Flow velocity estimation using joint Spectral and

Time domain Optical Coherence Tomography

Maciej Szkulmowski, Anna Szkulmowska, Tomasz Bajraszewski,
Andrzej Kowalczyk, and Maciej Wojtkowski*
Institute of Physics, Nicolaus Copernicus University, ul. Grudziadzka 5, PL87-100 Torun, Poland
*Corresponding author: max@fizyka.umk.pl

Abstract: We propose a modified method of acquisition and analysis of

Spectral Optical Coherence Tomography (SOCT) data to provide
information about flow velocities. The idea behind this method is to acquire
a set of SOCT spectral fringes dependent on time followed by a numerical
analysis using two independent Fourier transformations performed in time
and optical frequency domains. Therefore, we propose calling this method
as joint Spectral and Time domain Optical Coherence Tomography (joint
STdOCT). The flow velocities obtained by joint STdOCT are compared
with the ones obtained by known, phase-resolved SOCT. We observe that
STdOCT estimation is more robust for measurements with low signal to
noise ratio (SNR) as well as in conditions of close-to-limit velocity
measurements. We also demonstrate that velocity measurement performed
with STdOCT method is more sensitive than the one obtained by the phase-
resolved SOCT. The method is applied to biomedical imaging, in particular
to in vivo measurements of retinal blood circulation. The applicability of
STdOCT different measurement modes for in vivo examinations, including
1, 5 and 40 μs of CCD exposure time, is discussed.
©2008 Optical Society of America
OCIS codes: (170.4500) Optical coherence tomography; (170.3880) Medical and biological
imaging; (170.4470) Ophthalmology; (280.2490) Flow diagnostics

References and links

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1. Introduction
Optical Coherence Tomography (OCT) provides cross-sectional images of internal structure
of objects with micrometer resolution, and can be either performed in time [1] or frequency
domain [2]. The latter is performed by analysis of interferometric modulations of light
intensity versus optical wavelengths (spectral fringe signal). In Spectral OCT known also as
Spectral domain OCT (SOCT/SdOCT) spectral intereferometric fringes are registered by a
spectrometer. This modality is particularly useful for ophthalmic examinations since it offers
high speed of more than 20 000 A-scans per second and detection sensitivity of more than
95dB [3-6]. The spectral interferometric fringe signals are collected for each lateral position
of the scanning beam and numerically processed to obtain two-dimensional cross-sectional
images representing the amount of back-reflected light versus depth and lateral positions of
the elements of internal structure of an object.
In addition to morphological imaging, SOCT can provide visualization of physiological
parameters [7-11]. At present the retinal blood flow attracts attention as a potentially
important physical parameter in the functional OCT studies. The measurements of

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bidirectional flow by Spectral OCT have been demonstrated by many groups [8, 12, 13]. In
all of these publications, authors used the phase-resolved technique based on linear
relationship between the phase difference of consecutive spectral fringe signals and the
velocity of the moving sample. This technique is analogous to phase-resolved approach
applied in the OCT methods with the time domain detection [14].
Two main factors deteriorate and even preclude velocity recovery in the phase-resolved
OCT techniques: its vulnerability to low signal-to-noise ratio (SNR) [15] and motion artifacts
causing severe phase instabilities [12, 16]. Both problems frequently occur in OCT
measurements of biomedical objects in vivo. Recently, a novel spectral method has been
proposed in order to minimize the influence of phase instabilities so called resonant Doppler
imaging [17], which determines flow velocities on the intensity basis without the need of
extracting the signal phase. Moreover, this method overcomes a phenomenon of blurring of
interference fringes caused by any sample movement during CCD camera integration time.
Another phase independent method was proposed by Wang, et al., [18]. Since this technique
separates the moving and static components within a sample, only blood perfusion imaging is
possible without any flow velocity estimations. This optical angiography relies on introducing
a constant Doppler frequency to modulate the spatial OCT spectral interferograms what was
initially introduced to Spectral OCT by Yasuno, et al., in 2006 [19].
In this paper we present an alternative method of measuring and processing OCT signals
providing information on the spatial distribution of flow velocities. We propose joint time and
frequency domain detection of interferometric OCT signals. The information about the
velocity is obtained directly – similarly to first velocity estimation techniques in TdOCT [20,
21] – from the time dependent beating frequency due to the Doppler shift between the
reference and the sample light beams. Since the velocity estimation is not based on explicit
phase information extracted from interferometric fringes, the proposed variant of Doppler
SOCT is significantly less sensitive to undesired phase instabilities present in low SNR
conditions. Since it does not require any phase wrapping and averaging procedure, it is
accurate for flows close to the upper limit of measurable velocities. This approach does not
require any modifications in hardware of a standard SOCT instrument. High sensitivity of this
method facilitates flow velocity estimation within the time frame required by the regular OCT
imaging. In many biomedical applications, especially in ophthalmology, there are severe
limitations in optical power, which can be delivered to the sample. In such cases a
preservation of high sensitivity requires fixed value of exposure time of the CCD camera
collecting the spectral fringe data. In this case the multi-shot measurements required in
STdOCT can be balanced by reduction of CCD exposure time. And the same it is possible to
keep sensitivity and the optical power delivered to the object at the same levels like in regular
SOCT imaging. In such case the spectral fringe signals can be first processed and then
superimposed giving the structural reconstruction, while time dependent Fourier
transformation will yield information about flow velocities. Comparing to phase-resolved
techniques our method can operate in conditions of much lower SNR still preserving high
accuracy in the whole velocity range, what is crucial in any quantitative measurements of
biomedical samples in vivo in OCT functional studies.
2. Theory
In Spectral OCT Fourier transformation (FT) of the spectral fringe signal measured by a
single exposure of a CCD camera creates one line of a structural cross-sectional image (A-
scan). In order to asses a velocity of a moving interface most of known methods require at
least two spectral fringes acquired in the same lateral position of a sampling light beam [8] or
almost the same lateral position [15]. The acquired set of spectral interferometric fringes can
be described as a function of wavenumber k and time t according to the following equation:

