Bray, “A laser speckle imaging technique for
measuring tissue perfusion,” IEEE Trans. Biomed. Eng. 51, 2074-2084 (2004).
7. J.D. Briers, “Laser Doppler, speckle and related techniques for blood perfusion mapping and imaging,”
Physiol. Meas. 22, R35-R66 (2001).
8. A. Serov, W. Steenbergen, F.F.M. de Mul, “Laser Doppler perfusion imaging with a complimentary metal
oxide semiconductor image sensor,” Opt. Lett. 25, 300-302 (2002).
9. A. Serov, B. Steinacher, T. Lasser, “Full-field laser Doppler perfusion imaging and monitoring with an
intelligent CMOS camera,” Optics Express 13, 3681-3689 (2005),
http://www.opticsexpress.org/abstract.cfm?URI=OPEX-13-10-3681.
10. R.H. Webb and G.W. Hughes, “Detectors for scanning video imagers,” Appl. Opt. 32, 6227-6235 (1993).
11. A.N. Serov, W. Steenbergen and F. de Mul, “Prediction of the photodetector signal generated by Doppler-
induced speckle fluctuations: theory and some validations,” J. Opt. Soc. Am. A, 18, 622-639 (2001).
12. G.W. Anderson, B.D. Guenther, J.A. Hynecek, R.J. Keyes, and A. VanderLugt, “Role of photodetectors in
optical signal processing,” Appl. Opt. 27, 2871-2886 (1988).
13. M.H. White, D.R. Lampe, F.C. Blaha, and I.A. Mack, “Characterization of surface channel CCD image
arrays at low light levels,” IEEE J. Solid-State Circuits SC-9, 1-12 (1974).
14. R. Bonner and R. Nossal, “Model for laser Doppler measurements of blood flow in tissue,” Appl. Opt. 20,
2097-2107 (1981).

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6416
1. Introduction
Anomalous changes in peripheral blood flow are known to be a good indicator of various
health disorders in the human organism. Laser Doppler imaging is a modern optical technique
that allows accessing blood flow changes reliably and non-invasively [1]. The output of the
instrument is a two-dimensional (2D) flow-related (perfusion, speed, concentration) map over
an area of up to 50x50 cm2. The technique is non-invasive because it involves no physical
contact; the risk of infection and additional discomfort is completely avoided. However,
present commercial laser Doppler imagers do not completely fulfill all requirements imposed
on them by clinical applications. They are slow, special skills are required to use them, and
the interpretation of the obtained results is not always objective. Thus, commercial laser
Doppler imagers and the technique itself are mainly used in medical research projects but not
often in clinical practice, despite the tremendous potential of laser Doppler imaging in the
medical field.
Typical approach to improve the performance of these imagers, particularly to increase the
imaging speed, is to parallelize measurements by using 1D or 2D arrays of photodetectors.
Several blood-flow imaging systems have been designed over the last few years. Scanning
laser Doppler imagers [2, 3], imagers based on speckle contrast analysis [4, 5] or laser speckle
imagers (LSI) [6] were reported. A detailed review of these techniques can be found
elsewhere [7]. However, there is still an important way to go for a widespread use in clinics,
as existing instruments are either difficult to handle, slow, or not sufficiently accurate for
measuring fast flow. Our goal was to develop a high-speed full-field laser Doppler imaging
system that also should be reliable, objective, user- and patient-friendly.
To decrease the imaging time, a parallel detection scheme may be employed wherein the
imaging speed increases by a factor proportional to the number of channels working in
parallel. A 2D matrix of photodetectors is a suitable detection device for that purpose.
Regarding photodetectors, there are presently four different technologies to produce
photosensitive matrices: CCD, CID, simple PIN photodiode arrays, and finally CMOS.
However not all of them can be employed for laser Doppler imaging. One limitation is
imposed by the signal sampling frequency requirements. In laser Doppler blood flowmetry the
measured frequencies are typically in the range of 0 to 20 kHz. The frame rate of CCD can
hardly be faster than 1000 fps, which is not sufficient. Fast CCDs have been reported,
however they are too much expensive for our application. CID technology allows for fast sub-
frame rates however the sensitivity of these devices is better in the blue part of the spectrum,
while laser Doppler blood flow measurements require red and near-infrared lasers. Besides
the CID technology is very expensive. Use of an array of conventional PIN photodiodes is not
attractive due to the insufficient packing density of the sensing elements that results in matrix
being of a low resolution. However, the latest evolution of photodiode matrix technology, i.e.
CMOS image sensors, possesses all the advantages for parallel detection of Doppler signals.
These matrices have high resolution, good response in red, moderate response in near
infrared, and they allow for fast sub-frame rates as pixels can be addressed randomly. Further,
this technology is inexpensive and flexible as a result of the integration of photodetectors and
associated electronics on the same chip.
In 2001 Serov et al. [8] have demonstrated the usefulness of the CMOS image sensor
technology using a non-integrating CMOS sensor for measurements of blood flow by means
of laser Doppler technique. Recently, a new generation of such an imager has been designed
and applied to measurements of perfusion on human skin resulting in flow images every 90
seconds for a 256×256 pixel region of interest [9]. Both of these imaging systems were based
on non-integrating CMOS image sensors. In this paper we describe a new laser Doppler
imaging system that employs an integrating CMOS image sensor. The advantage of the
integrating versus the non-integrating detector, particularly for laser Doppler imaging, is
explained in the following section.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6417
2. Integrating vs. non-integrating detectors
There exist two concepts in CMOS image sensor technology for recording the signals
deposited by the photons in the detector: non-integrating and integrating. In non-integrating
detectors the photon flux is continuously converted into an electrical output signal. To obtain
images, the detector array is read instantaneously by means of sequential photoelectrical
scanning. Each pixel detects only the photons that are received during the time the pixel is
sampled:

