Long-Term Dynamic Imaging of Cellular Processes Using an AIE Lipid Order Probe in the Dual-Color Mode

Lipid order in the cytoplasm membrane of eukaryotic cells undergoes dynamic changes in almost all cellular processes. Dynamically monitoring these changes is of essential biological significance and remains challenging. This work provides the first aggregation-induced emission probe, TPNPDA-C15, with highly three-dimensional specificity to cell membranes for fluorescent imaging of lipid order of live cells. TPNPDA-C15 displays red fluorescence enhancement with the viscosity increase while emits yellow fluorescence when aggregates form. Imaging analyses of giant unilamellar vesicles and live cells under osmotic shock by the probe demonstrate its sensitive response to the degree of phospholipids packing on artificial and cell membranes. Taking advantage of its superior low photocytotoxicity and high photostability, TPNPDA-C15 is further applied for long-term dynamic imaging of entire live cell physiological processes including apoptosis, ferroptosis, and mitosis in the dual-color mode. With the analysis of fluorescence signal changes in the two fluorescence channels, TPNPDA-C15 serves as a robust fluorescent probe for the imaging study of cellular dynamics.