Two-layered blood-lipid phantom and method to determine absorption and oxygenation employing changes in moments of DTOFs.

Near-infrared spectroscopy (NIRS) is an established technique for measuring tissue oxygen saturation (StO2), which is of high clinical value. For tissues that have layered structures, it is challenging but clinically relevant to obtain StO2 of the different layers, e.g. brain and scalp. For this aim, we present a new method of data analysis for time-domain NIRS (TD-NIRS) and a new two-layered blood-lipid phantom. The new analysis method enables accurate determination of even large changes of the absorption coefficient (Δµa) in multiple layers. By adding Δµa to the baseline µa, this method provides absolute µa and hence StO2 in multiple layers. The method utilizes (i) changes in statistical moments of the distributions of times of flight of photons (DTOFs), (ii) an analytical solution of the diffusion equation for an N-layered medium, (iii) and the Levenberg-Marquardt algorithm (LMA) to determine Δµa in multiple layers from the changes in moments. The method is suitable for NIRS tissue oximetry (relying on µa) as well as functional NIRS (fNIRS) applications (relying on Δµa). Experiments were conducted on a new phantom, which enabled us to simulate dynamic StO2 changes in two layers for the first time. Two separate compartments, which mimic superficial and deep layers, hold blood-lipid mixtures that can be deoxygenated (using yeast) and oxygenated (by bubbling oxygen) independently. Simultaneous NIRS measurements can be performed on the two-layered medium (variable superficial layer thickness, L), the deep (homogeneous), and/or the superficial (homogeneous). In two experiments involving ink, we increased the nominal µa in one of two compartments from 0.05 to 0.25 cm-1, L set to 14.5 mm. In three experiments involving blood (L set to 12, 15, or 17 mm), we used a protocol consisting of six deoxygenation cycles. A state-of-the-art multi-wavelength TD-NIRS system measured simultaneously on the two-layered medium, as well as on the deep compartment for a reference. The new method accurately determined µa (and hence StO2) in both compartments. The method is a significant progress in overcoming the contamination from the superficial layer, which is beneficial for NIRS and fNIRS applications, and may improve the determination of StO2 in the brain from measurements on the head. The advanced phantom may assist in the ongoing effort towards more realistic standardized performance tests in NIRS tissue oximetry. Data and MATLAB codes used in this study were made publicly available.

Depth-selective method for time-domain diffuse reflectance measurements: validation study of the dual subtraction technique.

Research on the spatial distribution of sensitivity of time-domain near infrared diffuse reflectance measurement is reported in this paper. The main objective of the investigation is to validate theoretically calculated sensitivity profiles for a measurement geometry with two detectors and two sources in which sensitivity profiles of statistical moments of distributions of time of flight of photons (DTOFs) are spatially restricted to a region underneath the detectors. For this dual subtraction method, smaller sensitivities to changes appearing in the superficial layer of the medium were observed compared to the single distance and single subtraction methods. Experimental validation of this approach is based on evaluation of changes in the statistical moments of DTOFs measured on a liquid phantom with local absorption perturbations. The spatial distributions of sensitivities, depth-related sensitivity and depth selectivities were obtained from the dual subtraction method and compared with those from single distance and single subtraction approaches. Also, the contrast to noise ratio (CNR) was calculated for the dual subtraction technique and combined with depth selectivity in order to assess the overall performance (product of CNR and depth selectivity) of the method. Spatial sensitivity profiles from phantom experiments are in a good agreement with the results of theoretical studies and feature more locally restricted sensitivity volume with the point of maximal sensitivity located deeper. The highest value of overall performance was obtained experimentally for the second statistical moment in the dual subtraction method (∼10.8) surpassing that of the single distance method (∼8.7). This confirms the advantage of dual subtraction measurement geometries in the suppression of optical signals originated in the superficial layer of the medium.

Multi-laboratory performance assessment of diffuse optics instruments: the BitMap exercise.

SIGNIFICANCE: Multi-laboratory initiatives are essential in performance assessment and standardization-crucial for bringing biophotonics to mature clinical use-to establish protocols and develop reference tissue phantoms that all will allow universal instrument comparison. AIM: The largest multi-laboratory comparison of performance assessment in near-infrared diffuse optics is presented, involving 28 instruments and 12 institutions on a total of eight experiments based on three consolidated protocols (BIP, MEDPHOT, and NEUROPT) as implemented on three kits of tissue phantoms. A total of 20 synthetic indicators were extracted from the dataset, some of them defined here anew. APPROACH: The exercise stems from the Innovative Training Network BitMap funded by the European Commission and expanded to include other European laboratories. A large variety of diffuse optics instruments were considered, based on different approaches (time domain/frequency domain/continuous wave), at various stages of maturity and designed for different applications (e.g., oximetry, spectroscopy, and imaging). RESULTS: This study highlights a substantial difference in hardware performances (e.g., nine decades in responsivity, four decades in dark count rate, and one decade in temporal resolution). Agreement in the estimates of homogeneous optical properties was within 12% of the median value for half of the systems, with a temporal stability of <5 % over 1 h, and day-to-day reproducibility of <3 % . Other tests encompassed linearity, crosstalk, uncertainty, and detection of optical inhomogeneities. CONCLUSIONS: This extensive multi-laboratory exercise provides a detailed assessment of near-infrared Diffuse optical instruments and can be used for reference grading. The dataset-available soon in an open data repository-can be evaluated in multiple ways, for instance, to compare different analysis tools or study the impact of hardware implementations.