Ttot
Δt = . (1)
N

Here Ttot is the time to read out all N pixels of the frame (or sub-frame). Thus, during Δt
one pixel detects X photons:

Ptot
X non_int ∝ Δt . (2)
N
Here Ptot is the total optical power received by N photodetectors.
In the integrating detector concept the total deposited energy is integrated via charges
accumulated while the detector is being hit by photons. The charges are accumulated in a
small capacitor, which at the end of the time frame has to be read out. The charge is then
converted into an output signal that is linearly proportional to number of photons that have hit
the detecting pixel. Either each pixel collects photons during the time the other pixels are read
out (rolling shutter mode), or all pixels collect photons during the integration time and they
are read out immediately thereafter (global shutter mode). The maximum integration time is
equal to the time to read N pixels, Tint=Ttot. Therefore, the number of photons detected by one
pixel of an integrating detector array is

Ptot
X int ∝ Ttot . (3)
N
For both systems the signal to noise ratio (SNR) is determined by the number of detected
photons X [10]:

SNR ∝ X . (4)

So the net advantage in the SNR for the integrating image sensor is

SNRint
= N. (5)
SNRnon_int

Above, we compared two imaging systems, one with an integrating detector array and one
with a non-integrating (scanning) detector array. We have assumed equal detector noise for
both imagers, which is not always true. For completeness, the influence of the temporal noise
on the SNR of each imaging system should also be considered.
For both types of sensors the minimum noise floor consists of thermal noise (TN) and shot
noise (SN).

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6418
 4 ⋅ k ⋅ T ⋅ Bn
TN = iTN
2
= , (6a)
R
SN = iSN
2
= 2 ⋅ q ⋅ I ⋅ Bn , (6b)

I = I photo + I dark . (6c)

Here I photo is the average photocurrent and I dark is the average dark current in the
circuit; k is the Boltzmann constant, T is the temperature in degrees Kelvin, Bn is the noise
equivalent bandwidth, R is the load resistance, and q is the charge of an electron. The value of
the load resistance is determined by the upper cutoff frequency required to pass the signal, fs:

1
R= , (7)
2π ⋅ C ⋅ f s

where C is the capacitance of the photodetector. The SNR is then

is2
SNR = . (8a)
2 ⋅ q ⋅ I ⋅ Bn + 8π ⋅ k ⋅ T ⋅ C ⋅ f s ⋅ Bn
2
I photo
i2
s = . (8b)
M
2
Here is is the mean-square value of the signal and M is the average number of speckles
on a photodetector pixel [11].
In respect to the SNR, we first consider the class of non-integrating devices. In general,
the noise bandwidth and the signal bandwidth are not the same. If the upper cutoff frequency
is determined by a single RC time constant, then the signal bandwidth and the noise
bandwidths are respectively

