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



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


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.
Proceedings of SPIE


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


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], (Accessed June 25, 2024)


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Created March 26, 2017, Updated October 12, 2021