Time-domain NIRS system based on supercontinuum light source and multi-wavelength detection: validation for tissue oxygenation studies.

We present and validate a multi-wavelength time-domain near-infrared spectroscopy (TD-NIRS) system that avoids switching wavelengths and instead exploits the full capability of a supercontinuum light source by emitting and acquiring signals for the whole chosen range of wavelengths. The system was designed for muscle and brain oxygenation monitoring in a clinical environment. A pulsed supercontinuum laser emits broadband light and each of two detection modules acquires the distributions of times of flight of photons (DTOFs) for 16 spectral channels (used width 12.5 nm / channel), providing a total of 32 DTOFs at up to 3 Hz. Two emitting fibers and two detection fiber bundles allow simultaneous measurements at two positions on the tissue or at two source-detector separations. Three established protocols (BIP, MEDPHOT, and nEUROPt) were used to quantitatively assess the system's performance, including linearity, coupling, accuracy, and depth sensitivity. Measurements were performed on 32 homogeneous phantoms and two inhomogeneous phantoms (solid and liquid). Furthermore, measurements on two blood-lipid phantoms with a varied amount of blood and Intralipid provide the strongest validation for accurate tissue oximetry. The retrieved hemoglobin concentrations and oxygen saturation match well with the reference values that were obtained using a commercially available NIRS system (OxiplexTS) and a blood gas analyzer (ABL90 FLEX), except a discrepancy occurs for the lowest amount of Intralipid. In-vivo measurements on the forearm of three healthy volunteers during arterial (250 mmHg) and venous (60 mmHg) cuff occlusions provide an example of tissue monitoring during the expected hemodynamic changes that follow previously well-described physiologies. All results, including quantitative parameters, can be compared to other systems that report similar tests. Overall, the presented TD-NIRS system has an exemplary performance evaluated with state-of-the-art performance assessment methods.

Space-enhanced time-domain diffuse optics for determination of tissue optical properties in two-layered structures.

A novel methodology for solving the inverse problem of diffuse optics for two-layered structures is proposed to retrieve the absolute quantities of optical absorption and reduced scattering coefficients of the layers simultaneously. A liquid phantom with various optical absorption properties in the deep layer is prepared and experimentally investigated using the space-enhanced time-domain method. Monte-Carlo simulations are applied to analyze the different measurements in time domain, space domain, and by the new methodology. The deviations of retrieved values from nominal values of both layers' optical properties are simultaneously reduced to a very low extent compared to the single-domain methods. The reliability and uncertainty of the retrieval performance are also considerably improved by the new methodology. It is observed in time-domain analyses that for the deep layer the retrieval of absorption coefficient is almost not affected by the scattering properties and this kind of "deep scattering neutrality" is investigated and overcome as well.

Performance of measurands in time-domain optical brain imaging: depth selectivity versus contrast-to-noise ratio.

Time-domain optical brain imaging techniques introduce a number of different measurands for analyzing absorption changes located deep in the tissue, complicated by superficial absorption changes and noise. We implement a method that allows analysis, quantitative comparison and performance ranking of measurands under various conditions - including different values of reduced scattering coefficient, thickness of the superficial layer, and source-detector separation. Liquid phantom measurements and Monte Carlo simulations were carried out in two-layered geometry to acquire distributions of times of flight of photons and to calculate the total photon count, mean time of flight, variance, photon counts in time windows and ratios of photon counts in different time windows. Quantitative comparison of performance was based on objective metrics: relative contrast, contrast-to-noise ratio (CNR) and depth selectivity. Moreover, the product of CNR and depth selectivity was used to rank the overall performance and to determine the optimal source-detector separation for each measurand. Variance ranks the highest under all considered conditions.

Near-Infrared Spectroscopy Measured Cerebral Blood Flow from Spontaneous Oxygenation Changes in Neonatal Brain Injury.

