A Universal Approach to Analyzing Transmission Electron Microscopy with ImageJ

Jacob Lam; Prasanna Katti; Michelle Biete; Margaret Mungai; Salma AshShareef; Kit Neikirk; Edgar Garza Lopez; Zer Vue; Trace A Christensen; Heather K Beasley; Taylor A Rodman; Sandra A Murray; Jeffrey L Salisbury; Brian Glancy; Jianqiang Shao; Renata O Pereira; E Dale Abel; Antentor Hinton Jr

doi:10.3390/cells10092177

A Universal Approach to Analyzing Transmission Electron Microscopy with ImageJ

Cells. 2021 Aug 24;10(9):2177. doi: 10.3390/cells10092177.

Authors

Jacob Lam^{1

2}, Prasanna Katti³, Michelle Biete⁴, Margaret Mungai⁵, Salma AshShareef^{1

2}, Kit Neikirk⁴, Edgar Garza Lopez⁶, Zer Vue⁶, Trace A Christensen⁷, Heather K Beasley⁶, Taylor A Rodman⁶, Sandra A Murray⁸, Jeffrey L Salisbury^{7

9}, Brian Glancy^{3

10}, Jianqiang Shao¹¹, Renata O Pereira^{1

2}, E Dale Abel^{1

2}, Antentor Hinton Jr^{6

7}

Affiliations

¹ Department of Internal Medicine, Carver College of Medicine, University of Iowa, 375 Newton Rd, Iowa City, IA 52242, USA.
² Fraternal Order of Eagles Diabetes Research Center, University of Iowa, 375 Newton Rd, Iowa City, IA 52242, USA.
³ National Heart, Lung and Blood Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA.
⁴ Daniel K. Inouye College of Pharmacy, University of Hawaii at Hilo, 200 West Kawili St, Hilo, HI 96720, USA.
⁵ Department of Molecular and Cell Biology, University of California Berkeley, 142 Weill Hall, Berkeley, CA 94720, USA.
⁶ Department of Molecular Physiology and Biophysics, Vanderbilt University, 2201 West End Ave, Nashville, TN 37235, USA.
⁷ Microscopy and Cell Analysis Core Facility, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
⁸ Department of Cell Biology, School of Medicine, University of Pittsburgh, 3550 Terrace St., Pittsburgh, PA 15213, USA.
⁹ Department of Biochemistry and Molecular Biology, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
¹⁰ National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, 9000 Rockville Pike, Bethesda, MD 20892, USA.
¹¹ Central Microscopy Research Facility, University of Iowa, Iowa City, IA 52242, USA.

Abstract

Transmission electron microscopy (TEM) is widely used as an imaging modality to provide high-resolution details of subcellular components within cells and tissues. Mitochondria and endoplasmic reticulum (ER) are organelles of particular interest to those investigating metabolic disorders. A straightforward method for quantifying and characterizing particular aspects of these organelles would be a useful tool. In this protocol, we outline how to accurately assess the morphology of these important subcellular structures using open source software ImageJ, originally developed by the National Institutes of Health (NIH). Specifically, we detail how to obtain mitochondrial length, width, area, and circularity, in addition to assessing cristae morphology and measuring mito/endoplasmic reticulum (ER) interactions. These procedures provide useful tools for quantifying and characterizing key features of sub-cellular morphology, leading to accurate and reproducible measurements and visualizations of mitochondria and ER.

Keywords: ImageJ; Mitochondria Endoplasmic Reticulum Contacts (MERCs); TEM analysis; TEM quantification; cristae; image analysis; image processing; mitochondria.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cells, Cultured
Endoplasmic Reticulum / physiology
Male
Mice
Mice, Inbred C57BL
Microscopy, Electron, Transmission / methods*
Mitochondria / physiology
Mitochondrial Membranes / physiology
Software

Abstract

Publication types

MeSH terms

Grants and funding