Application limits of the scaling relations for Monte Carlo simulations in diffuse optics. Part 1: theory.

Monte Carlo (MC) is a powerful tool to study photon migration in scattering media, yet quite time-consuming to solve inverse problems. To speed up MC-simulations, scaling relations can be applied to an existing initial MC-simulation to generate a new data-set with different optical properties. We named this approach trajectory-based since it uses the knowledge of the detected photon trajectories of the initial MC-simulation, in opposition to the slower photon-based approach, where a novel MC-simulation is rerun with new optical properties. We investigated the convergence and applicability limits of the scaling relations, both related to the likelihood that the sample of trajectories considered is representative also for the new optical properties. For absorption, the scaling relation contains smoothly converging Lambert-Beer factors, whereas for scattering it is the product of two quickly diverging factors, whose ratio, for NIRS cases, can easily reach ten orders of magnitude. We investigated such instability by studying the probability-distribution for the number of scattering events in trajectories of given length. We propose a convergence test of the scattering scaling relation based on the minimum-maximum number of scattering events in recorded trajectories. We also studied the dependence of MC-simulations on optical properties, most critical in inverse problems, finding that scattering derivatives are ascribed to small deviations in the distribution of scattering events from a Poisson distribution. This paper, which can also serve as a tutorial, helps to understand the physics of the scaling relations with the causes of their limitations and devise new strategies to deal with them.

Reliable Fast (20 Hz) Acquisition Rate by a TD fNIRS Device: Brain Resting-State Oscillation Studies.

A high power setup for multichannel time-domain (TD) functional near infrared spectroscopy (fNIRS) measurements with high efficiency detection system was developed. It was fully characterized based on international performance assessment protocols for diffuse optics instruments, showing an improvement of the signal-to-noise ratio (SNR) with respect to previous analogue devices, and allowing acquisition of signals with sampling rate up to 20 Hz and source-detector distance up to 5 cm. A resting-state measurement on the motor cortex of a healthy volunteer was performed with an acquisition rate of 20 Hz at a 4 cm source-detector distance. The power spectrum for the cortical oxy- and deoxyhemoglobin is also provided.

A Compact Multi-Distance DCS and Time Domain NIRS Hybrid System for Hemodynamic and Metabolic Measurements.

In this work, we present a new multi-distance diffuse correlation spectroscopy (DCS) device integrated with a compact state-of-the-art time domain near infrared spectroscopy (TD-NIRS) device. The hybrid DCS and TD-NIRS system allows to retrieve information on blood flow, tissue oxygenation, and oxygen metabolic rate. The DCS device performances were estimated in terms of stability, repeatability, ability in retrieving variations of diffusion coefficient, influence of the tissue optical properties, effect of varying count rates and depth sensitivity. Crosstalk between DCS and TD-NIRS optical signals was also evaluated. Finally, in vivo experiments (venous and arterial cuff occlusions on the arm) were conducted to test the ability of the hybrid system in measuring blood flow variations.

Real-Time Dual-Wavelength Time-Resolved Diffuse Optical Tomography System for Functional Brain Imaging Based on Probe-Hosted Silicon Photomultipliers.

Near-infrared diffuse optical tomography is a non-invasive photonics-based imaging technology suited to functional brain imaging applications. Recent developments have proved that it is possible to build a compact time-domain diffuse optical tomography system based on silicon photomultipliers (SiPM) detectors. The system presented in this paper was equipped with the same eight SiPM probe-hosted detectors, but was upgraded with six injection fibers to shine the sample at several points. Moreover, an automatic switch was included enabling a complete measurement to be performed in less than one second. Further, the system was provided with a dual-wavelength ( 670 n m and 820 n m ) light source to quantify the oxy- and deoxy-hemoglobin concentration evolution in the tissue. This novel system was challenged against a solid phantom experiment, and two in-vivo tests, namely arm occlusion and motor cortex brain activation. The results show that the tomographic system makes it possible to follow the evolution of brain activation over time with a 1 s -resolution.

Cerebral oxygenation and blood flow in normal term infants at rest measured by a hybrid near-infrared device (BabyLux).