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 I (k , t ) = I 0 (k )⎜
⎛

⎝
∑ R + R + 2∑
l
l r
l
⎞
Rl Rr cos(2 zl (t ) ⋅ k )⎟ ,
⎠
(1)

where I (k , t ) is the spectral fringe signal, I 0 (k ) is spectral density of the light source, Rl and
Rr denote the reflectivity of the sample and reference mirror, respectively; zl (t ) denotes the
optical path difference between the reference mirror and the l -th interface in the sample,
which is time dependent due to the movement of the reference mirror and/or due to the
displacement of the l -th interface in the sample. The displacement of the interfaces within
the sample is usually caused either by a movement of the entire sample itself or by a motion
of the specific interface z l within the sample. If we assume that both, reference mirror
velocity and velocities in the sample are constant during the acquisition of the spectral
fringes, Eq. (1), then the time-dependent position of the l -th interface zl (t ) can be expressed
as:
zl (t ) = zl + δz (t ) = zl + vl ⋅ t (2)

In this relation, z l is the depth position of the l -th interface at the beginning of data
acquisition and vl is the difference between the velocity of the reference mirror and an axial
component of velocity (parallel to the direction of the probing beam propagation) of the l -th
interface. If the l -th interface moves with velocity Vl at an angle α to the probing beam and
the velocity of the reference mirror is equal to vr this can be expressed as:

vl = Vl cos α − vr (3)
Here velocities directed towards a beam-splitter are regarded positive.
One can rewrite Eq. (1) making use of Eqs. (2) and (3).

I (k , t ) = I 0 (k )⎜ ∑ Rl + Rr + 2∑ Rl Rr cos 2 zl + δz (t ) ⋅ k
⎛
(( ) )⎞⎟ (4)
⎝l l ⎠
⎛
I (k , t ) = I 0 (k )⎜
⎝
∑ R + R + 2∑
l
l r
l
Rl Rr cos(2 zl ⋅ k + ωl ⋅ t )⎟
⎞

⎠
(5)

Although the above equations represent the same interference pattern, they emphasize its
different properties. Phase of the oscillatory component visible in Eq. (4) is a function of
wavenumber and its modulation frequency depends on static position zl of l -th interface
and small additional change of δz , that occurs if the l -th interface is moving. Equation (5)
highlights the time-dependence of the interferometric fringes and shows that signal is
modulated in time with frequency ωl . This beat frequency is caused by a Doppler effect, that
arises for each l -th interface along the time axis. This frequency depends on the velocity vl
and is different for each wavenumber k :
ωl = 2vl k (6)
The phase-resolved methods of velocity estimation enables extracting and using the phase of
the signal Eq. (4) while the joint Spectral and Time domain OCT uses the Fourier
transformation to analyze the time-dependent frequency of the signal Eq. (5).

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2.1 Velocity measurement using phase-resolved SOCT
The idea behind the phase-resolved OCT is to determine the phase difference between points
at the same depth in consecutive A-scans. Knowing the change of position δz of l -th
interface, that arises during the time Δt between two consecutive measurements, the velocity
vl of the l -th interface can be calculated. Since δz is much smaller than zl , the difference
between two consecutive measurements appears as a phase change ΔΦ of interferometric
fringes:
ΔΦ λ ΔΦ .
vl = = (7)
2kΔt 4Δt π
Here ΔΦ is the phase difference between successively recorded depth profiles at the same
location of the probing beam. The time between successive profiles acquisition Δt is
approximately equal to the exposure time of the detector, therefore 1 Δt is the frame rate of
an array detector (or equivalently ‘A-scan’ rate). It is important to ensure that ΔΦ is less than
2π . Since the phase can be unambiguously determined in the range of 2π , and the phase
difference is within the range of 4π a procedure of phase wrapping has to be performed to
transform the phase differences to the range (−π , π ) [12, 13]. In the procedure adapted to
recovery of bidirectional flows the following algorithm is used: if | ΔΦ |< π , the phase shift it
is left as it is, and when π <| ΔΦ |< 2π , the phase shift is replaced by ΔΦ − sign (ΔΦ)2π . The
phase differences ΔΦ from the range (0, π ) are considered positive, while those form
(−π ,0) negative. Usually more than two measurements are used to estimate the phase
differences. Wrapped phase differences ΔΦ are averaged out to increase the sensitivity and
accuracy of the velocity estimation [8, 12, 15].
The upper and lower velocity measurement ranges are limited by maximum and minimum
detectable phase difference, respectively. Hence the maximal flow velocity possible to detect
without ambiguity 2π ( ΔΦ ± max = ±π ) is:
λ
v± max = ± , (8)
4Δt
and the minimal one is:
λ ΔΦ min .
v±min = (9)
4Δt π

