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Shadow Electrochemiluminescence Microscopy of Single Mitochondria
Author(s) -
Ma Yumeng,
Colin Camille,
Descamps Julie,
Arbault Stéphane,
Sojic Neso
Publication year - 2021
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202105867
Subject(s) - mitochondrion , fluorescence microscope , photobleaching , microscopy , chemistry , electrochemiluminescence , biophysics , organelle , shadow (psychology) , fluorescence , biochemistry , biology , electrode , optics , psychology , physics , psychotherapist
Abstract Mitochondria are the subcellular bioenergetic organelles. The analysis of their morphology and topology is essential to provide useful information on their activity and metabolism. Herein, we report a label‐free shadow electrochemiluminescence (ECL) microscopy based on the spatial confinement of the ECL‐emitting reactive layer to image single living mitochondria deposited on the electrode surface. The ECL mechanism of the freely‐diffusing [Ru(bpy) 3 ] 2+ dye with the sacrificial tri‐n‐propylamine coreactant restrains the light‐emitting region to a micrometric thickness allowing to visualize individual mitochondria with a remarkable sharp negative optical contrast. The imaging approach named “shadow ECL” (SECL) reflects the negative imprint of the local diffusional hindrance of the ECL reagents by each mitochondrion. The statistical analysis of the colocalization of the shadow ECL spots with the functional mitochondria revealed by classical fluorescent biomarkers, MitoTracker Deep Red and the endogenous intramitochondrial NADH, validates the reported methodology. The versatility and extreme sensitivity of the approach are further demonstrated by visualizing single mitochondria, which remain hardly detectable with the usual biomarkers. Finally, by alleviating problems of photobleaching and phototoxicity associated with conventional microscopy methods, SECL microscopy should find promising applications in the imaging of subcellular structures.