BACKGROUND: The BabyLux device is a prototype optical neuro-monitor of cerebral oxygenation and blood flow for neonatology integrating time-resolved reflectance spectroscopy and diffuse correlation spectroscopy. METHODS: Here we report the variability of six consecutive 30 s measurements performed in 27 healthy term infants at rest. Poor data quality excluded four infants. RESULTS: Mean cerebral oxygenation was 59.6 ± 8.0%, with intra-subject standard deviation of 3.4%, that is, coefficient of variation (CV) of 5.7%. The inter-subject CV was 13.5%. Mean blood flow index was 2.7 × 10-8 ± 1.56 × 10-8 (cm2/s), with intra-subject CV of 27% and inter-subject CV of 56%. The variability in blood flow index was not reduced by the use of individual measures of tissue scattering, nor accompanied by a parallel variability in cerebral oxygenation. CONCLUSION: The intra-subject variability for cerebral oxygenation variability was improved compared to spatially resolved spectroscopy devices, while for the blood flow index it was comparable to that of other modalities for estimating cerebral blood flow in newborn infants. Most importantly, the simultaneous measurement of oxygenation and flow allows for interpretation of the high inter-subject variability of cerebral blood flow as being due to error of measurement rather than to physiological instability.

Time Domain Near Infrared Spectroscopy Device for Monitoring Muscle Oxidative Metabolism: Custom Probe and In Vivo Applications.

Measurement of muscle oxidative metabolism is of interest for monitoring the training status in athletes and the rehabilitation process in patients. Time domain near infrared spectroscopy (TD NIRS) is an optical technique that allows the non-invasive measurement of the hemodynamic parameters in muscular tissue: concentrations of oxy- and deoxy-hemoglobin, total hemoglobin content, and tissue oxygen saturation. In this paper, we present a novel TD NIRS medical device for muscle oxidative metabolism. A custom-printed 3D probe, able to host optical elements for signal acquisition from muscle, was develop for TD NIRS in vivo measurements. The system was widely characterized on solid phantoms and during in vivo protocols on healthy subjects. In particular, we tested the in vivo repeatability of the measurements to quantify the error that we can have by repositioning the probe. Furthermore, we considered a series of acquisitions on different muscles that were not yet previously performed with this custom probe: a venous-arterial cuff occlusion of the arm muscle, a cycling exercise, and an isometric contraction of the vastus lateralis.

Fast silicon photomultiplier improves signal harvesting and reduces complexity in time-domain diffuse optics.

We present a proof of concept prototype of a time-domain diffuse optics probe exploiting a fast Silicon PhotoMultiplier (SiPM), featuring a timing resolution better than 80 ps, a fast tail with just 90 ps decay time-constant and a wide active area of 1 mm2. The detector is hosted into the probe and used in direct contact with the sample under investigation, thus providing high harvesting efficiency by exploiting the whole SiPM numerical aperture and also reducing complexity by avoiding the use of cumbersome fiber bundles. Our tests also demonstrate high accuracy and linearity in retrieving the optical properties and suitable contrast and depth sensitivity for detecting localized inhomogeneities. In addition to a strong improvement in both instrumentation cost and size with respect to legacy solutions, the setup performances are comparable to those of state-of-the-art time-domain instrumentation, thus opening a new way to compact, low-cost and high-performance time-resolved devices for diffuse optical imaging and spectroscopy.

Optical properties of developing pip and stone fruit reveal underlying structural changes.

Analyzing the optical properties of fruits represents a powerful approach for non-destructive observations of fruit development. With classical spectroscopy in the visible and near-infrared wavelength ranges, the apparent attenuation of light results from its absorption or scattering. In horticultural applications, frequently, the normalized difference vegetation index (NDVI) is employed to reduce the effects of varying scattering properties on the apparent signal. However, this simple approach appears to be limited. In the laboratory, with time-resolved reflectance spectroscopy, the absorption coefficient, µa , and the reduced scattering coefficient, µs ', can be analyzed separately. In this study, these differentiated optical properties were recorded (540-940 nm), probing fruit tissue from the skin up to 2 cm depth in apple (Malus × domestica 'Elstar') and plum (Prunus domestica 'Tophit plus') harvested four times (65-145 days after full bloom). The µa spectra showed typical peak at 670 nm of the chlorophyll absorption. The µs ' at 670 nm in apple changed by 14.7% (18.2-15.5 cm(-1) ), while in plum differences of 41.5% (8.5-5.0 cm(-1) ) were found. The scattering power, the relative change of µs ', was zero in apple, but enhanced in plum over the fruit development period. This mirrors more isotropic and constant structures in apple compared with plum. For horticultural applications, the larger variability in scattering properties of plum explains the discrepancy between commercially assessed NDVI values or similar indices and the absolute µa values in plum (R < 0.05), while the NDVI approach appeared reasonable in apple (R ≥ 0.80).