The value of minimal velocity defines velocity sensitivity. The minimum phase difference
ΔΦ min is equal to the standard deviation of the estimator of the phase differences σ ΔΦ and is
limited by signal-to-noise ratio (SNR) [15, 22, 23]. Assuming signal amplitude much higher
than noise amplitude SNR>>1 the minimum phase difference can be determined as
ΔΦ min = (SNR ) −1 2 [15]. Vakoc, et al., suggest that retinal blood flow velocities <0.2 mm/s
are out of the scope of the standard phase-resolved OCT method in the case of in vivo
imaging [23].
2.2 Velocity measurement using joint Spectral and Time domain OCT
We propose a direct detection of Doppler frequency from a set of M spectra collected in
time increments Δt at the same transverse location of the probing beam, Eq. (5). As the data
are registered and analyzed in wavenumber and time space simultaneously, we called this
method joint Spectral and Time domain OCT (STdOCT).
In order to explain the idea of the method it is convenient to discuss a simple experiment. We
assume that an objective mirror in standard Michelson interferometer is driven with a

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constant speed. M measurements of spectral interferometric fringes are performed at the
same lateral location (Eq. 5). Collected spectral fringe signals undergo standard SOCT
preprocessing consisting of background removal and rescaling to wavenumber domain [24].
Then spectral fringes are plotted as rows, so the abscissa corresponds to wavenumber and the
ordinate to time ( k − t plane, Fig. 1(a)). A Doppler frequency arising from a movement of l -
th interface is visible as a frequency of the signal along the t -axis while the modulation
frequency along k -axis provides information on location of l -th interface. The two-
dimensional set of spectral fringes is analyzed by Fourier transformations, that can be applied
in two separate ways. First FT can be performed “horizontally” thus it converts the STdOCT
data from wavenumber domain to the depth ( z − t plane, Fig. 1(b)). The second FT acts
“vertically” and converts data from time domain to Doppler frequency, that corresponds to
velocity ( k − ω plane, Fig. 1(c)).

Fig. 1. Joint Spectral and Time domain OCT analysis of a mirror moving with
constant velocity; individual images are linked via one- (1D) and two-dimensional
(2D) Fourier forward and inverse transformations (FT and IFT, respectively). The
amplitudes of Fourier transforms are plotted in log scale on panels (b), (c) and (d);
a. 2D interferogram consisting of 25 spectra recorded in time increments
Δt = 40μs ; b. structural image: reconstruction of the axial mirror position; c.
velocity image: Doppler shift for each k is retrieved; d. combined structural and
velocity image representing the Doppler shift distribution in depth; red rectangle
indicates the resulting point that simultaneously localizes moving mirror and its
velocity; e. single line from structural panel, maximum of the peak correspond to the
axial position of the mirror: 0.14mm; f. single line from velocity panel, maximum of
the peak correspond to the mirror velocity: 14.2 kHz, 0.95mm/s.

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 Using both Fourier transforms, one after another, 2D spectral fringes are converted to
velocity distribution in depth ( z − ω plane, Fig. 1(d)). Note that panels (b), (c), (d) display
only amplitudes of complex valued functions. The top-right panel (Fig. 1(b)) corresponds to
data processed in standard SOCT, where the structure of the object is reconstructed. Standard
SOCT uses the modules of Fourier transforms of data to create structural A-scans and the
phases to calculate the velocities with phase-resolved method. Here, M registered spectra
result in M structural A-scans and only structural information is presented with no velocity
information. Maximal optical path difference between the mirror in the reference arm and the
reflecting interface in the sample arm define imaging range in depth z± max . It is connected
with the sampling interval in wavenumber domain Δk of recorded spectra:
π
z± max = ± (10)
2Δk
The variable z , that encodes the position of the sample is chosen to be positive if the
sampling arm is longer then the reference arm, and negative in the opposite case. The image
of the objective mirror is visible as a single interface which apparently is fixed in time. This
image is doubled due to the fact that registered interferogram is a real-valued function [25]. In
STdOCT as well as in standard SOCT complex conjugation of the image is considered
unwanted, thus not displayed in resulting cross-sectional images. Therefore, in the practical
applications only positive depths are displayed.
The bottom left panel (Fig. 1(c)) corresponds to the one-dimensional distribution of
velocity of moving object with no information about structure of the sample. To increase the
sampling density, zero-padding in time domain is applied. The velocity is recovered from the
Doppler frequency ωl , according to Eq. (6). For each known k the velocity can be calculated
separately. Therefore, this representation of data can be also used to find exact relationship
between wavenumbers and pixels in an array detector, and the same to calibrate the
spectrometer very accurately. In this particular case the velocity of moving mirror is
measured to be 14.25 kHz (0.95 mm/s) for k=7.5·106 m-1, Fig. 1(f). The question of velocity
distribution within the object is trivial in the case of a mirror. If the object is more complex,
magnitude and direction of the movement will be known but there would be no information
about the position of the moving interface. Similar to the phase-resolved OCT, the maximal
value of bidirectional flow velocity v± max is given by the time interval Δt between
consecutive measurements of the spectral fringes:
π
v± max = ± (11)
2kΔt
and for Δt = 40us it becomes v± max = ±5.2mm/s .
The bottom right panel, Fig. 1(d) shows the result of two-dimensional Fourier
transformation of the set of M spectral fringes. Coordinates of displayed signals link
positions of all measured interfaces with corresponding velocities. Each interface zl is
represented by two symmetric points appearing with respect to the zero-path-delay and zero-
velocity. The interpretation of resulting points, shown in Fig. 1(d), is following: the mirror
surface localized at z = 140um (Fig. 1(e)) moves with the velocity of 14.2 kHz (Fig. 1(f)).
The sign of velocity value indicates forward or backward direction. The point localized at
( z, ω )=(-0.14 mm, -14.2 kHz) is its complex conjugate.
The points are in fact fuzzy structures. This is due to the fact that their width along the z axis
depends on the axial resolution and the spread along the ω - axis is caused by the dependence
of the Doppler frequency on wavenumber, ω = 2vk , Eq. (6). A velocity value for each z
position is calculated from Doppler frequency indicated by the point with maximal amplitude.

