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Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy.

Mitochondrion (2011-10-11)
Harpreet Singh, Rong Lu, Pedro Felipe Gardeazábal Rodríguez, Yong Wu, Jean Chrisostome Bopassa, Enrico Stefani, Ligia Toro
ABSTRACT

The visualization and quantification of mitochondria-associated proteins with high power microscopy methods is of particular interest to investigate protein architecture in this organelle. We report the usage of a custom-made STimulated Emission Depletion (STED) fluorescence nanoscope with ~30nm lateral resolution for protein mapping of Percoll-purified viable mitochondria from murine heart. Using this approach, we were able to quantify and resolve distinct protein clusters within mitochondria; specifically, cytochrome c oxidase subunit 2 is distributed in clusters of ~28nm; whereas the voltage dependent anion channel 1 displays three size distributions of ~33, ~55 and ~83nm.

MATERIALS
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