1
fs = . (9a)
2π ⋅ R ⋅ C
1 π
Bn = = fs . (9b)
4 ⋅ R ⋅C 2
Thus for the non-integrating detector the SNR is

is2
SNRnon_int = . (10)
π ⋅ q ⋅ I ⋅ f s + 4π 2 ⋅ k ⋅ T ⋅ C ⋅ f s2

Second, for the integrating detector, the SNR is expressed as before (eq.(8a)) except that
the noise bandwidth is now defined as Bn = 1 ( 2 ⋅ Tint ) , where Tint is the time interval

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6419
between successive readouts of the diodes (the integration time). Therefore, to match the
signal bandwidth the integration time is determined by

1
Bn = = fs . (11)
2 ⋅ Tint

Now we find that the SNR of the integrating detector is

is2 π
SNRint = = SNRnon_int . (12)
2 ⋅ q ⋅ I ⋅ f s + 8π ⋅ k ⋅ T ⋅ C ⋅ f s
2
2

Thus, at the same photocurrent, the SNR of the integrating detector is about a factor of 1.5
better than for the non-integrating device.
Finally, from eqs.(5) and (12) we find that, compared to the non-integrating detector case,
where only one pixel of the image is measured at a time, the SNR of the integrating detector
array can be increased by a factor of up to

SNRint π
= N. (13)
SNRnon_int 2

The above considerations concern the fundamental difference between the detectors,
however the technological features that influence the detector performance should also be
mentioned. One problem encountered in non-integrating detectors is the dependence of the
time constant on the signal level; this causes the non-integrating detector bandwidth to be
dependent on the signal level. This problem could be eliminated in principle, but at the
expense of an increased noise floor for the associated amplifier circuit [12]. As for the
integrating system, an additional advantage available here is the possibility of reducing the
effect of the thermal noise. This can be achieved by a well-known correlated double sampling
signal processing method [13]. Also, the readout noise of the non-integrating sensor is usually
about an order of magnitude higher than for the integrating one.
Another essential advantage of the integrating detector concept is the flexibility in
selecting the integration time to always match the required signal bandwidth. Since both shot
and thermal noises are distributed over a wide frequency range, reducing the noise bandwidth
effectively reduces the noise in the measurement. Therefore the integration time can be used
as an additional degree of freedom.
3. Experimental configuration; design considerations
Not any CMOS image sensor can be utilized for laser Doppler blood flow measurements. A
limitation arises from the electronic architecture of a particular photosensor matrix. The main
requirement here is a possibility to selectively read out the pixels from a predefined sub-frame
at high-speed. Ideally, the sub-frames would be acquired at frame-rate of up to 40,000 frames
per second as assumed for the maximum sampling frequency in laser Doppler flowmetry.
Another important requirement is the spectral response of the sensor. For laser Doppler blood-
flow measurements the source wavelength should be in the red to near-infrared range. Thus
the spectral response of the detector should be optimized for this range. For our imager a
digital CMOS camera based on the VCA1281 monochrome CMOS image sensor from
Symagery (Canada) was utilized. This sensor operates in rolling shutter mode; it has a 1280H
× 1024V resolution, a 7×7μm2 pixel size, a 40 MHz sampling rate, and an 8-bit ADC. The

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6420
sensor has a specified flat spectral response in the range between 500 and 750 nm. The
camera was connected to the host PC via a fast LVDS (Low-Voltage Differential Signaling)
interface providing for a high-speed transfer of the obtained frames.
For the sample illumination we used a solid-state-diode-pumped laser of 250 mW output
optical power emitting at 671 nm. The laser beam was coupled to a ∅1.5mm plastic optical
fiber. A GRIN (gradient index) lens of ∅1.8mm was placed at the distal end of the fiber. This
configuration produced a uniform illumination of the sample. The illuminated area was up to
∅170mm. The intensity profile of the illuminating beam is shown in Fig. 1b (bottom). A
slight increase of the intensity for higher pixel numbers is caused by the illuminating
geometry – an angle of 9° between the illumination and observation directions. The high-
frequency variations of the intensity are due to speckle effect.
The backscattered light was collected with an f=6 mm objective with an f-number of
f#=1.2. The low f-number objective provided the system with the superior photon collection
efficiency that becomes critical for short integration times (in the range of a few tens of
milliseconds). The imager head was installed on the articulating arm system for providing an
easy access to the object of interest, as shown in Fig. 1. Typically, the imager head was placed
at a distance of 150-250 mm from the measured surface.