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2.3 Conditions of reliable velocity measurement for phase-resolved SOCT and joint Spectral
and Time domain OCT – SNR analysis
In order to compare both methods we determine conditions under which each of them fails in
velocity estimation. As phase-resolved SOCT operates on phases and STdOCT on signal
amplitudes, we have to analyze how decreasing SNR affects distributions of measured phases
and measured amplitudes.
Since every registered signal can be considered as a deterministic signal and a random noise,
we assume that our interferometric fringe signal I (k , t ) [Eq. (1)] is a sum of a harmonic
component S and a noise component X .
I (k , t ) ≡ S + X (12)
The harmonic component can be expressed by a real-valued function with a given amplitude
s kt , frequency ω kt and the initial phase set to be zero (random variable and its specific value
are denoted by capital and lowercase letter, respectively):
S = skt cos(ω z k ) . (13)
The noise component in turn, can be expressed as a sum of harmonic components with
random phases Φ n (with uniform distribution) and random amplitudes α n :

∑α cos(Φ ) ,
N
X = n n (14)
n
and its statistical properties can be described by a Gaussian function with a mean value x = 0
and a variance σ kt 2 :
1 ⎡ x2 ⎤
X= exp⎢ − ⎥. (15)
2πσ kt2 ⎣⎢ 2σ kt2 ⎦⎥

The distribution of amplitudes α n is identical for all frequencies and has a mean value equal
to zero and a variance equal to σ α2 . The relation between σ α and σ kt is following:
2
σ α2 = σ kt2 . (16)
N
As SOCT measurements are performed in k − t space (Fig. 1(a)), the phase–resolved method
operates in z − t space (Fig. 1(b)) and STdOCT in z − ω space (Fig. 1(d)), the amplitudes
s kt , s zt , s zω are coupled via Fourier transformations. If the Fourier transformation is defined
to conserve power of the signal (E[I2] = E[Z2], Z = FT(I)), the amplitudes are amplified with
respect to a number of points in Fourier transforms N , M :
2 2
s kt = s zt = sωz , (17)
N NM
while the energy of the noise is preserved and equally distributed among the real and
imaginary part of the transform:

σ kt2 = σ Re
2
+ σ Im
2
≡ 2σ zt2 ≡ 2σ ω2z ≡ 2σ 2 . (18)
In order to describe the relation between the signal amplitude and the distribution of the noise
with the same frequency we introduce a parameter κ:
s
κ = kt , (19)
σα

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and we define SNR as a quantity that describes parameters of standard structural tomograms
( z − t space):
s
SNR = 20 log zt = 20 log κ zt , (20)
σ
and any further comparisons between joint STdOCT and phase-resolved SOCT are based on
above definition of SNR.
Table 1 presents relations between the amplitudes and the variances under condition that
Fourier transformation is scaled to preserve signal energy. These results are similar to
analysis performed by Leitgeb, et al. [26]. Comparing the values of signal-to-noise parameter
after one- and two-dimensional Fourier transformation one can see that κ ωz (STdOCT) is
M 1 / 2 times higher comparing to κ zt (phase-resolved SOCT).

Table 1. Signal amplitude skt and the noise standard deviation σ α after one- and two-
dimensional Fourier transformation; N, M – number of points in the first and the second FT,
respectively.

