Correlative all-optical quantification of mass density and mechanics of subcellular compartments with fluorescence specificity
| Autor: | Kyoohyun Kim, Raimund Schlüßler, Martin Nötzel, Anna Taubenberger, Shada Abuhattum, Timon Beck, Paul Müller, Shovamaye Maharana, Gheorghe Cojoc, Salvatore Girardo, Andreas Hermann, Simon Alberti, Jochen Guck |
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| Rok vydání: | 2022 |
| Předmět: |
methods [Microscopy]
QH301-705.5 optical diffraction tomography Science Cells Intracellular Space methods [Tomography Optical] density measurement mechanical properties Physics of Living Systems Brillouin microscopy Fluorescence General Biochemistry Genetics and Molecular Biology cell biology physics of living systems Tomography Optical Humans HeLa cells human Biology (General) Cell Nucleus cytology [Cells] Microscopy General Immunology and Microbiology General Neuroscience Cell Biology General Medicine Refractometry phase transition Medicine ddc:600 Research Article Human ultrastructure [Cells] |
| Zdroj: | eLife 11, e68490 (2022). doi:10.7554/eLife.68490 eLife eLife, Vol 11 (2022) |
| ISSN: | 2050-084X |
| DOI: | 10.7554/elife.68490 |
| Popis: | 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 epifluorescence 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. |
| Databáze: | OpenAIRE |
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