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Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

Authors
Jung, MinkyoMun, Ji Young
Issue Date
Jul-2019
Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
Keywords
Biochemistry; Issue 149; mitochondria; ER; APEX; HRP; correlative light and electron microscopy; 3DEM
Citation
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no.149
Journal Title
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Number
149
URI
http://scholarworks.bwise.kr/kbri/handle/2023.sw.kbri/683
DOI
10.3791/59750
ISSN
1940-087X
Abstract
Cellular organelles, such as mitochondria and endoplasmic reticulum (ER), create a network to perform a variety of functions. These highly curved structures are folded into various shapes to form a dynamic network depending on the cellular conditions. Visualization of this network between mitochondria and ER has been attempted using super-resolution fluorescence imaging and light microscopy; however, the limited resolution is insufficient to observe the membranes between the mitochondria and ER in detail. Transmission electron microscopy provides good membrane contrast and nanometer-scale resolution for the observation of cellular organelles; however, it is exceptionally time-consuming when assessing the three-dimensional (3D) structure of highly curved organelles. Therefore, we observed the morphology of mitochondria and ER via correlative light-electron microscopy (CLEM) and volume electron microscopy techniques using enhanced ascorbate peroxidase 2 and horseradish peroxidase staining. An en bloc staining method, ultrathin serial sectioning (array tomography), and volume electron microscopy were applied to observe the 3D structure. In this protocol, we suggest a combination of CLEM and 3D electron microscopy to perform detailed structural studies of mitochondria and ER.
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연구본부 (신경회로 연구그룹)
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