g g’=|FT1D(g)| g”=|FT2D(g)|

N N ·M
s kt s zt = ·s kt sωz = ·skt
2 2

σα N N
σ= σα σ= σα
2 2
s kt s s 1
κ= κ zt = zt = κ κ ωz = ωz = κ zt
σα σ σ M

The distributions of phase and amplitude of signal G is based on the formalism presented
by Goodman to describe the phase and amplitude distribution of a sum of a constant known
phasor S and a random phasor X [27].
After Goodman, the probability density function for the phase is given by the following
expression:
⎧ e − κ zt 2 2 κ cos Φ ⎡ κ 2 sin 2 Φ ⎤
⎪ + zt exp ⎢− zt ⎥ Ω(κ zt cos Φ ) − π < Φ ≤ π
p (Φ ) = ⎨ 2π 2π ⎢⎣ 2 ⎥⎦ , (21)
⎪
⎩ 0 otherwise
b
where function Ω(b ) =
1
∫ e− y
2
2
dy .
2π −∞
With increasing κ zt the density function becomes more narrow, converging towards a Dirac
delta function centered at Φ=0, whereas when the length of the known phasor s decreases to
zero ( κ zt→ 0), this distribution converges to uniform distribution, Fig. 2. As the phase-
resolved SOCT requires phase subtraction, the width of the final distribution broadens two
times. The broader distribution is, the more random wrapped phase differences are detected,
and in turn the averaged value of phase differences is closer to zero.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6016
 Fig. 2. Phase distributions for various values of κzt parameter.

Quoting from Goodman, the probability density function of the amplitude A of the sum of a
constant phasor and a random phasor is given by a Rician density function:

⎧ a ⎡ 2 2 ⎤ ⎛ ⎞
⎪ exp ⎢− a + s ⎥ I 0 ⎜ as ⎟ a>0
⎪ 2
p (a ) = ⎨ σ ⎢⎣ 2σ 2 ⎥⎦ ⎜⎝ σ 2 ⎟
⎠ , (22)
⎪
⎪
⎩ 0 otherwise

Where I 0 (⋅) is a modified Bessel function of the first kind, zero order. As the length of the
known phasor s increases, the shape of density function p A (a) changes from that of a
Rayleigh density to approximately a Gaussian density with mean equal to s .
Joint STdOCT uses the time dependent modulation of the signal, therefore it is successful,
when the amplitude of signal is higher than the maximal amplitude of noise component. This
occurs when the distributions of signal amplitude ( κ zω ≠ 0 , Eq. (21)) and noise ( κ zω = 0 ) are
separated. The minimal value of κzω, which almost always meets this requirement is κ zω = 7 ,
Fig. 3.

Fig. 3. The probability density functions of amplitude for different value of

parameter κzω. Black and red curve correspond to the distributions of amplitude of
pure noise (κzω=0) and amplitude of signal for critical value of (κzω=7) that assure
correct recovery of velocity in STdOCT.

When κ zω converges to zero, the probability of detecting the correct position of signal
amplitude decrease. Every detection of noise causes indication of random velocity, therefore,

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6017
the distribution of recovered velocity broaden and its mean value converges to the center of
the available velocity range (usually to zero).
To determine critical values of SNR below which STdOCT and the phase-resolved
method give false readings, we performed computer simulations based on provided
theoretical model. In order to reconstruct the process of velocity estimation, multiple signals
( M = 30 ) were generated with respect to the shape of spectrum and the probability density
function of amplitude (Eq. (22)) and phase (Eq. (21)). The magnitude of change in harmonic
component between consecutive signals was set to correspond to 0.35 v max and 0.75 v max .
Both methods operates on exactly the same amount of generated signals. The velocity was
recovered for different SNR and the results are shown in Fig. 4.

Fig. 4. Computer simulations of velocity estimations obtained with STdOCT and

phase-resolved SOCT for two different velocities (0.35vmax and 0.75 vmax – solid
and dashed curve, respectively) under the conditions of decreasing SNR.

Both methods fail for certain SNR, however STdOCT is more robust under low SNR. The
failure appears as an underestimate of retrieved velocity values. In joint STdOCT the critical
SNR above which recovered velocity is reliable does not depend on the magnitude of the set
velocity, whereas phase-resolved SOCT fails earlier for higher velocity.
To explain this effect we analyzed the velocity recovery process in the phase-resolved
method within the entire theoretical range. Four different velocities: 0.05, 0.35, 0.75 and 0.95
of vmax were chosen and the velocity estimation for each of them was performed. The
dependence of velocity reading on proximity to the theoretical limits of velocity are shown in
Fig. 5(a). One can see that there is no significant difference in critical values of SNR for
velocities <0.5 vmax, whereas for higher velocities the critical SNR shifts substantially towards
higher values and for 0.95 vmax it is 25dB higher than for 0.05 vmax. In other words, for a given
value of SNR the phase-resolved method can give correct velocity readings for lower
velocities and underestimated values for higher velocities, for example for 20dB 0.95 vmax is
estimated to be 0.75 vmax, whereas lower velocities are correctly recovered. Figure 5(b) shows
simulated values of velocity retrieved by the phase-resolved method versus the velocities set
in the simulation as real ones. In both cases normalization was applied to display the
velocities in respect to the maximum measurable velocity Vmax. It is visible that for higher
SNRs it is possible to measure correctly higher values of velocity. SNR defines the shape of
phase difference distribution. When the width of distribution becomes broad comparing to
phase range, the wrapping procedure causes that some of the phase differences are found as
positive and the other as negative (Fig. 5(b) right panel). Since the final velocity estimation is
based on calculation using several phase differences, its averaged value decreases to zero for
the phase difference close to ±π . Hence, the decreased value of averaged phase difference
underestimates the value of velocity in the phase resolved OCT.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6018
 Fig. 5. Computer simulations of velocity recovery using phase-resolved method. a.
Retrieved velocity values versus SNR for different assumed values of velocity. b.
Retrieved velocity versus assumed values vreal and corresponding phase difference
distributions for v=0.73vmax.

