Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy |
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Authors: | Singh Harpreet Lu Rong Rodríguez Pedro Felipe Gardeazábal Wu Yong Bopassa Jean Chrisostome Stefani Enrico Toro Ligia |
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Institution: | Department of Anesthesiology, Division of Molecular Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA. |
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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. |
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