Scattered light contrast microscopy for reflectance imaging of thick tissue (Conference Presentation)

Jeremy D. Rogers

doi:10.1117/12.2513282

4 March 2019 Scattered light contrast microscopy for reflectance imaging of thick tissue (Conference Presentation)

Jeremy D. Rogers

Proceedings Volume 10890, Label-free Biomedical Imaging and Sensing (LBIS) 2019; 1089019 (2019) https://doi.org/10.1117/12.2513282
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

Label-free methods provide image contrast without the need to introduce exogenous material that may alter the sample microstructure, perturb the microenvironment, or limit clinical translation. In addition to being label-free, it is increasingly important to consider modalities that work in reflection mode geometry to allow imaging intact tissue (in vivo or ex vivo) or 3D cell cultures too thick to image in transmission. However, reflection geometry precludes many useful contrast modalities, including transmitted phase and darkfield. Optical scattering in tissue limits contrast and depth of imaging, but scattering can also be exploited to quantify tissue structure and provide diagnostic markers. Here, we present another application of multiple scattering in tissue in which collection of scattered light is used to reconstruct images with contrast comparable to transmission imaging modalities. A Scattered Light Contrast (SLC) microscope collects a descanned image of the scattered light surrounding the confocal illumination point. The illumination point is raster scanned and the scattered light distribution is collected for each image pixel, providing 4D data. By analyzing the scattered light, a range of image contrast modalities can be reconstructed including reflectance confocal microscopy, transmitted phase contrast, and dark field microscopy. Computational Monte Carlo simulations relate the SLC signal to the phase gradient in the image plane and can be used to reconstruct the phase gradient of transmitted light in the focal plane. This modality enables label-free imaging of cells and structures in the retina that will advance diagnosis and monitoring therapies at the cellular level.

Conference Presentation

Citation Download Citation

Jeremy D. Rogers "Scattered light contrast microscopy for reflectance imaging of thick tissue (Conference Presentation)", Proc. SPIE 10890, Label-free Biomedical Imaging and Sensing (LBIS) 2019, 1089019 (4 March 2019); https://doi.org/10.1117/12.2513282

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KEYWORDS

Light scattering

Tissue optics

Microscopy

Reflectivity

Image transmission

Confocal microscopy

In vivo imaging

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