Basing on performed analysis we can determine the conditions under which joint
STdOCT and phase-resolved SOCT fail in velocity measurement. For the phase-resolved
method the critical SNR, that guarantees reliable velocity detection in whole velocity range is
estimated to be >30 dB (Fig. 5), whereas the corresponding value for STdOCT is >6 dB
(Fig. 4). Additionally, since signal amplitude Azω used in joint STdOCT to retrieve velocity
value depends on number of spectra registered in time ( M ), the critical SNR can be
improved M 1 / 2 times with increasing number of measurements (Table 1.). In phase-resolved
SOCT increasing number of spectra does not improve measurement sensitivity, however it
facilitates detection of mean value of phase difference distribution.
Performed simulations do not take into consideration the washout of interference fringes
[17]. This phenomenon deteriorate SNR, hence in this way it affects velocity recovery.
Because both methods suffer from the blurring of interference fringes in the same degree, the
conducted comparison is still valid.
3. Experiment
We use laboratory high resolution Spectral OCT system comprising a broadband light source
(Broadlighter, Superlum, Δλ = 90nm , central wavelength 840 nm), a fiber Michelson
interferometer with fixed reference mirror and custom designed spectrometer with a volume
phase holographic grating DG (1200 grooves/mm) and an achromatic lens focusing spectrum
on 12-bit CCD line-scan camera (Aviiva M2, Atmel), Fig. 6. The experiments were performed
for three different objects: moving mirror, capillary flow and blood flow in human retina. In
measurements of the velocity of moving mirror, a silver mirror was attached to a piezo-
actuator (Physik Instrumente) and it was driven by a triangular voltage signal. The exact
velocity was calculated at the moment of a linear slope of the driving signal from trajectory
registered by the position sensor mounted inside the actuator. Measurements were performed
with A-scan rate of 40.4 μs.
To investigate flows in scattering media, we used a water solution of Intralipid flowing
through capillaries. Two 700 μm thick glass capillaries with flow in opposite directions were
mounted at the angle of 88 deg to the direction of the probing beam ( z -axis) and stable,
laminar flow was ensured by a medical drip system. The sets of 40 spectra were collected at
the same transversal position of the light beam. The acquisition time was set to 52 μs
including 10 μs dead time needed for stabilization of the position of galvo scanner driven by
the stepwise signal. The optical power of the light illuminating the sample was 3.3 mW.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6019
 Fig. 6. Experimental Spectral OCT system : OI optical isolator, FC fiber coupler, PC
polarization controller, DC dispersion compensator, NDF neutral density filter, X
galvo-scanner, L1 lens, DG volume phase holographic grating, CCD line-scan
camera. Three different objects are measured: moving mirror, capillary flow, blood
flow in human retina.

For all retinal blood flow examination the optical power of light illuminating the cornea
was set to 750 μW. In the case of regular exposure time (40 μs) the velocity recovery is based
on 20 spectra, each recorded with 43 μs of repetition time. In measurements with short
exposure time (5 μs and 1 μs) 40 spectra were collected for each lateral position of the
scanning beam. Despite such a short exposure times, the repetition time was >41 μs due to the
dead time of 40 μs between consecutive measurements.
The velocity estimations in phase-resolved SOCT and joint STdOCT are always based on
exactly the same registered data in all comparative experiments. This guarantees that the
differences in velocity recovery are solely caused by the methods themselves, not by
experimental environment or different amount of processed data.
4. Results and discussion
4.1 Moving mirror
In order to validate provided theoretical analyses we performed an experiment with a moving
mirror as an object. To investigate the relation between the velocity estimation and SNR, the

κ κ
light intensity in objective arm had been reduced by a neutral density filter from 20 dB to -
6 dB (from =10 to =0.5) for two velocities 1.9 mm/s and 3.9 mm/s corresponding to 0.35
and 0.95 of vmax. The sets of 30 spectra were collected and then processed to obtain phase-
resolved and STdOCT velocity estimations. Figures 7(a), 7(b) presents achieved velocity
values, which are displayed together with theoretical results demonstrated in Fig. 4.
In the next step we verified the capability of measuring velocities close to the upper limit.
The objective mirror was driven with different velocities within the whole theoretical range.
The intensity of the light in objective arm was constant and resulted in SNR=17.5 dB in
structural tomogram. A single velocity value was calculated from 18 spectra. Retrieved values
of velocities were marked in Fig. 7(c) together with theoretical prediction.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6020
 Fig. 7. Experimental and theoretical comparison of velocity estimations obtained
with STdOCT and phase-resolved SOCT a, b. velocity readings for fixed velocities
equal to 0,75 vmax and 0.35 vmax, respectively, under the conditions of decreasing
SNR; c. velocity readings in whole theoretical velocity range for fixed
SNR=17.5 dB.

