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Chambers, Joseph E. Kubánková, Markéta Huber, Roland G. López Duarte, Ismael Avezov, Edward Bond, Peter J. Marciniak, Stefan J. Kuimova, Marina K. 2025-01-22T14:06:44Z 2025-01-22T14:06:44Z 2018-04-12 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 1936-0851 1936-086X 10.1021/acsnano.8b00177 https://hdl.handle.net/20.500.14352/115607 https://doi.org/10.1021/acsnano.8b00177 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. eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 International An Optical Technique for Mapping Microviscosity Dynamics in Cellular Organelles journal article