Optical density based quantification of total haemoglobin concentrations with spectroscopic optical coherence tomography.

Spectroscopic optical coherence tomography (sOCT) has emerged as a new possibility for non-invasive quantification of total haemoglobin concentrations [tHb]. Recently, we demonstrated that [tHb] measured in ex-vivo human whole-blood with a conventional sOCT system achieves a precision of 9.10 g/dL with a bias of 1.50 g/dL. This precision improved by acquiring data with a combination of focus tracking and zero-delay acquisition (FZA) that compensated for experimental limitations, increasing to 3.80 g/dL with a bias of 1.50 g/dL. Nevertheless, sOCT precision should improve at least to [Formula: see text] g/dL to be clinically relevant. Therefore, sOCT-based [tHb] determinations require the development of new analysis methods that reduce the variability of [tHb] estimations. In this work, we aim to increase sOCT precision by retrieving the [tHb] content from a numerical optimisation of the optical density (OD), while considering the blood absorption flattening effect. The OD-based approach simplifies previous two-step Lambert-Beer fitting approaches to a single step, thereby reducing errors during the fitting procedure. We validated our model with ex-vivo [tHb] measurements on flowing whole-blood samples in the clinical range (7-23 g/dL). Our results show that, with the new model, conventional sOCT can determine [tHb] with a precision of 3.09 g/dL and a bias of 0.86 g/dL compared to a commercial blood analyser. We present further precision improvement by combining the OD methodology with FZA, leading to a precision of 2.08 g/dL with a bias of 0.46 g/dL.

Microfluidic organ-on-a-chip model of the outer blood-retinal barrier with clinically relevant read-outs for tissue permeability and vascular structure.

The outer blood-retinal barrier (oBRB) tightly controls the transport processes between the neural tissue of the retina and the underlying blood vessel network. The barrier is formed by the retinal pigment epithelium (RPE), its basal membrane and the underlying choroidal capillary bed. Realistic three-dimensional cell culture based models of the oBRB are needed to study mechanisms and potential treatments of visual disorders such as age-related macular degeneration that result from dysfunction of the barrier tissue. Ideally, such models should also include clinically relevant read-outs to enable translation of experimental findings in the context of pathophysiology. Here, we report a microfluidic organ-on-a-chip model of the oBRB that contains a monolayer of human immortalized RPE and a microvessel of human endothelial cells, separated by a semi-permeable membrane. Confluent monolayers of both cell types were confirmed by fluorescence microscopy. The three-dimensional vascular structures within the chip were imaged by optical coherence tomography: a medical imaging technique, which is routinely applied in ophthalmology. Differences in diameters and vessel density could be readily detected. Upon inducing oxidative stress by treating with hydrogen peroxide (H2O2), a dose dependent increase in barrier permeability was observed by using a dynamic assay for fluorescence tracing, analogous to the clinically used fluorescence angiography. This organ-on-a-chip of the oBRB will allow future studies of complex disease mechanisms and treatments for visual disorders using clinically relevant endpoints in vitro.

Dependency of the optical scattering properties of human milk on casein content and common sample preparation methods.

SIGNIFICANCE: Quantifying human milk composition is important for daily nutritional management in neonatal intensive cares worldwide. Photonic solutions based on visible light can potentially aid in this analysis, as energy content of human milk depends largely on fat content, and the optical scattering properties of human milk predominantly depend on the size and concentration of fat globules. However, it is expected that human milk scattering changes upon homogenization, routinely done before analysis, which may affect fat globule size. AIM: The first aim of this study was to investigate how the most common homogenization methods (gently inverting by hand, vortexing, and sonication) affect the optical properties of human milk. The second aim was to estimate the scattering contribution of casein micelles, the second most dominant scatterers in human milk. APPROACH: We combined diffuse reflectance spectroscopy with spectroscopic optical coherence tomography to measure the scattering coefficient µs, reduced scattering coefficient µs', and anisotropy g between 450 and 600 nm. RESULTS: Sonication induced the strongest changes in µs, µs', and g compared to the gently inverted samples (203%, 202%, and 7%, respectively, at 550 nm), but also vortexing changed µs' with 20%. Although casein micelles only showed a modest contribution to µs and g at 550 nm (7% and 1%, respectively), their contribution to µs' was 29%. CONCLUSIONS: The scattering properties of human milk strongly depend on the homogenization method that is employed, and gentle inversion should be the preferred method. The contribution of casein micelles was relatively small for µs and g but considerably larger for µs'.

Quantification of total haemoglobin concentrations in human whole blood by spectroscopic visible-light optical coherence tomography.

