Visualization and quantification of cardiac mitochondrial protein clusters with STED microscopy

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• 作者：Harpreet Singh^a ; Rong Lu^a ; Pedro Felipe Gardeazá ; bal Rodrí ; guez^a ; Yong Wu^a ; Jean Chrisostome Bopassa^a ; Enrico Stefani^a ; ^b ; ^d ; ¹ ; Ligia Toro^a ; ^c ; ^d ; ¹ ; ^{ltoro@ucla.edu}
• 关键词：STED ; STimulated Emission Depletion ; VDAC1 ; Voltage Dependent Anion Channel 1 ; Cox2 ; Cytochrome c Oxidase subunit 2 ; SFKs ; Src family of tyrosine kinases ; PPI ; protein proximity index ; FWHM ; Full Width Half Maximum ; TDE ; 2 ; 2&prime ; -Thiodiethanol
• 刊名：Mitochondrion
• 出版年：2012
• 期刊代码：39_15677249
• 类别：neu
• 出版时间：March, 2012
• 卷：12
• 期：2
• 页码：230-236
• 文件大小：1040 K

摘要

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.