Quantitative measurements of cerebral blood flow with near-infrared spectroscopy

: We propose a new near-infrared spectroscopy (NIRS) method for quantitative measurements of cerebral blood flow (CBF). Because this method uses concepts of coherent hemodynamics spectroscopy (CHS), we identify this new method with the acronym NIRS-CHS. We tested this method on the prefrontal cortex of six healthy human subjects during mean arterial pressure (MAP) transients induced by the rapid deflation of pneumatic thigh cuffs. A comparison of CBF dynamics measured with NIRS-CHS and with diffuse correlation spectroscopy (DCS) showed a good agreement for characteristic times of the CBF transient. We also report absolute measurements of baseline CBF with NIRS-CHS (69 ± 6 ml/100g/min over the six subjects). NIRS-CHS can provide more accurate measurements of CBF with respect to previously reported NIRS surrogates of CBF.


Introduction
Maintaining adequate cerebral blood flow (CBF) is critically important for brain function and tissue viability, especially for patients in the neurocritical care unit (NCCU) with conditions such as acute ischemic stroke, traumatic brain injury, and subarachnoid hemorrhage [1,2]. Although multiple factors may contribute to the poor outcome in those patients, a major focus of the intensive care is to prevent delayed ischemia due to low blood supply to the brain tissue [3]. Continuous bedside monitoring of CBF could improve outcome by providing the ability to detect any signs of impaired CBF in the patients and allow for suitable interventions to prevent ischemia before permanent tissue damage occurs.
Conventional imaging techniques, such as oxygen-15 positron emission tomography (PET), single-photon emission computer tomography (SPECT), perfusion magnetic resonance imaging (MRI), and xenon-computed tomography (Xe-CT), can measure CBF accurately, but these techniques are not suited for clinical bedside monitoring because they are costly, unable to provide continuous monitoring, involve major instrumentation, and may require radiation exposure and patient transport [4]. Transcranial Doppler ultrasound (TCD) is more common for clinical practice because this technique can record temporal changes in cerebral blood flow velocity (CBFV) in large arteries noninvasively and continuously. However, TCD cannot probe microvascular blood flow, which limits this technique from monitoring regional ischemic conditions in the brain. Besides, performance of TCD is operator dependent and may not be applicable in 10-15% of patients due to lack of ultrasound transmission windows [5,6].
Optical techniques using near-infrared light are promising for portable, continuous, longterm and noninvasive monitoring of regional CBF at bedside settings. Hemoglobin difference ([HbD]), defined as the difference between the tissue concentrations of oxy-hemoglobin ([HbO 2 ]) and deoxy-hemoglobin ([Hb]), is a quantity measured by near-infrared spectroscopy (NIRS). [HbD] has been observed to be highly correlated with CBF [7]. However, [HbD] is not solely sensitive to CBF but also to cerebral blood volume (CBV) and cerebral metabolic rate of oxygen (CMRO 2 ). Furthermore, [HbD] does not take into account the delay, due to the blood transit time in the microvasculature, between tissue concentrations of [HbO 2 ] and [Hb] and the driving changes in CBF. Diffuse correlation spectroscopy (DCS) is another optical technique that can measure CBF directly based on the decorrelation of the intensity fluctuations of the multiple-scattered light [8][9][10]. Relative changes in blood flow index (BFI) measured by DCS have been shown to correlate with relative changes in CBF as assessed by standard modalities, such as TCD [11], Xe-CT [12], arterial spin labeling MRI (ASL-MRI) [13], and velocity encoding MRI (VENC-MRI) [14]. However, DCS measurements do not provide absolute CBF, which hampers the capability of DCS to determine when CBF has fallen under critical levels. Recently, we introduced a method, namely coherent hemodynamics spectroscopy (CHS), to characterize cerebral hemodynamics that are coherent with a given physiological quantity and we have described how it may be used to obtain CBF from NIRS measurements [15][16][17][18]. This method is based on an analytical model that relates [HbO 2 ] and [Hb] to perturbations in CBV, CBF, and CMRO 2 [15,19]. CHS improves on the [HbD] estimation of CBF dynamics by correcting for CBV changes and for the blood transit times in the capillary and venous compartments. Quantitative measurements of capillary blood volume and capillary transit time by CHS also provide absolute values of local CBF. The clinical applicability of CHS and its ability to provide absolute CBF at baseline has been reported in our previous work [17,20,21]. Here, we report for the first time dynamic measurements of CBF with NIRS-CHS, their comparison with [HbD] and DCS measurements, and a critical analysis of absolute and relative measurements of CBF with NIRS-CHS.
The scope of this work is to validate the temporal dynamics of microvascular CBF measured with NIRS-CHS in a protocol that involves induced transient changes in mean arterial pressure (MAP). The blood pressure changes were induced by a rapid deflation of a pneumatic cuff applied to both subject's thighs after 2 minutes of sustained inflation at a super-systolic pressure. Thigh cuff deflation is a well-established method for producing transient changes in arterial blood pressure and in CBF measured by TCD [11,22,23]. We have validated the temporal dynamics of CBF measured with NIRS-CHS by comparison with relative CBF measured concurrently with DCS in the same tissue volume. In addition to the validation of dynamic CBF measurements, we also discuss the sensitivity to model parameters and the effect of model assumptions on NIRS-CHS measurements of CBF.

