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Significance Stimulated emission depletion (STED) microscopy is one of the most appealing tools to visualize nanoscale cellular structures and dynamics in living cells. However, its practical utility is significantly limited by the rapid photobleaching of fluorescent dyes under the ultrastrong depletion laser. In this context, superphotostable fluorescent probes, which enable repeated recording of the STED images, are crucial chemical tools to extend the application of STED microscopy in live-cell imaging. Herein, we report a developed fluorescent probe, MitoPB Yellow, which has the characters of outstanding photostability, long fluorescence lifetime, and mitochondrial inner membrane selectivity for staining. In combination with a time-gated-STED microscopy, the mitochondrial inner-membrane structures and dynamics can be impressively visualized in living cells.

A photostable fluorescent marker for the superresolution live imaging of the dynamic structure of the mitochondrial cristae