Live cell mitochondrial 3-dimensional dynamic ultrastructures under oxidative phosphorylation revealed by a Pyridine-BODIPY probe

De Novo Designed Self-Assembling Rhodamine Probe for Real-Time, Long-Term and Quantitative Live-Cell Nanoscopy

“…Our probe quantifies mitochondrial cristae change under stimuli using the self-labeling protein tag approach with STED imaging. Compared with the existing methods to quantify mitochondrial cristae, − our approach provides higher resolution than methods using probes with structured illumination microscopy (SIM). It also gives more reliable images than methods using membrane potential-dependent probes.…”

Xanthene, cyanine, oxazine and BODIPY: the four pillars of the fluorophore empire for super-resolution bioimaging

“…25(A)). 215 Py-BODIPY revealed interference-free upright targeting ability towards mitochondrial membranes in living cells. The best part of this rationally designed probe is its membrane potential independent highly specific mitochondrial membrane targeting ability, which allowed the robust staining of the specific organelle in both live-cell and fixed cell samples.…”

“…Therefore, the BODIPY-based newly designed fluorescent probe Py-BODIPY was successfully utilized in STED nanoscopy not only to obtain the nanoscopic images of mitochondria but also to observe the dynamic mitochondrial nano-structures with sub-diffraction resolution which clearly demonstrated the time-dependant evolution of the mitochondrial 3D ultrastructure under oxidative phosphorylation conditions. 215…”

“…Therefore, the BODIPY-based newly designed fluorescent probe Py-BODIPY was successfully utilized in STED nanoscopy This journal is © The Royal Society of Chemistry 2023 not only to obtain the nanoscopic images of mitochondria but also to observe the dynamic mitochondrial nano-structures with sub-diffraction resolution which clearly demonstrated the time-dependant evolution of the mitochondrial 3D ultrastructure under oxidative phosphorylation conditions. 215 Mitochondrial membrane-bound VDPs are believed to have close relations with some pathogenic events, even though the exact structural and functional details are not realized properly. 216 Since a significant portion of mitochondrial VDPs exist in the mitochondrial membrane, a rationally designed STED compatible probe that can target the VDPs in the mitochondrial membrane would be highly crucial to elucidate the structural and functional details of VDP dynamics at the nanoscale level.…”

Small-molecule fluorogenic probes for mitochondrial nanoscale imaging

“…105 In 2021, Zhu et al developed a fluorogenic probe, Py-BODIPY, which could target mitochondrial membranes in live cells without disturbing their physiological microenvironment. 106 This probe binds with mitochondria by its protonated pyridine moiety and the formation of N-S bonds with the mitochondrial membrane protein independent of the MMP. Its crista specificity, low molecular weight, high quantum yield (F F = 0.35) and good photostability allow the observation of mitochondrial dynamic nanostructures with unprecedented resolution, while 3D information is represented by ''depth-coded'' bars (Fig.…”