Subjects
Six healthy subjects (3 females; 3 males; age range: 22-33 years) participated in this study. The Tufts University Institutional Review Board approved the experimental protocol, and the subjects provided written informed consent prior to the experiment. Experiments were performed in accordance with the Declaration of Helsinki.

Data acquisition and measurement protocol
Concurrent and co-localized NIRS and DCS measurements were performed. The NIRS instrument is a commercial frequency-domain tissue spectrometer (Imagent, ISS, Inc., Champaign, IL) that operates at wavelengths of 690 and 830 nm and at a modulation frequency of 140 MHz. The DCS instrument is home-built according to the specificiations of previously reported systems [10,13,14]. It uses a long-coherence-length continuous-wave laser (DL785-070-3O, CrystaLaser, Inc., Ren, NV) emitting light at 785 nm, and a bundle of four co-localized few-mode optical fibers (SMF-28-100, NA: 0.14, core: 8.2 μm, single-mode cutoff wavelength at 1260 nm, Thorlabs, Newton, NJ) for collection. The detection fibers were wrapped around a 15-cm diameter cylinder to convert higher-order modes into nonpropagating leaky modes. This setup of detection fiber is similar to what have been reported in literature [24], which results in bending-insensitive transport of six modes at 785 nm for the detection fibers. The detected light for DCS is delivered to an avalanche photodiode if the intensity was too low, i.e. if the detected photon count rate was less than 5 kHz. The method assumes the mean squared displacement of the moving scatterers to follow a Brownian motion model [8]. For the fitting procedure, absorption and scattering coefficients . We restricted the fit to the early portion of the 2 ( ) g τ curve, wherein 2 ( ) g τ was greater than 1.05, to give more weight to longer photon pathlengths and increase the sensitivity of the BFI signal to cerebral contributions [26]. Then, the relative CBF measured by DCS ( rCBF ( ) DCS t ) is computed as follows: where 0 BFI is the average BFI value during baseline. The time traces of rCBF DCS were smoothed via a moving average window size of 3 frames (~3.6 s). Examples of 2 ( ) g τ from one experiment are shown in Fig. 2 during baseline and at 5 s following the cuff release. We note that the steeper slope of 2 ( ) g τ during baseline compared with the one after cuff release is consistent with higher blood flow during baseline. With the setup of the detection fibers, we initially obtained β = 0.15 ≅ 1/6 as expected for six-mode detection fibers. Here in this study, the correlation factor β = 0.12 is lower than the theoretical prediction. We hypothesize that this could be due to laser instability. However, we note that β is constant during the whole experiment, and since we only care for relative changes in CBF measured by DCS we expect that this low β will not affect the results of this study.

Near-infrared spectroscopy
We obtained absolute baseline cerebral concentrations of oxy-hemoglobin ([HbO 2 ] 0 ), deoxyhemoglobin ([Hb] 0 ), and total hemoglobin ([HbT] 0 ) from the initial baseline measurement with the multi-distance optical probe by applying diffusion theory for a semi-infinite homogeneous medium. The method to calculate absolute optical properties from multidistance amplitude (AC) and phase measurements in frequency-domain NIRS is described elsewhere [27]. Optical signal changes collected with the NIRS/DCS probe at a 3.5 cm source-detector distance were translated into relative changes in cerebral hemoglobin concentrations