RAM (memory)
Laser ADC
CMOS image
sensor Controller
I/O interface
Objective CPU
lens

I/O

Sample
HDD Display

250

200
pixel value, a.u.

150

100

50

0
0 100 200 300 400 500
pixel number

(a) (b)
Fig. 1. a) High-speed laser Doppler imaging system. The imager head was mounted on an
articulating arm to simplify access to the measured objects. b) Block diagram of the laser
Doppler imaging system modules (top); and the intensity profile of the illuminating beam
(bottom).

In general, the geometry of the illuminating beam and the objective lens with respect to
the sample influences the Doppler beat frequency response of the LDI system. For our system
design we only found a minor influence (standard deviation less than 15%) for the imager
head position within ±30° relatively to the vertical detection position. Typically, in optically
dense biological tissues, such as skin, scattered photons loose their initial directions due to

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6421
diffusion rapidly. Finally, the scattering angles as well as the directions of moving blood cells
are random resulting in a less sensitive system response due to variations of the imaging
geometry.
For the imager we have developed software with a simple and functional interface to
control the system. The software allows for changing the sensor parameters, for control of the
data acquisition mode, for acquisition of the data, and for display of the flow-related
(perfusion, concentration, speed) maps. A photographic image of the sample and flow related
maps displayed on the monitor are obtained with the same image sensor; therefore the
obtained flow-maps can be easily associated with an area of interest on the sample. The flow-
map data can be stored as separate images or as a movie. The imager can run in different
modes: i) simple imaging mode, recording a single flow-map image, and ii) continuous
imaging mode, with successive recording of flow-map images.
4. Data acquisition and signal processing
The signal sampling frequency is inversely proportional to the time to acquire one sub-frame.
The sub-frame sampling rate of the sensor depends on its size and the pixel clock frequency.
The clock frequency was fixed at 40 MHz for optimum performance speed/quality; the higher
pixel-sampling rate increases the noise level. The size of the sampled sub-frame finally
defines the signal sampling frequency of the imager. For 256×4 pixels sub-frame the frame
sampling frequency was 30kHz, 256×6 pixels – 20 kHz, 256×8 pixels – 14 kHz, etc.
To obtain one flow map over a region of interest (ROI), which in our case was 256×256
pixels, the ROI must be subdivided in smaller regions (e.g. into 32 sub-frames of 256×8
pixels) and scanned electronically. From 32 to 512 sampled points were obtained for the
acquired time-domain signal for each pixel of the sub-frame, thus the intensity fluctuation
history was recorded for each pixels of the ROI.
The signal processing comprises the calculation of the zero- (M0) and the first-moment
(M1) of the power density spectrum S(ν) of the intensity fluctuations I(t) for each pixel. The
zero-moment is related to the average concentration, <C>, of moving particles in the sampling
volume. The first moment (flux or perfusion) is proportional to the root-mean-square (rms)
speed of moving particles, Vrms, times the average concentration [14]. The governing
expressions are:

∞
Concentration = C ∝ M 0 = ∫ S (ν )dν , (14a)
0
∞
Perfusion = C Vrms ∝ M 1 = ∫ ν S (ν )dν , (14b)
0

∞ 2

S (ν ) = ∫ I (t ) exp ( −i 2πν t ) dt . (14c)

0

Here the variable ν is the frequency of the intensity fluctuations induced by the Doppler
shifted photons. We calculated the power density spectrum using an FFT algorithm applied to
recorded signal variations at each sampled pixel of the ROI. Noise subtraction is performed
upon the calculated spectra by setting a threshold level on the amplitude of the spectral
components. This filtering is applied to reduce the white noise (e.g. thermal and readout
noises) contribution to the signal. Thereafter the perfusion, concentration and speed maps are
calculated and displayed on computer monitor.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6422
 The signal processing is performed with a general purpose PC CPU – AMD-64-3000+.
The total imaging time (including data acquisition, processing and display) depends on the
number of samples obtained for each pixel and the ROI size. For 256×256 pixel ROI the
imaging time is 2.5 sec for 64 samples, 3.5 sec for 128 samples, 5.5 sec for 256 samples, and
10 sec for 512 samples.
5. Measurements and results
In this section we present the results of the measurements obtained with our new imager.
First, we made some in vitro measurements to characterize the performance of the imaging
system in terms of bandwidth, and the linearity of the imager response to velocity and
concentration changes. Second, we demonstrate the performance of our imager in vivo and
present typical flow-maps obtained by measuring microcirculation in human skin.
5.1. In vitro
To measure the imaging system bandwidth and the response linearity to concentration and
velocity changes, a signal from a sine-modulated light emitting diode (LED) was measured at
one pixel. The LED was connected to an analog output of a synthesized function generator
(Stanford Research System, Model DS345).

