Chambers, Joseph E.Kubánková, MarkétaHuber, Roland G.López Duarte, IsmaelAvezov, EdwardBond, Peter J.Marciniak, Stefan J.Kuimova, Marina K.2025-01-222025-01-222018-04-12Chambers 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.8b001771936-08511936-086X10.1021/acsnano.8b00177https://hdl.handle.net/20.500.14352/115607Microscopic 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.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal articlehttps://doi.org/10.1021/acsnano.8b00177open access615.31615:54microviscositydiffusionorganellecell biophysicsfluorescenceFLIMmolecular rotorsQuímica farmaceútica23 Química