and [HbT]
Δ by applying the modified Beer-Lambert law [28]. Differential pathlength factors for 690 and 830 nm light were determined from the tissue optical properties measured at baseline [29]. Here, we did not apply corrections that we have previously considered for water contributions to absorption [30] and for the layered structure of the examined tissue [31]. The reason is that these two corrections would modify the cerebral concentrations in opposite directions (water-corrected [HbT] is lower and layercorrected cerebral [HbT] is greater than uncorrected [HbT]) and would still be associated with assumptions (the volume fraction of water in tissue) and approximations (a tissue structure made of regular layers).
AC amplitudes (rather than average DC intensities) at the two wavelengths were used for the conversion of hemoglobin concentrations via the modified Beer-Lambert law. We found that using AC amplitudes for NIRS and the optical fiber arrangement of the NIRS/DCS probe

Hemodynamic model and fitting procedures
The CHS model describes how the time traces of [HbO 2 ], [Hb], and [HbT] are related to the time-dependent changes in cerebral blood volume (CBV), cerebral blood flow (CBF), and cerebral metabolic rate of oxygen (CMRO 2 ) [15,16,19]. The key equations are: where * denotes the convolution operator. Lowercase notation denotes normalized relative changes in CBV, CBF, and CMRO 2 with respect to baseline (i.e.,  [16,19]. The baseline cerebral blood volume is defined as: or: To clarify how the retrieval procedure works, we divide the variables/parameters of Eqs. (2)-(4) into three categories: (a) measured, (b) assumed, and (c) fitted, as described below. , and the specific value used (0.3) was based on reported measurements with positron emission tomography in the living human brain [34].
For the dynamic variables, we assumed cbv (c) (t) = 0 based on the fact that cerebral capillary recruitment and dilation has been reported to be negligible [35][36][37]. From this assumption, cbv(t) is then expressed as the weighted sum of cbv (a) (t) and cbv (v) (t): Recent studies by our group have suggested that cerebral blood volume changes in response to MAP dynamics are dominated by venous contributions in the lower frequency range (f < 0.2 Hz) [38,39]. On the basis of these studies, we partition the total cerebral blood volume changes into 80% venous and 20% arterial. This implies that Note that in Eq. (8) the coefficient ( one of the fitting parameters. Finally, for the inversion procedure, we considered cbf(t) and cbv(t) to be related by a high-pass impulse response function [ ( ) ( ) ] that depends on a cutoff frequency for cerebral autoregulation (f c (AR) ). This relationship assumes that cbf(t) could be estimated from cbv(t) by an RC high-pass filter relationship where k is the inverse of the modified Grubb's exponent [16,33].
, and k. They are obtained by fitting the measured [HbO 2 ](t) and [Hb](t) over the 10 s following the cuff release to Eqs. (2) and (3). The fit was performed using the MATLAB function "fmincon" to minimize the sum of squared differences between the model prediction and the measured data. The parameters were obtained from the fit by setting limits within physiological ranges [16]. We used the MATLAB function "GlobalSearch" that uses a scatter-search mechanism for generating trial points within pre-defined bounds, and finds the best local minimum from those trial points with "fmincon". To compute the error on each parameter, we applied a bootstrapping technique [40]. We computed the residuals as the difference between the data and the best fit. Then the residuals were randomly sampled and added to the fit to represent a new data set. We performed the fit again on the new data set and repeat this sampling procedure 20 times to obtain a distribution of parameter values. The standard deviation of the 20 results was used to represent the error on the fitting parameters. We found that performing the fit by using "GlobalSearch" and bootstrapping give similar results to using 48 different sets of initial guesses evenly spreaded throughout the range of upper and lower bounds of the five parameters.
From the fitted parameters, an absolute value of CBF at baseline (CBF 0 ) is computed as follows in units of milliliters of blood per 100 grams of tissue per minute (ml/100g/min) based on the central volume principle [17]: Where ρ b is the mass density of brain tissue, taken to be 1.04 g/ml. : where the tilde denotes Fourier transformation with angular frequency ω. .
We note that only three out of the five NIRS CHS ω in Eq. (11). Specifically, we released the assumption of an RC high pass filter relationship between cbv(t) and cbf(t).