The experimental results are in good agreement with the theoretical model. Although
theoretical ranges for both methods are identical, the phase-resolved SOCT fails earlier than
STdOCT and its correctness depends on the magnitude of measured velocity. STdOCT is able
to detect a velocity for SNR ~30 dB lower than the phase-resolved method. This leads to
conclusion that useful velocity range in phase-resolved SOCT is significantly narrower than
in STdOCT.
4.2 Capillary flow
Two experiments were designed, one to verify the method of STdOCT in case of bidirectional
flow in scattering media and the other to compare with phase-resolved SOCT. Figure 8
presents STdOCT images achieved as individual steps during velocity recovery (section 2.2).

Fig. 8. Bidirectional flow of Intralipid: a. 2D structural tomogram; arrow indicates

the direction of incident light b. 2D velocity map, c. Doppler shift distribution in
depth calculated from the single set of 40 spectra corresponding to the green vertical
line on panel (b). d. distribution of maximal intensity signals retrieved from panel
(c); e. plot of the transverse distribution of velocity corresponding to the green
horizontal line on panel (b).

The structural tomogram of the capillaries and the velocity map that indicates bidirectional
flow are shown in Figs. 8(a), 8(b). Single lines in structural and velocity image are obtained
from a set of 40 A-scans. As a first step in velocity recovery, the procedure of zero-padding to
128 points in time space was applied. The signal underwent 2D Fourier transformation and
formed the Doppler shift distribution in depth as shown in Fig. 8(c). The positions of maximal
intensities for each depth z were detected and points that most likely correspond to noise
( κ zω ≤ 2 ) were removed by thresholding procedure, Fig. 8(d). Images (c) and (d) correspond

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6021
to a single line in 2D velocity map (Fig. 8(b)), on which the values of velocity are encoded
using false colors. Figure 8(e) presents a single 1D velocity distribution along the transversal
direction indicated by green horizontal line on the velocity map. All presented velocity
distributions have parabolic shapes, what implies that measured flow is laminar.
To compare the STdOCT and phase-resolved flow velocity estimation in scattering media,
the flow measurements for different concentrations of Intralipid solution and different flow
rates were performed. Concentration of scattering medium affects signal intensity and its
change yields different characteristics of SNR decrease in depth. The acquisition parameters
of OCT data were unchanged. To take full advantage from recording multiple spectra, we
averaged all single A-scans and they were displayed as a single line in structural tomograms.
The results of both methods of flow estimation are presented in Fig. 9. The experiment was
performed under three different flow conditions. In the first case, Fig. 9(a), the concentration
of the Intralipid solution was chosen in such a way that the SNR changes significantly
between front and back side of the capillary. The flow velocity was set to approximately 0.75
of maximal velocity. Both methods return a parabolic distribution of flow velocity, however
phase-resolved method exhibits a slight asymmetry, which increases with depth. Then the
Intralipid concentration was changed to maximize SNR at the back side of capillary and the
flow velocity was increased to exceed vmax. (Fig. 9(b)) We can observe that both methods
give similar readings to approximately half of the velocity range. For higher velocities phase-
resolved method dramatically underestimates the velocity values, and for vmax returns zero. In
STdOCT velocities beyond the range are wrapped and found as negative values. The
distortions of velocity distributions in phase-resolved method appear when SNR decreases or
when velocities are too high (however still in the theoretically achievable range).

Fig. 9. Velocity measurements of Intralipid flow in capillaries for different flow

rates and concentration of the scattering medium. Each panel consists of 4 images: i.
structural tomogram of the inside of capillary; arrow indicates the direction of light
and green line marks the position of 1D velocity distributions presented on the last
image in the row, ii., iii. velocity maps reconstructed by phase-resolved SOCT and
STdOCT, respectively, iv. comparison of 1D velocity distributions retrieved from
both velocity maps: green – phase-resolved, red - STdOCT; arrow indicates the
direction of light; a. high concentration of Intralipid, flow velocity set to be 0.75
vmax, b. lower concentration of Intralipid, flow velocity slightly higher than vmax, c.
high concentration of Intralipid and flow velocity reaches the upper limit, vmax.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6022
The result of velocity estimation in conditions when both effects occur is shown in Fig. 9(c).
It is evident that for the illuminated side of the capillary, the SNR is sufficient to return
accurate velocity values for both methods. With decreasing SNR and increasing velocity
phase-resolved method starts failing, while STdOCT remains unaffected. These results are in
a good agreement with theoretical predictions and with experiments performed with moving
mirror as an object.
4.3 Retinal blood flow, in vivo
As a final test of joint STdOCT and phase-resolved SOCT capabilities in velocity estimations,
the measurement of blood flow in human retina in vivo was performed. Figure 10(a)
demonstrates cross-sectional image of human retina scanned through the region of optic disc.
Figures 10(b) and 10(c) show two-dimensional maps of the flow velocity distribution
obtained with SOCT and STdOCT, respectively.

Fig. 10. Measurement of blood flow velocity in human retina in the region of optic
disc; a. cross-sectional image of the retina performed in close proximity to optic
disc, b., c. velocity maps based on phase-resolved SOCT and STdOCT methods
respectively.

The velocity distributions inside the vessel indicated by green lines in Fig. 10 are
presented in Fig. 11. Although blood flow in large vessels is evident in both methods, the
quantitative velocity estimations differ significantly. The magnitude of the blood velocity in
the center of the vessel is 1mm/s for the phase-resolved method and 4mm/s for STdOCT. The
recovered shapes of velocity distribution also differ greatly. In the phase-resolved method we
observe distortion in the center of the vessel, what may cause a misinterpretation of nature of
flow.