6000 12
11
5000 10
(M1/M0)(fs/2), Hz

9
(ACRMS/DC)100%
4000 8
7
3000 6
5
2000 4
3
1000 2
1
0
0 1000 2000 3000 4000 5000 6000 7000 0 1000 2000 3000 4000 5000 6000 7000
Signal frequency, Hz Signal frequency, Hz

a) b)
20
16
18 finger skin
14 forearm skin
16
(ACRMS/DC)100%

14 12
12 10
SNR

10
8
8
6
6
4 4

2 2

0.00 0.05 0.10 0.15 0.20 0.25 40 50 60 70 80 90 100

Input signal amplitude, V Integration time, μs

c) d)
Fig. 2. a) The M1/M0 (velocity) imager response as a function of the measured signal
frequency. b) The √M0 (concentration) imager response as a function of the measured signal
frequency. c) The √M0 (concentration) imager response as a function of the measured signal
amplitude. d) The SNR of the system as a function of the integration time for measurements on
finger and forearm skin; error bars represent standard error for each measured SNR.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6423
 The ratio between AC and DC components of the light output power could be set at
required values changing the offset and the amplitude of the LED driver current. The
frequency of the signal could also be changed. The number of points for FFT was 128. The
signal sampling frequency of the imager (2fs) was around 12 kHz. This corresponds to the
bandwidth from DC to 6000 Hz with 100 Hz frequency resolution. The integration time was
82 μs. Experimental results of these in vitro studies are shown in Fig. 2. Figure 2(a) shows the
speed response of the imager M1/M0 as a function of the input signal frequency. The input
signal of 10% modulation depth, ACrms/DC, was measured for the frequency range from 100
to 6500 Hz. A linear dependence of the M1/M0 imager response is found up to the Nyquist
frequency, which matches well to theoretical expectation. Effectively, the measured (M1/M0)fs
value should be equal to the signal frequency, which is clearly seen from the results. After
approximately 6000 Hz, the decay in the imager response is observed due to the aliasing
effect. It should be noted that the digital image sensors do not usually include antialiasing
circuitry in their design; therefore the aliasing effect is virtually unavoidable in the imager. An
antialiasing filter must be employed before the signal is digitized. It is ineffective to apply a
low pass filter on the digitized signal to eliminate aliasing because the effect occurs prior to
the sampling process. Any aliasing effects would already be stored in the digitized signal and
could not be removed by low pass filtering as the effects typically appear as low frequencies
in the signal.
In Fig. 2(b) the ACRMS/DC response of the imager as a function of the input signal
frequency is shown. The ACRMS/DC value is proportional to the square of the M0 value. The
decay in the √M0 imager response is due to the non-zero integration time of the detectors. This
dependence is very similar to the frequency response of a basic low pass filter RC-circuit with
a time constant defined by eq.(9) (see also eq.(11)). A decay of a factor of 0.5 for an RC-
circuit is typical at the high-frequency cut-off. For integrating sensors, the measured signal
response near the cut-off frequency is even smaller being approximately of 0.7 of its
maximum, see eqs.(9).
In Fig. 2(c) the imaging system √M0 response to the amplitude changes of the input signal
is shown. The input signal frequency was fixed at 3000 Hz. The imager signal amplitude
response shows expected linear dependence. At low amplitudes of the input signal the imager
response demonstrates a nonlinearity caused by noise. The results shown in Fig. 2(d) are
discussed in the next section.
5.2. In vivo
In this section we demonstrate the performance of the imager in vivo. In Fig. 3, flow-related
maps obtained on finger skin of a healthy person are shown. The images were obtained for the
imager settings for the bandwidth from DC to 6000 Hz with 100 Hz resolution; the integration
time is 82 μs. The total imaging time is around 3.5 seconds. A smoothing filter is applied to
the row images: the value of each pixel shown was obtained by averaging the row-values of 8
neighboring pixels. The flow maps (perfusion, concentration, speed) are false-coded with 9
colors. This coding is relative and does not mean that the measured perfusion value coded by,
e.g. red, is equal to the value coded by the red color for concentration or speed. The images
clearly show the difference in speed and concentration distributions measured on the fingers.
The lower value for the concentration signal measured on the nail is caused by the higher
amount of non-Doppler-shifted photons reemitted from the relatively thick statically
scattering nail tissue compared to the thin statically scattering stratum corneum layer of the
skin. The signal measured on the nail shows a higher speed of the moving blood cells in the
under-nail tissue. However, we cannot say here definitely whether this is because of the blood
speed was really higher under the nail, or because the measured values were obtained under
two different conditions (e.g. homodyne detection for the skin and heterodyne detection for
the nail) that could make the signal response be different despite the same speeds of the
objects in both cases. This ambiguity is a common problem for all laser Doppler or laser