Neonates with hypoxic-ischaemic (HI) brain injury were monitored using a broadband near-infrared spectroscopy (NIRS) system in the neonatal intensive care unit. The aim of this work is to use the NIRS cerebral oxygenation data (HbD = oxygenated-haemoglobin - deoxygenated-haemoglobin) combined with arterial saturation (SaO2) from pulse oximetry to calculate cerebral blood flow (CBF) based on the oxygen swing method, during spontaneous desaturation episodes. The method is based on Fick's principle and uses HbD as a tracer; when a sudden change in SaO2 occurs, the change in HbD represents a change in tracer concentration, and thus it is possible to estimate CBF. CBF was successfully calculated with broadband NIRS in 11 HIE infants (3 with severe injury) for 70 oxygenation events on the day of birth. The average CBF was 18.0 ± 12.7 ml 100 g-1 min-1 with a range of 4 ml 100 g-1 min-1 to 60 ml 100 g-1 min-1. For infants with severe HIE (as determined by magnetic resonance spectroscopy) CBF was significantly lower (p = 0.038, d = 1.35) than those with moderate HIE on the day of birth.

Influence of contrast-reversing frequency on the amplitude and spatial distribution of visual cortex hemodynamic responses.

Visual stimulation is one of the most commonly used paradigms for cerebral cortex function investigation. Experiments typically involve presenting to a volunteer a black-and-white checkerboard with contrast-reversing at a frequency of 4 to 16 Hz. The aim of the present study was to investigate the influence of the flickering frequency on the amplitude of changes in the concentration of oxygenated and deoxygenated hemoglobin. The hemoglobin concentrations were assessed with the use of a high resolution diffuse optical tomography method. Spatial distributions of changes in hemoglobin concentrations overlaying the visual cortex are shown for various stimuli frequencies. Moreover, the hemoglobin concentration changes obtained for different source-detector separations (from 1.5 to 5.4 cm) are presented. Our results demonstrate that the flickering frequency had a statistically significant effect on the induced oxyhemoglobin changes (p < 0,001). The amplitude of oxy hemoglobin concentration changes at a frequency of 8 Hz was higher in comparison with that measured at 4 Hz :[median(25th-75thpercentiles) 1.24 (0.94-1.71) vs. 0.92(0.73-1.28)µM, p < 0.001]; 12 Hz:[1.24 (0.94-1.71) vs. 1.04 (0.78-1.32) µM, p < 0.001]; and 16 Hz:[1.24 (0.94-1.71) vs. 1.15(0.87-1.48) µM, p < 0.001]. No significant differences were observed between the size of an area of activation for various frequencies. The demonstrated superiority of 8 Hz over other frequencies can advance understanding of visual stimulations and help guide future fNIRS protocols.

Depth-resolved assessment of changes in concentration of chromophores using time-resolved near-infrared spectroscopy: estimation of cytochrome-c-oxidase uncertainty by Monte Carlo simulations.

Time-resolved near-infrared spectroscopy (TR-NIRS) measurements can be used to recover changes in concentrations of tissue constituents ( Δ C ) by applying the moments method and the Beer-Lambert law. In this work we carried out the error propagation analysis allowing to calculate the standard deviations of uncertainty in estimation of the Δ C . Here, we show the process of choosing wavelengths for the evaluation of hemodynamic (oxy-, deoxyhemoglobin) and metabolic (cytochrome-c-oxidase (CCO)) responses within the brain tissue as measured with an in-house developed TR-NIRS multi-wavelength system, which measures at 16 consecutive wavelengths separated by 12.5 nm and placed between 650 and 950 nm. Data generated with Monte Carlo simulations on three-layered model (scalp, skull, brain) for wavelengths range from 650 to 950 nm were used to carry out the error propagation analysis for varying choices of wavelengths. For a detector with a spectrally uniform responsivity, the minimal standard deviation of the estimated changes in CCO within the brain layer, σ Δ C CCO brain = 0.40 µM, was observed for the 16 consecutive wavelengths from 725 to 912.5 nm. For realistic a detector model, i.e. the spectral responsivity characteristic is considered, the minimum, σ Δ C CCO brain = 0.47 µM, was observed at the 16 consecutive wavelengths from 688 to 875 nm. We introduce the method of applying the error propagation analysis to data as measured with spectral TR-NIRS systems to calculate uncertainty of recovery of tissue constituents concentrations.

Self-calibrating time-resolved near infrared spectroscopy.

Time-resolved near infrared spectroscopy is considered to be a gold standard technique when measuring absolute values of tissue optical properties, as it provides separable and independent information about both tissue absorption and scattering. However, time-resolved instruments require an accurate characterization by measuring the instrument response function in order to decouple the contribution of the instrument itself from the measurement. In this work, a new approach to the methodology of analysing time-resolved data is presented where the influence of instrument response function is eliminated from the data and a self-calibrating analysis is proposed. The proposed methodology requires an instrument to provide at least two wavelengths and allows spectral parameters recovery (optical properties or constituents concentrations and reduced scatter amplitude and power). Phantom and in-vivo data from two different time-resolved systems are used to validate the accuracy of the proposed self-calibrating approach, demonstrating that parameters recovery compared to the conventional curve fitting approach is within 10% and benefits from introducing a spectral constraint to the reconstruction problem. It is shown that a multi-wavelength time-resolved data can be used for parameters recovery directly without prior calibration (instrument response function measurement).