Time domain functional NIRS imaging for human brain mapping.

This review is aimed at presenting the state-of-the-art of time domain (TD) functional near-infrared spectroscopy (fNIRS). We first introduce the physical principles, the basics of modeling and data analysis. Basic instrumentation components (light sources, detection techniques, and delivery and collection systems) of a TD fNIRS system are described. A survey of past, existing and next generation TD fNIRS systems used for research and clinical studies is presented. Performance assessment of TD fNIRS systems and standardization issues are also discussed. Main strengths and weakness of TD fNIRS are highlighted, also in comparison with continuous wave (CW) fNIRS. Issues like quantification of the hemodynamic response, penetration depth, depth selectivity, spatial resolution and contrast-to-noise ratio are critically examined, with the help of experimental results performed on phantoms or in vivo. Finally we give an account on the technological developments that would pave the way for a broader use of TD fNIRS in the neuroimaging community.

Forward solvers for photon migration in the presence of highly and totally absorbing objects embedded inside diffusive media.

In this paper, after a critical review of the literature, we present two forward solvers and a new methodology for description of photon migration in the presence of totally absorbing inclusions embedded in diffusive media in both time and CW domains. The first forward solver is a heuristic approach based on a higher order perturbation theory applied to the diffusion equation (DE) [denoted eighth-order perturbation theory (EOPT)]. The second forward solver [denoted eighth-order perturbation theory with the equivalence relation (EOPTER) ] is obtained by combining the EOPT solver with the adoption of the equivalence relation (ER) [J. Biomed. Opt.18, 066014 (2013)]. These forward solvers can possibly overcome some evident limitations of previous approaches like the theory behind the so-called banana-shape regions or exact analytical solutions of the DE in the presence of highly or totally absorbing inclusions. We also propose the ER to reformulate the problem of a totally absorbing inclusion in terms of another inclusion having a finite absorption contrast and a re-scaled volume. For instance, we have shown how this approach can indeed be used to simulate black inclusions with the Born approximation. By means of comparisons with the results of Monte Carlo simulations, we have shown that the EOPTER solver can model totally absorbing inclusions with an error smaller than about 10%, whereas the EOPT solver shows an error smaller than about 20%, showing a performance largely better than that observed with solvers proposed previously.

Detectability of hemodynamic oscillations in cerebral cortex through functional near-infrared spectroscopy: a simulation study.

Significance: We explore the feasibility of using time-domain (TD) and continuous-wave (CW) functional near-infrared spectroscopy (fNIRS) to monitor brain hemodynamic oscillations during resting-state activity in humans, a phenomenon that is of increasing interest in the scientific and medical community and appears to be crucial to advancing the understanding of both healthy and pathological brain functioning. Aim: Our general object is to maximize fNIRS sensitivity to brain resting-state oscillations. More specifically, we aim to define comprehensive guidelines for optimizing main operational parameters in fNIRS measurements [average photon count rate, measurement length, sampling frequency, and source-detector distance (SSD)]. In addition, we compare TD and CW fNIRS performance for the detection and localization of oscillations. Approach: A series of synthetic TD and CW fNIRS signals were generated by exploiting the solution of the diffusion equation for two different geometries of the probed medium: a homogeneous medium and a bilayer medium. Known and periodical perturbations of the concentrations of oxy- and deoxy-hemoglobin were imposed in the medium, determining changes in its optical properties. The homogeneous slab model was used to determine the effect of multiple measurement parameters on fNIRS sensitivity to oscillatory phenomena, and the bilayer model was used to evaluate and compare the abilities of TD and CW fNIRS in detecting and isolating oscillations occurring at different depths. For TD fNIRS, two approaches to enhance depth-selectivity were evaluated: first, a time-windowing of the photon distribution of time-of-flight was performed, and then, the time-dependent mean partial pathlength (TMPP) method was used to retrieve the hemoglobin concentrations in the medium. Results: In the homogeneous medium case, the sensitivity of TD and CW fNIRS to periodical perturbations of the optical properties increases proportionally with the average photon count rate, the measurement length, and the sampling frequency and approximatively with the square of the SSD. In the bilayer medium case, the time-windowing method can detect and correctly localize the presence of oscillatory components in the TD fNIRS signal, even in the presence of very low photon count rates. The TMPP method demonstrates how to correctly retrieve the periodical variation of hemoglobin at different depths from the TD fNIRS signal acquired at a single SSD. For CW fNIRS, measurements taken at typical SSDs used for short-separation channel regression show notable sensitivity to oscillations occurring in the deep layer, challenging the assumptions underlying this correction method when the focus is on analyzing oscillatory phenomena. Conclusions: We demonstrated that the TD fNIRS technique allows for the detection and depth-localization of periodical fluctuations of the hemoglobin concentrations within the probed medium using an acquisition at a single SSD, offering an alternative to multi-distance CW fNIRS setups. Moreover, we offered some valuable guidelines that can assist researchers in defining optimal experimental protocols for fNIRS studies.

