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oai:docta.ucm.es:20.500.14352/1156072025-03-18T13:57:39Zcom_20.500.14352_14col_20.500.14352_15

An Optical Technique for Mapping Microviscosity Dynamics in Cellular Organelles Chambers, Joseph E. Kubánková, Markéta Huber, Roland G. López Duarte, Ismael Avezov, Edward Bond, Peter J. Marciniak, Stefan J. Kuimova, Marina K. 615.31 615:54 microviscosity diffusion organelle cell biophysics fluorescence FLIM molecular rotors Química farmaceútica 23 Química Microscopic viscosity (microviscosity) is a key determinant of diffusion in the cell and defines the rate of biological processes occurring at the nanoscale, including enzyme-driven metabolism and protein folding. Here we establish a rotor-based organelle viscosity imaging (ROVI) methodology that enables real-time quantitative mapping of cell microviscosity. This approach uses environment-sensitive dyes termed molecular rotors, covalently linked to genetically encoded probes to provide compartment-specific microviscosity measurements via fluorescence lifetime imaging. ROVI visualized spatial and temporal dynamics of microviscosity with suborganellar resolution, reporting on a microviscosity difference of nearly an order of magnitude between subcellular compartments. In the mitochondrial matrix, ROVI revealed several striking findings: a broad heterogeneity of microviscosity among individual mitochondria, unparalleled resilience to osmotic stress, and real-time changes in microviscosity during mitochondrial depolarization. These findings demonstrate the use of ROVI to explore the biophysical mechanisms underlying cell biological processes. Alpha-1 Foundation Engineering & Physical Sciences Research Council Depto. de Química en Ciencias Farmacéuticas Fac. de Farmacia TRUE pub 2025-01-22T14:06:44Z 2025-01-22T14:06:44Z 2018-04-12 journal article VoR https://hdl.handle.net/20.500.14352/115607 1936-0851 1936-086X 10.1021/acsnano.8b00177 eng Chambers JE, Kubánková M, Huber RG, López-Duarte I, Avezov E, Bond PJ, et al. An optical technique for mapping microviscosity dynamics in cellular organelles. ACS Nano [Internet]. 22 de mayo de 2018 [citado 22 de enero de 2025];12(5):4398-407. Disponible en: https://pubs.acs.org/doi/10.1021/acsnano.8b00177 Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ open access application/pdf American Chemical Society