The non-invasive quantification of total haemoglobin concentrations [tHb] is highly desired for the assessment of haematologic disorders in vulnerable patient groups, but invasive blood sampling is still the gold standard in current clinical practice. This work demonstrates the potential of visible-light spectroscopic optical coherence tomography (sOCT) for quantifying the [tHb] in human whole blood. To accurately quantify the [tHb] from the substantial optical attenuation by blood in the visible wavelength range, we used a combination of zero-delay acquisition and focus tracking that ensures optimal system sensitivity at any depth inside the sample. Subsequently, we developed an analysis model to adequately correct for the high scattering contribution by red blood cells to the sOCT signal. We validate our method and compare it to conventional sOCT (without focus tracking and zero-delay acquisition) through ex-vivo measurements on flowing human whole blood, with [tHb] values in the clinical range of 7-23 g/dL. For our method with optimized sensitivity, the measured and expected values correlate well (Pearson correlation coefficient = 0.89, p < 0.01), with a precision of 3.8 g/dL. This is a considerable improvement compared to conventional sOCT (Pearson correlation coefficient = 0.59, p = 0.16; precision of 9.1 g/dL).

Optical properties of human milk.

With human milk being the most important source of infant nutrition, the protection and support of breastfeeding are essential from a global health perspective. Nevertheless, relatively few objective methods are available to investigate human milk composition and lactation physiology when a mother experiences breastfeeding problems. We argue that optics and photonics offer promising opportunities for this purpose. Any research activity within this new application field starts with a thorough understanding on how light interacts with human milk. Therefore, the aim of this study was to investigate the full set of optical properties for human milk and the biological variability therein. Using a novel approach that combines spatially resolved diffuse reflectance spectroscopy (SR-DRS) and spectroscopic optical coherence tomography (sOCT) between 450 and 650 nm, we quantified the absorption coefficient µa , scattering coefficient µs , reduced scattering coefficient µs', anisotropy g and backscattering coefficient µb,NA of mature human milk from 14 participants released at different stages during a breastfeed (foremilk, bulk milk and hindmilk). Significant correlations were found between µa , µs , µs' and µb,NA and the biochemically determined fat concentration per sample (Rs = 0.38, Rs = 0.77, Rs = 0.80, Rs = 0.44 respectively). We explained the observed variations in the optical properties of human milk using Mie theory and the biological variability in both the concentration and size distribution of milk fat globules. In conclusion, we have provided a full set of optical properties for human milk, which can hopefully serve as a starting point for future biophotonic studies on human milk and the milk containing lactating breast.

Spatially confined quantification of bilirubin concentrations by spectroscopic visible-light optical coherence tomography.

Spatially confined measurements of bilirubin in tissue can be of great value for noninvasive bilirubin estimations during neonatal jaundice, as well as our understanding of the physiology behind bilirubin extravasation. This work shows the potential of spectroscopic visible-light optical coherence tomography (sOCT) for this purpose. At the bilirubin absorption peak around 460 nm, sOCT suffers from a strong signal decay with depth, which we overcome by optimizing our system sensitivity through a combination of zero-delay acquisition and focus tracking. In a phantom study, we demonstrate the quantification of bilirubin concentrations between 0 and 650 µM with only a 10% difference to the expected value, thereby covering the entire clinical pathophysiological range.

Measuring absorption coefficient of scattering liquids using a tube inside an integrating sphere.

A method for measuring the absorption coefficient µa of absorbing and scattering liquid samples is presented. The sample is injected into a small transparent tube mounted through an integrating sphere. Two models for determining the absorption coefficient using the relative optical output signal are described and validated using aqueous ink absorbers of 0.5 vol.% (0.3 mm-1<µa<1.55 mm-1) and 1.0 vol.% (1.0 mm-1<µa<4.0 mm-1) concentrations with 1 vol.% (µs'≈1.4 mm-1) and 10 vol.% (µs'≈14 mm-1) Intralipid dilutions. The low concentrations give µa and µs values, which are comparable with those of biological tissues. One model assumes a uniform light distribution within the sample, which is valid for low absorption. Another model considers light attenuation that obeys Lambert-Beer's law, which may be used for relatively high absorption. Measurements with low and high scattering samples are done for the wavelength range of 400-900 nm. Measured spectra of purely absorbing samples are within 15% agreement with measurements using standard transmission spectrophotometry. For 0.5 vol.% ink absorbers and at wavelengths below 700 nm, measured µa values are higher for samples with low scattering and lower for those with high scattering. At wavelengths above 700 nm, measured µa values do not vary significantly with amount of scattering. For 1.0 vol.% ink absorbers, measured spectra do not change with low scattering. These results indicate that the method can be used for measuring absorption spectra of scattering liquid samples with optical properties similar to biological absorbers, particularly at wavelengths above 700 nm, which is difficult to accomplish with standard transmission spectrophotometry.