Statistics of maximum photon penetration depth in a two-layer diffusive medium.

We present numerical results for the probability density function f(z) and for the mean value of photon maximum penetration depth zmax> in a two-layer diffusive medium. Both time domain and continuous wave regime are considered with several combinations of the optical properties (absorption coefficient, reduced scattering coefficient) of the two layers, and with different geometrical configurations (source detector distance, thickness of the upper layer). Practical considerations on the design of time domain and continuous wave systems are derived. The methods and the results are of interest for many research fields such as biomedical optics and advanced microscopy.

Erratum: Assessment of power spectral density of microvascular hemodynamics in skeletal muscles at very low and low-frequency via near-infrared diffuse optical spectroscopies: erratum.

[This corrects the article on p. 5994 in vol. 14, PMID: 38021143.].

Non-Invasive Monitoring of Microvascular Oxygenation and Reactive Hyperemia using Hybrid, Near-Infrared Diffuse Optical Spectroscopy for Critical Care.

The detection of levels of impairment in microvascular oxygen consumption and reactive hyperemia is vital in critical care. However, there are no practical means for a robust and quantitative evaluation. This paper describes a protocol to evaluate these impairments using a hybrid near-infrared diffuse optical device. The device contains modules for near-infrared time-resolved and diffuse correlation spectroscopies and pulse-oximetry. These modules allow the non-invasive, continuous, and real-time measurement of the absolute, microvascular blood/tissue oxygen saturation (StO2) and the blood flow index (BFI) along with the peripheral arterial oxygen saturation (SpO2). This device uses an integrated, computer-controlled tourniquet system to execute a standardized protocol with optical data acquisition from the brachioradialis muscle. The standardized vascular occlusion test (VOT) takes care of the variations in the occlusion duration and pressure reported in the literature, while the automation minimizes inter-operator differences. The protocol we describe focuses on a 3-min occlusion period but the details described in this paper can readily be adapted to other durations and cuff pressures, as well as other muscles. The inclusion of an extended baseline and post-occlusion recovery period measurement allows the quantification of the baseline values for all the parameters and the blood/tissue deoxygenation rate that corresponds to the metabolic rate of oxygen consumption. Once the cuff is released, we characterize the tissue reoxygenation rate, magnitude, and duration of the hyperemic response in BFI and StO2. These latter parameters correspond to the quantification of the reactive hyperemia, which provides information about the endothelial function. Furthermore, the above-mentioned measurements of the absolute concentration of oxygenated and deoxygenated hemoglobin, BFI, the derived metabolic rate of oxygen consumption, StO2, and SpO2 provide a yet-to-be-explored rich data set that can exhibit disease severity, personalized therapeutics, and management interventions.

Cerebral cortex activation mapping upon electrical muscle stimulation by 32-channel time-domain functional near-infrared spectroscopy.

The application of different EMS current thresholds on muscle activates not only the muscle but also peripheral sensory axons that send proprioceptive and pain signals to the cerebral cortex. A 32-channel time-domain fNIRS instrument was employed to map regional cortical activities under varied EMS current intensities applied on the right wrist extensor muscle. Eight healthy volunteers underwent four EMS at different current thresholds based on their individual maximal tolerated intensity (MTI), i.e., 10 % < 50 % < 100 % < over 100 % MTI. Time courses of the absolute oxygenated and deoxygenated hemoglobin concentrations primarily over the bilateral sensorimotor cortical (SMC) regions were extrapolated, and cortical activation maps were determined by general linear model using the NIRS-SPM software. The stimulation-induced wrist extension paradigm significantly increased activation of the contralateral SMC region according to the EMS intensities, while the ipsilateral SMC region showed no significant changes. This could be due in part to a nociceptive response to the higher EMS current intensities and result also from increased sensorimotor integration in these cortical regions.

