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Le Berruyer, V.

Publications and source records attributed to Le Berruyer, V..

2 recordsLinked to original sources

Targeted Photoactivatable Green-Emitting BODIPY Based on Directed Photooxidation Induced Activation and its Application to Live Dynamic Super-Resolution Microscopy

Photoactivatable fluorescent probes are valuable tools in bioimaging for tracking cells down to single molecules and for single molecule localization microscopy. For the latter application, green emitting dyes are in demand. We herein developed an efficient green-emitting photoactivatable furanyl-BODIPY (PFB) and we established a new mechanism of photoactivation called Directed Photooxidation Induced Activation (DPIA) where the furan is photo-oxidized in a directed manner by the singlet oxygen produced by the probe. The efficient photoconverter (93-fold fluorescence enhancement at 510 nm, 49% yield conversion) is functionalizable and allowed targeting of several subcellular structures and organelles, which were photoactivated in live cells. Finally, we demonstrated the potential of PFB in super-resolution imaging by performing PhotoActivated Localization Microscopy (PALM) in live cells.

cell biology↗

Photoactivatable Plasma Membrane Probe Through Self-Triggered Photooxidation Cascade for Live Super-Resolution Microscopy

Super-resolution imaging based on the localization of single emitters requires a spatio-temporal control of the ON and OFF state. To this end, photoactivatable fluorophores are adapted as they can be turned on upon light irradiation. Here we present a concept called Self-Triggered Photooxidation Cascade (STPC) based on the photooxidation of a plasma membrane targeted leuco-rhodamine (LRhod-PM), a non-fluorescent reduced form of a rhodamine probe. Upon visible light irradiation the small number of oxidized rhodamines, Rhod-PM, acts as a photosensitizer to generate singlet oxygen capable to oxidize the OFF state LRhod-PM. We showed that this phenomenon is kinetically favored by a high local concentration and propagates quickly when the probe is embedded in membrane bilayers. In addition, we showed that the close proximity of the dyes favors the photobleaching. At the single-molecule level, the concomitant activation/bleaching phenomena allow reaching a single-molecule blinking regime enabling single-molecule localization microscopy for super-resolution of live cellular membranes.

cell biology↗