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Here we use the combination of fluorescent molecular rotors (FMR) and fluorescence lifetime imaging (FLIM) to image matrix viscosity in live cells. We use non-cancerous, epithelial human cornea (HCE) cells as a model system. We find that mitochondrial matrix viscosity varies between individual cells and even between individual organelles, showcasing the potential of viscosity imaging for cell biology purposes.

Imaging mitochondrial matrix viscosity in live cells via fluorescence lifetime imaging (FLIM) of fluorescent molecular rotors