Robustness of tissue oxygenation estimates by continuous wave space-resolved near infrared spectroscopy.

Significance: Continuous wave near infrared spectroscopy (CW-NIRS) is widely exploited in clinics to estimate skeletal muscles and brain cortex oxygenation. Spatially resolved spectroscopy (SRS) is generally implemented in commercial devices. However, SRS suffers from two main limitations: the a priori assumption on the spectral dependence of the reduced scattering coefficient [µs'(λ)] and the modeling of tissue as homogeneous. Aim: We studied the accuracy and robustness of SRS NIRS. We investigated the errors in retrieving hemodynamic parameters, in particular tissue oxygen saturation (StO2), when µs'(λ) was varied from expected values, and when layered tissue was considered. Approach: We simulated hemodynamic variations mimicking real-life scenarios for skeletal muscles. Simulations were performed by exploiting the analytical solutions of the photon diffusion equation in different geometries: (1) semi-infinite homogeneous medium and constant µs'(λ); (2) semi-infinite homogeneous medium and linear changes in µs'(λ); (3) two-layered media with a superficial thickness s1=5, 7.5, 10 mm and constant µs'(λ). All simulated data were obtained at source-detector distances ρ=35, 40, 45 mm, and analyzed with the SRS approach to derive hemodynamic parameters (concentration of oxygenated and deoxygenated hemoglobin, total hemoglobin concentration, and tissue oxygen saturation, StO2) and their relative error. Results: Variations in µs'(λ) affect the estimated StO2 (up to ±10%), especially if changes are different at the two wavelengths. However, the main limitation of the SRS method is the presence of a superficial layer: errors strongly larger than 20% were retrieved for the estimated StO2 when the superficial thickness exceeds 5 mm. Conclusions: These results highlight the need for more sophisticated strategies (e.g., the use of multiple short and long distances) to reduce the influence of superficial tissues in retrieving hemodynamic parameters and warn the SRS users to be aware of the intrinsic limitation of this approach, particularly when exploited in the clinical environment.

Motor cortex hemodynamic response to goal-oriented and non-goal-oriented tasks in healthy subjects.

Background: Motor disorders are one of the world's major scourges, and neuromotor rehabilitation is paramount for prevention and monitoring plans. In this scenario, exercises and motor tasks to be performed by patients are crucial to follow and assess treatments' progression and efficacy. Nowadays, in clinical environments, quantitative assessment of motor cortex activities during task execution is rare, due to the bulkiness of instrumentation and the need for immobility during measurements [e.g., functional magnetic resonance imaging (MRI)]. Functional near-infrared spectroscopy (fNIRS) can contribute to a better understanding of how neuromotor processes work by measuring motor cortex activity non-invasively in freely moving subjects. Aim: Exploit fNIRS to measure functional activation of the motor cortex area during arm-raising actions. Design: All subjects performed three different upper limbs motor tasks: arm raising (non-goal-oriented), arm raising and grasping (goal oriented), and assisted arm raising (passive task). Each task was repeated ten times. The block design for each task was divided into 5 seconds of baseline, 5 seconds of activity, and 15 seconds of recovery. Population: Sixteen healthy subjects (11 males and 5 females) with an average (+/- standard deviation) of 37.9 (+/- 13.0) years old. Methods: Cerebral hemodynamic responses have been recorded in two locations, motor cortex (activation area) and prefrontal cortex (control location) exploiting commercial time-domain fNIRS devices. Haemodynamic signals were analyzed, separating the brain cortex hemodynamic response from extracerebral hemodynamic variations. Results: The hemodynamic response was recorded in the cortical motor area for goal-oriented and not-goaloriented tasks, while no response was noticed in the control location (prefrontal cortex position). Conclusions: This study provides a basis for canonical upper limb motor cortex activations that can be potentially compared to pathological cerebral responses in patients. It also highlights the potential use of TD-fNIRS to study goal-oriented versus non-goaloriented motor tasks. Impact: the findings of this study may have implications for clinical rehabilitation by providing a better understanding of the neural mechanisms underlying goal-oriented versus non-goal-oriented motor tasks. This may lead to more effective rehabilitation strategies for individuals with motor disorders and a more effective diagnosis of motor dysfunction supported by objective and quantitative neurophysiological readings.

