TY - JOUR
T1 - A Comprehensive Approach to Sample Preparation for Electron Microscopy and the Assessment of Mitochondrial Morphology in Tissue and Cultured Cells
AU - Hinton, Antentor
AU - Katti, Prasanna
AU - Christensen, Trace A.
AU - Mungai, Margaret
AU - Shao, Jianqiang
AU - Zhang, Liang
AU - Trushin, Sergey
AU - Alghanem, Ahmad
AU - Jaspersen, Adam
AU - Geroux, Rachel E.
AU - Neikirk, Kit
AU - Biete, Michelle
AU - Lopez, Edgar Garza
AU - Shao, Bryanna
AU - Vue, Zer
AU - Vang, Larry
AU - Beasley, Heather K.
AU - Marshall, Andrea G.
AU - Stephens, Dominique
AU - Damo, Steven
AU - Ponce, Jessica
AU - Bleck, Christopher K.E.
AU - Hicsasmaz, Innes
AU - Murray, Sandra A.
AU - Edmonds, Ranthony A.C.
AU - Dajles, Andres
AU - Koo, Young Do
AU - Bacevac, Serif
AU - Salisbury, Jeffrey L.
AU - Pereira, Renata O.
AU - Glancy, Brian
AU - Trushina, Eugenia
AU - Abel, E. Dale
N1 - Publisher Copyright:
© 2023 The Authors. Advanced Biology published by Wiley-VCH GmbH.
PY - 2023/10
Y1 - 2023/10
N2 - Mitochondria respond to metabolic demands of the cell and to incremental damage, in part, through dynamic structural changes that include fission (fragmentation), fusion (merging of distinct mitochondria), autophagic degradation (mitophagy), and biogenic interactions with the endoplasmic reticulum (ER). High resolution study of mitochondrial structural and functional relationships requires rapid preservation of specimens to reduce technical artifacts coupled with quantitative assessment of mitochondrial architecture. A practical approach for assessing mitochondrial fine structure using two dimensional and three dimensional high-resolution electron microscopy is presented, and a systematic approach to measure mitochondrial architecture, including volume, length, hyperbranching, cristae morphology, and the number and extent of interaction with the ER is described. These methods are used to assess mitochondrial architecture in cells and tissue with high energy demand, including skeletal muscle cells, mouse brain tissue, and Drosophila muscles. The accuracy of assessment is validated in cells and tissue with deletion of genes involved in mitochondrial dynamics.
AB - Mitochondria respond to metabolic demands of the cell and to incremental damage, in part, through dynamic structural changes that include fission (fragmentation), fusion (merging of distinct mitochondria), autophagic degradation (mitophagy), and biogenic interactions with the endoplasmic reticulum (ER). High resolution study of mitochondrial structural and functional relationships requires rapid preservation of specimens to reduce technical artifacts coupled with quantitative assessment of mitochondrial architecture. A practical approach for assessing mitochondrial fine structure using two dimensional and three dimensional high-resolution electron microscopy is presented, and a systematic approach to measure mitochondrial architecture, including volume, length, hyperbranching, cristae morphology, and the number and extent of interaction with the ER is described. These methods are used to assess mitochondrial architecture in cells and tissue with high energy demand, including skeletal muscle cells, mouse brain tissue, and Drosophila muscles. The accuracy of assessment is validated in cells and tissue with deletion of genes involved in mitochondrial dynamics.
KW - automated serial block-face SEM
KW - focused ion beam SEM
KW - mitochondria-endoplasmic reticulum communication
KW - mitochondrial dynamics
KW - mitochondrial morphology
KW - serial-section TEM
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U2 - 10.1002/adbi.202200202
DO - 10.1002/adbi.202200202
M3 - Article
C2 - 37140138
AN - SCOPUS:85158043967
SN - 2701-0198
VL - 7
JO - Advanced Biology
JF - Advanced Biology
IS - 10
M1 - 2200202
ER -