Refractive index-matching a cell’s immersion medium isolates organelle scattering and increases QPI contrast

Kaitlin J. Dunn; Christopher L. Weil; Andrew J. Berger

doi:10.1117/12.2631742

2 March 2022 Refractive index-matching a cell’s immersion medium isolates organelle scattering and increases QPI contrast

Kaitlin J. Dunn, Christopher L. Weil, Andrew J. Berger

Author Affiliations +

Proceedings Volume PC11970, Quantitative Phase Imaging VIII; PC1197002 (2022) https://doi.org/10.1117/12.2631742
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

The substantial refractive index contrast between a cell’s cytoplasm and traditional immersion media scatters enough light to obscure the angular scattering from cellular organelles and to reduce contrast in quantitative phase images (QPI). To reduce the whole-cell scattering, we mix cell media with a biocompatible, high-refractive-index liquid to closely match the cytoplasm’s refractive index. We demonstrate with live single-cell images that this enables isolation of organelles’ angular scattering, which will improve angular scattering-based estimates of organelle size. We also explain why index-matching enhances phase contrast within a cell’s QPI and propose a digital method for obtaining this result.

Conference Presentation

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Kaitlin J. Dunn, Christopher L. Weil, and Andrew J. Berger "Refractive index-matching a cell’s immersion medium isolates organelle scattering and increases QPI contrast", Proc. SPIE PC11970, Quantitative Phase Imaging VIII, PC1197002 (2 March 2022); https://doi.org/10.1117/12.2631742

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