Targeted
Photoconvertible BODIPYs Based on Directed
Photooxidation-Induced Conversion for Applications in Photoconversion
and Live Super-Resolution Imaging
posted on 2024-06-11, 16:34authored byLazare Saladin, Victor Breton, Valentine Le Berruyer, Paul Nazac, Thiebault Lequeu, Pascal Didier, Lydia Danglot, Mayeul Collot
Photomodulable
fluorescent probes are drawing increasing attention
due to their applications in advanced bioimaging. Whereas photoconvertible
probes can be advantageously used in tracking, photoswitchable probes
constitute key tools for single-molecule localization microscopy to
perform super-resolution imaging. Herein, we shed light on a red and
far-red BODIPY, namely, BDP-576 and BDP-650, which possess both properties
of conversion and switching. Our study demonstrates that these pyrrolyl-BODIPYs
convert into typical green- and red-emitting BODIPYs that are perfectly
adapted to microscopy. We also showed that this pyrrolyl-BODIPYs undergo
Directed Photooxidation Induced Conversion, a photoconversion mechanism
that we recently introduced, where the pyrrole moiety plays a central
role. These unique features were used to develop targeted photoconvertible
probes toward different organelles or subcellular units (plasma membrane,
mitochondria, nucleus, actin, Golgi apparatus, etc.) using chemical targeting moieties and a Halo tag. We notably showed
that BDP-650 could be used to track intracellular vesicles over more
than 20 min in two-color imagings with laser scanning confocal microscopy,
demonstrating its robustness. The switching properties of these photoconverters
were studied at the single-molecule level and were then successfully
used in live single-molecule localization microscopy in epithelial
cells and neurons. Both membrane- and mitochondria- targeted probes
could be used to decipher membrane 3D architecture and mitochondrial
dynamics at the nanoscale. This study builds a bridge between the
photoconversion and photoswitching properties of probes undergoing
directed photooxidation and shows the versatility and efficacy of
this mechanism in advanced live imaging.