Statistical analysis
To compare the dynamic responses of [HbD] ([HbO 2 ] -[Hb]), CBF NIRS-CHS , and rCBF DCS , we measured the values of time-to-minimum (t min ) and time-to-half-recovery (t 1/2 ) (in units of second) for these time traces in the interval of 20 s following the thigh cuff deflation. The minimum value following the cuff release defines the maximum decrease from baseline, and the time interval between the cuff release and the time of the minimum is defined as t min . We denote with t 1/2 the time needed to recover from the minimum value to half of the maximal decrease. A two-tailed signed-rank test was performed to determine statistically significant differences between t min and t 1/2 obtained from [HbD], CBF NIRS-CHS , and rCBF DCS . Analyses were performed with MATLAB, and statistical significance was associated with p-values < 0.05.

CBF dynamics with NIRS-CHS
In this section, we study how variations or uncertainties in the CHS parameters and in the relative arterial-to-venous contributions to cerebral blood volume changes affect the amplitude and temporal dynamics of CBF in response to the cuff release. In other words, how the key parameters in Eq. (11) affect the calculation of  -cbf ( ) NIRS CHS ω and its inverse Fourier transform which yields its temporal dynamics. Based on the average NIRS measurements from the subjects reported in this paper, we simulated temporal traces of Δ[HbO 2 ] and Δ[Hb] following the cuff release ( Fig. 3(a) could be generated as shown in Fig. 3     . This is the correction for the delay of Δ[HbD](t) with respect fo cbf(t) due to capillary and venous blood transit times.

(b). cbv(t) is generated as the normalized change in
As relative arterial-to-venous baseline blood volume increases from 0.1/0.5 = 0.2 to 0.5/0.1 = 5, cbf(t) peaks 0.6 s later and decreases 4-10% more. This latter result can be explained by the fact that changes in CBF do not affect the concentrations of Hb and HbO 2 in the arterial compartment [15], so that a greater sensitivity to the arterial compartment translates given changes in [Hb] and [HbO 2 ] into greater changes in CBF.
One common effect that one can observe in Fig. 4 is the greater decrease in cbf(t) when the dynamic blood volume changes are venous (right panels of Fig. 4) vs. when they are arterial (left panels of Fig. 4). This effect could be explained mathematically by Eq. (11) based on a PET study [34], and we fitted for The value of the transit time in the capillaries, t (c)~0 .7 s, is within the reported range of 0.3 to 1.7 s as measured by fluorescence videography in rats [41]. The transit time in the veins, t (v) (which here represents the venous drainage time within the banana-shaped optically probed volume), of ~5 s corresponds to an overall length of ~5 mm for the draining venules in the probed volume and a typical speed of blood flow in the venule of 1 mm/s [42]. The capillary compartment contribution to the total baseline blood volume, ( )  In that previous study, the change in blood pressure was assumed to be equal to the relative change in arterial blood volume at the cardiac pulsation, and the arterial blood volume fraction ( ) 0 0 CBV /CBV a was derived based on this assumption. This procedure led to a much lower value of ( ) 0 0 CBV /CBV a (~0.04) compared to the assumed value in this study. Another difference of that study is that it performed a separate fit for α, which is always multiplied by t (c) in the expressions for capillary and venous saturations [t (c) appears independently of α only in the microvascular impulse functions of Eqs. (2) and (3)].  . Specifically, the time to minimum of CBF from the cuff deflation (t min ) was 5.3 ± 0.7 s with NIRS-CHS, 5.5 ± 0.3 s with DCS and 7.9 ± 0.8 s with Δ[HbD], whereas the time to half recovery of (t 1/2 ) was 9.7 ± 0.9 s for rCBF NIRS CHS − , 9.9 ± 0.8 s for rCBF DCS , and 12 ± 1 s for Δ [HbD]. All values are reported as mean ± standard error over the six subjects. The maximum reduction in CBF NIRS CHS − following the cuff deflation was 10% ± 2%, ranging from 6% to 17%. The corresponding reduction in rCBF DCS was 39% ± 5%, ranging from 20% to 54%. Furthermore, we did not observe a meaningful correlation between the amplitudes of the CBF transients measured with NIRS-CHS and with DCS. We will comment on these results for the amplitude of CBF changes in the discussion section.  We found an average CBF 0 of 69 ± 6 ml/100g/min (mean ± standard deviation) over six subjects, with a range of 53-110 ml/100g/min. Table 2 shows CBF 0 values obtained from the fit for every single-step response in each subject. To estimate the uncertainty of CBF 0 , we assumed relative errors of 5% in 0 [HbT] [44], 10% in ctHb on the basis of healthy ranges in men and women (of course this error may be significantly reduced if ctHb is measured from a blood test on the subject), and 1% in each of t (c) and It is also important to mention a systematic source of error in CBF 0 , which results from the CHS estimate of t (c) . A careful inspection of the CHS equations for [HbO 2 ] and [Hb] [Eqs. (2) and (3)] shows that t (c) almost always appears in conjunction with α in the product αt (c) (α is the rate constant of oxygen diffusion from capillary blood to tissue), through the expressions for <S (c) > and S (v) . The only exception is the appearance of t (c) (separate from α) in the capillary and venous impulse response functions [ (c) ( )

