Label-free hyperspectral dark-field microscopy towards quantitative scatter imaging

Published

March 27, 2017

Author(s)

Philip Cheney, David McClatchy III, Stephen Kanick, Paul Lemaillet, David W. Allen, Daniel Samarov, Brian Pogue, Jeeseong C. Hwang

Abstract

A hyperspectral dark-field microscopy technique has been developed for imaging spatially distributed diffuse reflectance spectra from light-scattering samples. In this report, quantitative scatter spectroscopy was demonstrated with a uniform scattering phantom, a solution of polystyrene microspheres. A Monte Carlo-based inverse model was used to calculate the reduced scattering coefficients of the samples of different microsphere concentrations from wavelength-dependent backscattered signal measured by the dark-field microscopy. The result was compared with the measurement result by a NIST's double integrating sphere system for validation. Ongoing efforts involve quantitative mapping of scattering and absorption coefficients in samples with spatially heterogeneous optical properties.

Citation

Proceedings of SPIE

Volume

10056

Pub Type

Journals

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DOI Link

Keywords

Tissue-simulating phantoms, hyperspectral dark-field microscopy, scatter imaging, integrating sphere, Monte Carlo simulation, scattering coefficient

Citation

Cheney, P. , McClatchy III, D. , Kanick, S. , Lemaillet, P. , Allen, D. , Samarov, D. , Pogue, B. and Hwang, J. (2017), Label-free hyperspectral dark-field microscopy towards quantitative scatter imaging, Proceedings of SPIE, [online], https://doi.org/10.1117/12.2263336 (Accessed October 8, 2025)