An Optical Technique for Mapping Microviscosity Dynamics in Cellular Organelles

作者：Chambers Joseph E; Kubankova Marketa; Huber Roland G; Lopez Duarte Ismael; Avezov Edward; Bond Peter J; Marciniak Stefan J^*; Kuimova Marina K^*

来源：ACS Nano, 2018, 12(5): 4398-4407.

DOI：10.1021/acsnano.8b00177

• microviscosity
• diffusion
• organelle
• cell biophysics
• fluorescence
• FLIM
• molecular rotors

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. ROW 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 mitochondria' matrix, ROW revealed several striking findings: a broad heterogeneity of microviscosity among individual mitochondria, unparalleled resilience to osmotic stress, and real-time changes in microviscosity during mitochondria' depolarization. These findings demonstrate the use of ROW to explore the biophysical mechanisms underlying cell biological processes.

• 出版日期2018-5
• 单位北京理工大学