Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy

Harpreet Singh; Rong Lu; Pedro Felipe Gardeazábal Rodríguez; Yong Wu; Jean Chrisostome Bopassa; Enrico Stefani; Ligia Toro

doi:10.1016/j.mito.2011.09.004

Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy

Harpreet Singh
, Rong Lu
, Pedro Felipe Gardeazábal Rodríguez
, Yong Wu
, Jean Chrisostome Bopassa
, Enrico Stefani
, Ligia Toro

Research output: Contribution to journal › Article › peer-review

52 Scopus citations

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 ~. 30. nm 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 ~. 28. nm; whereas the voltage dependent anion channel 1 displays three size distributions of ~. 33, ~. 55 and ~. 83. nm.

Original language	English (US)
Pages (from-to)	230-236
Number of pages	7
Journal	Mitochondrion
Volume	12
Issue number	2
DOIs	https://doi.org/10.1016/j.mito.2011.09.004
State	Published - Mar 2012
Externally published	Yes

Keywords

Cox2
Imaging
Mitochondria
STED
Subdiffraction-resolution
VDAC1

ASJC Scopus subject areas

Molecular Medicine
Molecular Biology
Cell Biology

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10.1016/j.mito.2011.09.004

Cite this

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title = "Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy",

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 \textasciitilde{}. 30. nm 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 \textasciitilde{}. 28. nm; whereas the voltage dependent anion channel 1 displays three size distributions of \textasciitilde{}. 33, \textasciitilde{}. 55 and \textasciitilde{}. 83. nm.",

keywords = "Cox2, Imaging, Mitochondria, STED, Subdiffraction-resolution, VDAC1",

author = "Harpreet Singh and Rong Lu and Rodr{\'i}guez, \{Pedro Felipe Gardeaz{\'a}bal\} and Yong Wu and Bopassa, \{Jean Chrisostome\} and Enrico Stefani and Ligia Toro",

note = "Funding Information: We thank Dr. Jeff Abramson (UCLA) for VDAC1 recombinant protein. This work was supported by NIH grants, HL54970 (LT) and HL088640 (ES) and the American Heart Association Fellowships 09POST2190008 (JCB) and 10POST4230081 (YW) and National Scientist Development Grant 11SDG7230059 (H.S.).",

year = "2012",

month = mar,

doi = "10.1016/j.mito.2011.09.004",

language = "English (US)",

volume = "12",

pages = "230--236",

journal = "Mitochondrion",

issn = "1567-7249",

publisher = "Elsevier B.V.",

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AU - Singh, Harpreet

AU - Lu, Rong

AU - Rodríguez, Pedro Felipe Gardeazábal

AU - Wu, Yong

AU - Bopassa, Jean Chrisostome

AU - Stefani, Enrico

AU - Toro, Ligia

N1 - Funding Information: We thank Dr. Jeff Abramson (UCLA) for VDAC1 recombinant protein. This work was supported by NIH grants, HL54970 (LT) and HL088640 (ES) and the American Heart Association Fellowships 09POST2190008 (JCB) and 10POST4230081 (YW) and National Scientist Development Grant 11SDG7230059 (H.S.).

PY - 2012/3

Y1 - 2012/3

N2 - 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 ~. 30. nm 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 ~. 28. nm; whereas the voltage dependent anion channel 1 displays three size distributions of ~. 33, ~. 55 and ~. 83. nm.

AB - 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 ~. 30. nm 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 ~. 28. nm; whereas the voltage dependent anion channel 1 displays three size distributions of ~. 33, ~. 55 and ~. 83. nm.

KW - Cox2

KW - Imaging

KW - Mitochondria

KW - STED

KW - Subdiffraction-resolution

KW - VDAC1

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UR - https://www.scopus.com/pages/publications/84858006645#tab=citedBy

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DO - 10.1016/j.mito.2011.09.004

M3 - Article

C2 - 21982778

AN - SCOPUS:84858006645

SN - 1567-7249

VL - 12

SP - 230

EP - 236

JO - Mitochondrion

JF - Mitochondrion

IS - 2

ER -

Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy

Abstract

Keywords

ASJC Scopus subject areas

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