The mitochondria-associated endoplasm reticulum (mito-ER) membrane plays an important role in calcium flux, autophagy, lipid exchange, the apoptotic commencement mechanism, and ER control over mitochondrial biogenesis and dynamics. Due to complex intercellular functions, these organelles more often reside close to one another at nanometer-scale dimensions. Herein, we present an approach based on 3D superlocalization of plasmon nanoparticles (NPs) along with dual-view enhanced dark-field (EDF) microscopy to detect mito-ER contacts concurrently at real-time. 10-nm thick slicing in the axial direction provided 10-nm spatial resolution in 3D for plasmonic NPs in single living cells. To analyze of nanotag-labeled mitochondria and ER with concurrent imaging at two wavelengths, a dual-view approach was implemented alongside EDF microscopy. 20-nm silver NP (AgNP)-labeled mitochondria and 12-nm gold NP (AuNP)-labeled ER in living HeLa cells collected with the dual-wavelength, synchronized imaging system. After applying the superlocalization method, the relative positions of the mitochondria and ER in mito-ER contacts were resolved. The distances between mitochondria and the ER were quantified, typically as 45.3 nm. These results indicate that this method was a reliable and convenient approach for investigating the real-time concurrent intracellular multi-organelle super-localization.<br/>

Multiorganelle concurrent imaging in single cells by 3D superlocalization of dual-code enhanced dark-field microscopy

2019 Fall National Meeting

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.