Fig. 11. Comparison of two velocity estimations: green points indicate phase-
resolved SOCT and the red one STdOCT. Diagrams are based on information
extracted from data presented in Fig. 10 (green lines).
The underestimate in phase-resolved SOCT arises from getting out of the useful velocity
range. Beyond the useful range, velocity estimators became progressively underestimated
with decreasing signal and finally decayed to zero. The useful range in STdOCT is wider, but
of course also limited. Another difference is that in STdOCT decreasing signal does not
influence the accuracy of velocity estimation but only the probability of its detection. It gives
confidence that if signal is distinguished from noise and measured velocity does not exceed

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6023
the upper limit, measured velocity is correct. This feature is especially valuable in biomedical
imaging, where diagnoses are based on measured functional parameters.
4.4 Retinal blood flow imaging with ultra-short CCD exposure time
Joint STdOCT comparing to phase-resolved SOCT requires more data to be collected to
estimate velocity value. This experiment is performed to give the proof of concept of time
effective STdOCT. To compensate longer scanning protocols shorter CCD exposure time is
proposed. We assume that higher sensitivity offered by this technique enables measuring the
velocity without any time extension comparing to regular imaging. The capability of
STdOCT to estimate the flow velocity in human retina with extremely short CCD exposure
time of 5 and 1 μs is presented in Fig. 12.

Fig. 12. Velocity estimation of retinal blood flow with extremely short CCD
exposure time: 5 μs and 1 μs for top and bottom line, respectively; ellipses mark
blood vessels a., d. cross-sectional images of human retina in the area of optic disc;
maps of the velocity distribution: b., e. Phase-resolved SOCT method, c., f. Joint
STdOCT method.

The maps in the top line (5 μs) present spatial distribution of the blood flow velocities only in
the larger vessels. The velocity underestimation in the phase-resolved method results in
vanishing of the middle-size vessels in the flow image. STdOCT results obtained for 1 μs
exposure time enable reconstructing one large and three smaller vessels. The map based on
the phase-resolved method shows only faded velocity image of the large vessel.
Unfortunately, the present state-of-the-art of CCD technique does not allow taking full
advantage of extremely short exposure time because the relatively long dead time of the CCD
camera limits the duty ratio to 0.025. In general in OCT studies, there is a pressing need to
collect more data in examination time acceptable for patients. However, in many cases optical
power delivered to the object has to be limited either by the safety regulations or by the power
limitations of the light sources and/or optical components. Recent developments in CCD and
CMOS technologies probably will be soon completed with ultra-fast line scan cameras. This
experiment shows that, in contrast to the phase-resolved method STdOCT is able to benefit
from these improvements. Phase-resolved SOCT requires higher SNR than it is possible to

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6024
achieve with ultra-short exposure time in CCD technique, whereas STdOCT can still operate
reliably in these conditions.
5. Conclusions
We demonstrate the potential of Joint Spectral and Time domain OCT to estimate flow
velocities accurately. In this approach, SOCT measurements are repeated in one position of
scanning beam to register interferogram, that simultaneously depends on optical frequencies
and time. Intensity modulation along axis of optical frequencies encodes information about
structure and the modulation along time axis contains information on velocity.
We analyze known phase-resolved SOCT method under low SNR conditions. It appears that
retrieved velocities are dramatically underestimated and have tendency to decay to zero. This
strong dependence on SNR is especially adverse in the case of measurements of highly
scattering media, where the contribution of noise increases with depth. This may lead to
considerable corruption of the velocity profile for points located deeper. Another cause of
distortion may occur if the velocity is close to the maximal limit of velocity measurable by
the phase-resolved method. Since velocity estimation in STdOCT is based on Doppler shifts,
it is significantly less vulnerable to both effects and is more reliable for any qualitative and
quantitative analysis as it is demonstrated using the same sets of OCT data. The possibility to
unequivocal assessment of blood circulation in human retina renders STdOCT especially
valuable.
Additionally, STdOCT is more sensitive and it is able to detect a correct value of velocity
for SNR lower at ~30 dB than the phase-resolved method. This unique feature can be used to
compensate longer scanning procedure by shortening CCD exposure time. The proof of
concept of time effective STdOCT is ascertained by presented measurements of blood flow in
human retina in vivo for 5 μs and 1 μs exposure time.
Acknowledgments
This work was supported by EURYI grant/award funded by the European Heads of Research
Councils (EuroHORCs) and the European Science Foundation (ESF). Maciej Wojtkowski
acknowledges additional support of Foundation for Polish Science (Homing project and
EURYI) and Rector of NCU for the scientific grant 504-F. Maciej Szkulmowski
acknowledges support of Polish Ministry of Science, grants for years 2005/2008. Anna
Szkulmowska acknowledges support of Polish Science Foundation FNP’2008 scholarship for
young researchers.

#92569 - $15.00 USD Received 7 Feb 2008; revised 7 Apr 2008; accepted 8 Apr 2008; published 14 Apr 2008
(C) 2008 OSA 28 April 2008 / Vol. 16, No. 9 / OPTICS EXPRESS 6025