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6424
speckle imagers and still needs to be investigated. The black and white photographic image of
the object of interest, see Fig. 3(d), is obtained with the same CMOS camera. This image is
useful for determining the anatomical boundaries associated with the perfusion regions
presented in the blood flow maps.

a) b)

c) d)

Low High
Fig. 3. Flow-related maps obtained with the new imager on finger skin (ROI=256×256 pixels):
a) perfusion map [Low=1500 a.u.; High=3000 a.u.]; b) blood concentration map [Low=150
a.u.; High=300 a.u.]; c) flow speed map [Low=500 a.u.; High=1500 a.u.]; d) image of the
object. The imaging area is 5.5x5.5 cm2. The imaging time is 3.5 seconds in total.

Figure 4 shows the perfusion map images obtained during an artery occlusion experiment.
The imager settings were the same as for the measurements described for Fig. 3 and 4 above.
This example demonstrates the performance of the imager in the continuous imaging mode.
The images were taken subsequently every 3.5 seconds, 3.5 s being the imaging time. The
selected maps are shown in the matrix of 4x3 images to see the perfusion changes before,
during and after the occlusion. As expected, there is a decrease of the perfusion signal during
the occlusion. After the occlusion is released the local perfusion raised above the initial value;
this effect is known as reactive hyperemia, shortly after which the blood flow returned to the
initial state.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6425
 Finally, we demonstrate one of the possible applications of the imager. We measured
perfusion changes in skin after a stimulant cream was applied. This cream contains 1% benzyl
nicotinate. This is a pharmaceutical/cosmetical substance that is used in creams against
arthritis pain and a small amount of this cream on the skin stimulates blood flow within a few
minutes. The effect of the cream is clearly seen on the images shown in Fig. 5. The blood
flow starts steadily increasing approximately 1.5 minutes after the cream was applied. The
increased blood flow in the spot was also followed by reddening of the skin that was observed
by eye. The effect is temporal lasting 1-2 hours.

0s 6s 16 s 22 s

29 s 35 s 38 s 41 s

45 s 48 s 58 s 64 s

Low High
Fig. 4. Artery occlusion experiment recording repeated perfusion images in real-time
(ROI=256×256 pixels). Numbers show time (in seconds) when the images were obtained: 0-6
s, before occlusion; 16 s, occlusion on; 22-29 s, occlusion stopped blood flow; 35 s: occlusion
is released; 38-45 s, post-occlusive hyperemia; 48-64 s, restored perfusion level. The imaging
area is 5.5x5.5 cm2. Low=1500 a.u.; High=3000 a.u.

We measured the SNR of the instrument for measurements on finger and forearm skin.
The SNR was measured and calculated according to SNR = SignalACrms 2
NoiseACrms2
as a
function of different integration times of the sensor. The results are shown in Fig. 2(d). The
noise signal was estimated from a M0 flow-map histogram obtained by imaging a statically
scattering white Teflon object. The signal values were found from the M0 flow-map histogram
obtained by imaging of finger and forearm skin. The SNR is increasing for longer integration
times due to the decrease of the noise bandwidth, Bn, (the integration time could also affect
the signal bandwidth). The SNR for finger skin is approximately 1.5-2 times higher compared
for SNR obtained on forearm skin; finger skin perfusion is known to be higher than that of
forearm skin.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6426
 90 sec 97 sec 110 sec

124 sec 138 sec

152 sec

Low High
Fig. 5. Perfusion images obtained with the high-speed laser Doppler imager (ROI=256×256
pixels). The imaging area is 5.5x5.5 cm2. The effect of the stimulant cream (from Induchem
AG, Switzerland) is the increased blood flow in the area where the cream was applied. The
cream was applied on the skin of the inner side of the forearm. Images show the blood flow
changes trough time: at 90, 97, 110, 124, 138, and 152 seconds after the cream was applied to
the skin. The imaging time is c.a. 3.5 seconds per image. Low=500 a.u.; High=2500 a.u. (the
red bar on the latest perfusion image is caused by an accident artifact during the sensor
readout).