Absolute CBF with NIRS-CHS in human subjects
. Consequenly, the assumed value of α (which is directly related to the baseline CMRO 2 ) has a direct impact on the CHS estimate of t (c) , with greater values of α resulting in smaller values of t (c) .

Discussion
In this study, we applied the mathematical CHS model to obtain dynamic absolute measurements of CBF from NIRS measurements in six healthy human subjects. This is the first time that NIRS-CHS is compared with another optical technique, DCS, that is applied concurrently and in the same tissue volume. We also showed that NIRS-CHS can generate absolute measurements of CBF that are co-localized with measurements of CBV. We observe that the blood flow index obtained from DCS is a direct measure of blood flow and is affected by the speed of moving red blood cells in all vascular compartments, while NIRS-CHS provides an indirect measurement of CBF and is sensitive to the effects of blood flow only in the capillary and venous vascular compartments. Specifically, DCS is based on the time scale of intensity decorrelation, which directly depends on the speed of moving scattering centers (i.e., red blood cells in the blood stream) regardless of the blood compartment. Instead, NIRS-CHS is based on dynamic measurements of [HbO 2 ] and [Hb], which are affected by the oxygen diffusion from blood to tissue and by the speed of blood flow in the microcirculation. A greater blood flow results in a faster intensity decorrelation in DCS and a smaller deoxygenation of hemoglobin in NIRS-CHS (under the assumption of negligible changes in tissue oxygen consumption). Changes of blood flow in the arterial compartment have no effect on the changes in [HbO 2 ] and [Hb] since there is no oxygen exchange in the arteries. On the contrary, changes of blood flow in the capillary compartment will affect the rate of deoxygenation and the concentrations of both HbO 2 and Hb in tissue. The capillary compartment is actively altering the ratio of [HbO 2 ] and [Hb], while the venous compartment is passively propagating those changes. Of course, changes in blood flow likely occur in all vascular compartments, but the NIRS-CHS method is specifically sensitive to capillary flow changes.
In terms of temporal dynamics, we have found that CBF measured with NIRS-CHS is in good agreement with CBF measured with DCS, which is not the case for [HbD] (see Fig. 6). Although NIRS studies in the literature have proposed [HbD] as a surrogate for CBF [45][46][47], validation studies of [HbD] only compared the correlation between amplitude of changes in [HbD] and CBF during perturbations in MAP [7,47]. Instead, the temporal dynamics of [HbD] and CBF were not compared. Equations (2) and (3)  accounts for the time lag between blood flow changes and the associated changes in the cerebral concentrations of HbO 2 and Hb [see the denominator of the righthand-side of Eq. (11)]. This delay results in a shorter time-to-minimum (t min ) and time-to-halfrecovery (t 1/2 ) values in CBF NIRS CHS − as compared to [HbD] (see Fig. 6), and in earlier peaks of cbf ( ) NIRS CHS t − for longer t (c) and t (v) (see Fig. 4 and Table 1). The assumption of negligible changes in CMRO 2 seems reasonable in our protocol, which does not involve brain activation paradigms, and where cerebral hemodynamics are induced by systemic changes in arterial blood pressure. We observe that even during brain activation protocols, the increase of CBF has been reported to be much greater (~30%) than CMRO 2 changes (~5%) [48,49]. We note that when changes in CMRO 2 are not negligible, our approach is still valid but Eq. (11) yields cbf(t) -cmro 2 (t) instead of cbf(t). In this case, NIRS-CHS can be used in conjunction with an independent measurement of cbf(t) to yield cmro 2 (t).
While we found a good agreement between the characteristic times of the rCBF NIRS CHS − and rCBF DCS transients following the thigh cuffs release, their amplitudes of changes were significantly different (about 10% with NIRS-CHS vs. 39% with DCS) and showed no correlation. The reduction in CBF NIRS CHS − of 10%, ranging from 6% to 17%, is in agreement with previous CBF changes measured with TCD, in a similar protocol, which are in the range 10-20% [11,32,50]. This is also in reasonable agreement with the reported changes in rCBF measured with DCS, also in a similar protocol, after correcting for scalp flow contributions (median % decrease of 9.1%, 95% confidence interval of 6.8-11.2%) [11]. These results suggest that the discrepancy in the amplitudes for CBF measured with DCS and NIRS-CHS in this study may reflect different relative sensitivities of the two techniques to blood flow in the scalp and in the brain, which may also result from the different source detector separation used in this study for NIRS (3.5 cm) and DCS (2.5 cm). The unknown blood flow changes in the superficial tissue layers (scalp, skull, etc.) and in deeper brain tissue, and the unknown relative sensitivity to such changes in the NIRS-CHS and DCS techniques, may also account for the lack of correlation between the amplitudes of CBF changes measured with NIRS-CHS and with DCS. The lack of discrimination of extracerebral and cerebral blood flow is a limitation of this study, which will need to be addressed in future work aimed at depth discrimination (through layered models or tomographic reconstruction, or by application of external pressure onto the scalp to suppress scalp hemodynamics [51,52] to identify and correct for extracerebral tissue confounds in NIRS-CHS measurements of CBF. In addition to relative CBF dynamics, we have described how NIRS-CHS can measure absolute CBF and reported an average value of 69 ± 15 ml/100g/min (mean ± betweensubject standard deviation) in six healthy adult human subjects, ranging from 53 to 110 ml/100g/min. In a study of CBF measurement variability, Henriksen et al reported mean values of 50 ± 9, 66 ± 13, and 59 ± 10 ml/100g/min for CBF in the gray matter in 17 healthy young subjects (age range: 20-30 years) as measured with ASL-MRI, diffusion contrast enhanced perfusion (DCE) MRI, and PET, respectively [53]. Grandin et al reported a value of 59 ± 13 ml/100g/min for cortical gray matter blood flow by using MRI bolus tracking and 45 ± 6 ml/100g/min by using PET in 13 healhty male volunteers [54]. Vonken et al reported an average CBF in gray matter of 69 ± 21 ml/100g/min in 41 healthy subjects using dynamic susceptibility contrast MRI [55]. Here, we found comparable or slightly greater CBF values, on average, than reported in literature. It is noted that our absolute CBF measurements using NIRS-CHS depend on various approximations and assumptions in the model, and we need to further study the impact of such assumptions on the accuracy, precision, reliability, and reproducibility of absolute CBF measurements. In particular, we are currently exploring options to minimize the systematic error on t (c) (and therefore on CBF 0 ) associated with the assumed value of the oxygen diffusion from capillary blood to tissue (α). It is possible that these assumptions introduce systematic errors on CBF 0 and would possibly have minimal effects on the reproducibility of CBF 0 measurements on a single subject, so that longitudinal studies or cerebral perfusion monitoring on individual subjects may not be negatively impacted. However, the robustness and reliability of absolute baseline CBF measurements with NIRS-CHS needs to be further demonstrated in future studies.

