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Correlative all-optical quantification of mass density and mechanics of sub-cellular compartments with fluorescence specificity

Schlü�ler, R and Kim, K and Nötzel, M and Taubenberger, A and Abuhattum, S and Beck, T and Müller, P and Maharana, S and Cojoc, G and Girardo, S and Hermann, A and Alberti, S and Guck, J (2022) Correlative all-optical quantification of mass density and mechanics of sub-cellular compartments with fluorescence specificity. In: eLife, 11 .

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Official URL: https://doi.org/10.7554/eLife.68490


Quantitative measurements of physical parameters become increasingly important for understanding biological processes. Brillouin microscopy (BM) has recently emerged as one technique providing the 3D distribution of viscoelastic properties inside biological samples — so far relying on the implicit assumption that refractive index (RI) and density can be neglected. Here, we present a novel method (FOB microscopy) combining BM with optical diffraction tomography and epi-fluorescence imaging for explicitly measuring the Brillouin shift, RI and absolute density with specificity to fluorescently labeled structures. We show that neglecting the RI and density might lead to erroneous conclusions. Investigating the nucleoplasm of wild-type HeLa cells, we find that it has lower density but higher longitudinal modulus than the cytoplasm. Thus, the longitudinal modulus is not merely sensitive to the water content of the sample — a postulate vividly discussed in the field. We demonstrate the further utility of FOB on various biological systems including adipocytes and intracellular membraneless compartments. FOB microscopy can provide unexpected scientific discoveries and shed quantitative light on processes such as phase separation and transition inside living cells.

Item Type: Journal Article
Publication: eLife
Publisher: eLife Sciences Publications Ltd
Additional Information: The copyright for this article belongs to the Authors.
Keywords: adipocyte; article; biology; controlled study; cytoplasm; diffraction; fluorescence imaging; human cell; human tissue; intracellular space; microscopy; phase separation; refraction index; tomography; water content; wild type; cell nucleus; cells; cytology; fluorescence; HeLa cell line; human; optical tomography; procedures; refractometry; ultrastructure, Cell Nucleus; Cells; Fluorescence; HeLa Cells; Humans; Intracellular Space; Microscopy; Refractometry; Tomography, Optical
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 14 Jul 2022 04:41
Last Modified: 14 Jul 2022 04:41
URI: https://eprints.iisc.ac.in/id/eprint/74341

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