6. Conclusion and outlook

In this paper we described the design and performance of a new high-speed laser Doppler
imaging system based on an integrating CMOS image sensor. The use of a 2D matrix of
integrating photodetectors results in an increased SNR of the system compared to the
analogous imagers based on non-integrating detectors. The bandwidth of the imager with
integrating detectors can be adjusted to the bandwidth of the signal thus increasing the SNR of
the measurement. The use of 2D matrix of integrating photodetectors is particularly important
for the parallel detection modality.
We tested the imager in vitro and in vivo. The imager demonstrated a reliable performance
and fast imaging speed: e.g. for a 256×256 pixel ROI the imaging time was 3.5 seconds for
128 points taken for the FFT. This time is essentially smaller compared to the imaging time of
the scanning imagers. Current commercial systems, mainly due to their system design based
on sequential mechanical scanning, need more than 5 minutes to obtain a flow-image of the
same resolution. Here, we measured the changes of blood flow over an extended area of tissue
of 5.5x5.5 cm2 virtually in real time. In principle, for this imager design, the imaging area can
be up to 15x15cm2 for the illuminating laser power of 250 mW. High-resolution real-time
laser Doppler imaging would provide physicians with additional functional information about
microcirculation.
We submit that the imaging time of future LDI systems will be less than one second
approaching more and more the imaging time of the laser speckle imaging (LSI) system
currently accepted as the fastest (video-rate), e.g. [6]. The LSI systems obtain flow-related
information by measuring the contrast of the image speckles. Effectively, the contrast values

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6427
 measured by LSI are directly proportional to the normalized M0 value that is measured by
laser Doppler with integrating photodetectors. The images shown in Fig. 3 demonstrate a
difference between perfusion (M1) and concentration (M0) maps, the former being of primary
interest and the latter being of secondary interest. Thus the objectiveness of the LSI approach
is not really evident in general case. We submit that laser Doppler imaging provides more
objective information rather that the LSI method since with laser Doppler technique the
concentration and speed signals can be measured independently. In LSI these two signals are
always mixed, and thus it can be hard to attribute an exact cause for the changes in the
contrast signal [11]. Generally, the LSI approach functions more as an indicator of flow rather
than an instrument for quantitative measurements of physiological phenomena. However both
systems have the potential to be used in combination, which may lead to even better overall
performance for both technologies. Fortunately, the performance of both methods can now be
compared objectively by measuring the same sample with a single integrating CMOS image
sensor. This is our future plan.
Considering that the medical doctors would use the imager, our main effort was to develop
a reliable, objective, accurate, user- and patient-friendly high-speed imaging system to
measure blood flow in various biological tissues. From our present results, we anticipate
additional patient comfort resulting from measuring times of less than 5 s. Although the
training and experience will still be needed in order for users to interpret the results, the
measurements themselves could be implemented easily due to a superior instrument design in
combination with simple, interactive software.
The new imaging modality demonstrated the paves for an accurate, inexpensive, and easy
to use diagnosis for a widespread dissemination by laboratories and hospitals. The wide range
of applications is one of the major challenges for a future application of the imager. High-
resolution high-speed laser Doppler perfusion imaging is an innovative technique for
diagnosis and assessing the treatment of diseases such as atherosclerosis, psoriasis, diabetes,
skin cancer, allergies, cardiovascular diseases, skin irritation. The new technique could also
be applied for burn assessment, wound healing, and plastic surgery.
Acknowledgments
This research was sponsored by the Swiss Innovation Promotion Agency (KTI/CTI) under
grant 6187.2 MTS-LS. We also thank Mona Wells for helpful discussions and comments
about the manuscript.

#8073 - $15.00 USD Received 8 July 2005; revised 3 August 2005; accepted 7 August 2005
(C) 2005 OSA 22 August 2005 / Vol. 13, No. 17 / OPTICS EXPRESS 6428

View publication stats