Conclusion
In this study, we have presented a new optical approach using NIRS in conjunction with CHS to non-invasively and continuously measure CBF in human subjects. We have reported a first comparison of this technique using concurrent and co-localized diffuse correlation spectroscopy (DCS) measurements of local CBF dynamics during transient changes in mean arterial pressure caused by thigh cuffs release following 2 min of cuff inflation at a supersystolic pressure. The CBF dynamics measured with DCS were in much better agreement with CBF NIRS CHS − than with [HbD], a common NIRS surrogate of blood flow. However, we have found a discrepancy in the amplitude of CBF changes between NIRS-CHS and DCS, which we assign to confounding contributions from scalp blood flow and that will be further investigated in future studies. Even though DCS measurements are directly sensitive to the motion of red blood cells, we note that current models of quantification in DCS (i.e. the diffusion correlation equation) rely on NIRS measurements of the temporal trend of absolute optical properties. Therefore, both techniques will benefit from future comparative studies. If NIRS-CHS measurements of CBF will be further validated, they will leverage the typically high signal-to-noise ratio of NIRS, and provide a strong complement to the NIRS assessment of cerebral blood volume. In fact, the continuous co-registration of local CBV and CBF provides a rich hemodynamic characterization that may correlate with functional, physiological, and pathological states.