To approach wide-field optical properties quantification in real heterogeneous biological tissue, we developed a Dual-Step setup that couples a punctual diffuse reflectance spectroscopy (DRS) technique with multispectral imaging (MSI). The setup achieves wide-field optical properties assessment through an initial estimation of scattering with DRS, which is used to estimate absorption with MSI. The absolute quantification of optical properties is based on the ACA-Pro algorithm that has been adapted both for DRS and for MSI. This paper validates the Dual-Step system not only on homogeneous Intralipid phantoms but also on a heterogeneous gelatine phantom with different scattering and absorbing properties.
We present the Dual-Step system and method we developed to achieve 2D quantitative maps of optical properties. It is non-contact, quantitative for both absorption and scattering, large field, and spectrally resolved. The present study shows the results obtained on rats and figures the interest of the approach to address complex in-vivo samples.
We have developed an adaptive calibration algorithm and protocol (ACA-Pro) that corrects from the instrumental response of various spatially resolved diffuse reflectance spectroscopy (DRSsr) systems to enable the quantification of absorption and scattering properties based on a Monte Carlo-based look-up-table approach. The protocol involves the use of a calibration reference base built with measurements of a range of different diffusive intralipid phantoms. Moreover, an advanced strategy was established to take into account the experimental variations with an additional measurement of a common solid material, allowing the use of a single calibration reference base for all experiments. The ACA-Pro is validated in contact and noncontact probe-based DRSsr systems. Furthermore, the first results of a setup replacing the probe with a CCD detector are shown to confirm the robustness of the approach.
KEYWORDS: Optical properties, Algorithm development, Monte Carlo methods, Imaging systems, Signal detection, Optical transfer functions, Spectroscopy, Absorption, Scattering, Electroluminescent displays
We validate a non-contact Diffuse Reflectance Spectroscopy (DRS) system as a first stage to approach quantitative multi-spectral imaging technique. The non-contact DRS system with separated illumination and detection paths was developed with different progressive set-ups which were all compared to a well-founded contact DRS system. While quantitation of the absorption coefficient is well achieved with the existing method, the calculation of the scattering coefficient is deteriorated by the non-contact architecture measurements. We have therefore developed an adaptive reference-based algorithm to compensate for this effect.
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