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Mechanosensitive Fluorescent Probes to Image Membrane Tension in Mitochondria, Endoplasmic Reticulum, and Lysosomes
- Goujon, Antoine, Colom, Adai, Straková, Karolína, Mercier, Vincent, Mahecic, Dora, Manley, Suliana, Sakai, Naomi, Roux, Aurélien, Matile, Stefan
- Journal of the American Chemical Society 2019 v.141 no.8 pp. 3380-3384
- endocytosis, endoplasmic reticulum, fluorescence, fluorescent dyes, image analysis, lysosomes, microscopy, mitochondria, osmotic stress, plasma membrane
- Measuring forces inside cells is particularly challenging. With the development of quantitative microscopy, fluorophores which allow the measurement of forces became highly desirable. We have previously introduced a mechanosensitive flipper probe, which responds to the change of plasma membrane tension by changing its fluorescence lifetime and thus allows tension imaging by FLIM. Herein, we describe the design, synthesis, and evaluation of flipper probes that selectively label intracellular organelles, i.e., lysosomes, mitochondria, and the endoplasmic reticulum. The probes respond uniformly to osmotic shocks applied extracellularly, thus confirming sensitivity toward changes in membrane tension. At rest, different lifetimes found for different organelles relate to known differences in membrane organization rather than membrane tension and allow colabeling in the same cells. At the organelle scale, lifetime heterogeneity provides unprecedented insights on ER tubules and sheets, and nuclear membranes. Examples on endosomal trafficking or increase of tension at mitochondrial constriction sites outline the potential of intracellularly targeted fluorescent tension probes to address essential questions that were previously beyond reach.