Quality Characteristics, Sensory Profiles and Ethylene Production of Stored 'Abate Fetel' Pears Sorted at Harvest by Time-Resolved Reflectance Spectroscopy.

Time-resolved reflectance spectroscopy (TRS), a nondestructive technique, can help the industry to provide high-quality fruit to encourage pear consumption. The absorption coefficient measured by TRS at 670 nm (µa670) represents a maturity index for pear fruit, with less mature pears high µa670 and more mature low µa670. The aim of this work was to study the quality characteristics, the sensory profiles and the ethylene production of 'Abate Fetel' pears sorted at harvest in different TRS maturity classes and stored in different atmospheres. At harvest, 540 pears were measured by TRS for µa670, ranked by µa670 in three maturity classes (less-LeM, medium-MeM and more-MoM mature) and randomized in nine samples according to 1-MCP treatment (treated, control), storage time (4-6 months) and atmosphere (air-NA; CA: 8-12 kPa O2, 1 kPa CO2). Fruits were examined at harvest and after 7 days of poststorage shelf life for skin color, firmness, soluble solids, acidity and ethylene production and were submitted to sensory analysis. At harvest and after storage, MoM pears were less green and showed a higher SSC content than LeM ones. After storage, MoM pears produced less ethylene and were perceived to be firmer (especially in 1-MCP-treated pears), more astringent and less juicy (when stored for 6 months) than LeM ones.

Fast time-domain diffuse correlation spectroscopy with superconducting nanowire single-photon detector: system validation and in vivo results.

Time-domain diffuse correlation spectroscopy (TD-DCS) has been introduced as an advancement of the "classical" continuous wave DCS (CW-DCS) allowing one to not only to measure depth-resolved blood flow index (BFI) but also to extract optical properties of the measured medium without using any additional diffuse optics technique. However, this method is a photon-starved technique, specially when considering only the late photons that are of primary interest which has limited its in vivo application. In this work, we present a TD-DCS system based on a superconducting nanowire single-photon detector (SNSPD) with a high quantum efficiency, a narrow timing response, and a negligibly low dark count noise. We compared it to the typically used single-photon avalanche diode (SPAD) detector. In addition, this system allowed us to conduct fast in vivo measurements and obtain gated pulsatile BFI on the adult human forehead.

Cerebrovascular reactivity to carbon dioxide tension in newborns: data from combined time-resolved near-infrared spectroscopy and diffuse correlation spectroscopy.

Significance: Critically ill newborns are at risk of brain damage from cerebrovascular disturbances. A cerebral hemodynamic monitoring system would have the potential role to guide targeted intervention. Aim: To obtain, in a population of newborn infants, simultaneous near-infrared spectroscopy (NIRS)-based estimates of cerebral tissue oxygen saturation (StO2) and blood flow during variations of carbon dioxide tension (pCO2) levels within physiologic values up to moderate permissive hypercapnia, and to examine if the derived estimate of metabolic rate of oxygen would stay constant, during the same variations. Approach: We enrolled clinically stable mechanically ventilated newborns at postnatal age >24 h without brain abnormalities at ultrasound. StO2 and blood flow index were measured using a non-invasive device (BabyLux), which combine time-resolved NIRS and diffuse-correlation spectroscopy. The effect of changes in transcutaneous pCO2 on StO2, cerebral blood flow (CBF), and cerebral metabolic rate of oxygen index (tCMRO2i) were estimated. Results: Ten babies were enrolled and three were excluded. Median GA at enrollment was 39 weeks and median weight 2720 g. StO2 increased 0.58% (95% CI 0.55; 0.61, p<0.001), CBF 2% (1.9; 2.3, p<0.001), and tCMRO2 0.3% (0.05; 0.46, p=0.017) per mmHg increase in pCO2. Conclusions: BabyLux device detected pCO2-induced changes in cerebral StO2 and CBF, as expected. The small statistically significant positive relationship between pCO2 and tCMRO2i variation is not considered clinically relevant and we are inclined to consider it as an artifact.