3D refractive index tomography with quantitative oblique back-illumination microscopy

Patrick B. Ledwig; Francisco E. Robles

doi:10.1117/12.2544328

17 February 2020 3D refractive index tomography with quantitative oblique back-illumination microscopy

Proceedings Volume 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII; 1124503 (2020) https://doi.org/10.1117/12.2544328
Event: SPIE BiOS, 2020, San Francisco, California, United States

Abstract

We have recently developed quantitative oblique back-illumination microscopy (qOBM), which enables full-field quantitative phase imaging (QPI) of objects embedded in a thick scattering medium. This epi-mode technique makes use of multiple scattering as a source of transmissive illumination from within, allowing for rich structural detail based on optical path delay. We now produce quantitative 3D renderings of index of refraction with sub- cellular detail by computing a 3D transfer function of the entire optical system, including the multiply scattered illumination, and deconvolving it from a vertical stack of phase gradient contrast images. This approach allows truly non-invasive, label-free, tomographic quantitative reconstructions if index of refraction in thick scattering samples including thick tissue samples, at low cost and with simple operation, bringing QPI’s unparalleled access to sub-cellular structural detail to previously unavailable domains of investigation.

Conference Presentation

Citation Download Citation

Patrick B. Ledwig and Francisco E. Robles "3D refractive index tomography with quantitative oblique back-illumination microscopy", Proc. SPIE 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII, 1124503 (17 February 2020); https://doi.org/10.1117/12.2544328

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