Protocol for measuring mitochondrial size in mouse and human liver tissues

Balamurugan Ramatchandirin; Miho Iijima; Arisa Ikeda; Alexia Pearah; Sally Radovick; Fredric E Wondisford; Hiromi Sesaki; Ling He

doi:10.1016/j.xpro.2024.102842

Protocol for measuring mitochondrial size in mouse and human liver tissues

STAR Protoc. 2024 Mar 15;5(1):102842. doi: 10.1016/j.xpro.2024.102842. Epub 2024 Jan 18.

Authors

Balamurugan Ramatchandirin¹, Miho Iijima², Arisa Ikeda², Alexia Pearah¹, Sally Radovick³, Fredric E Wondisford⁴, Hiromi Sesaki⁵, Ling He⁶

Affiliations

¹ Departments of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
² Departments of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
³ Institute of Clinical Translational Sciences, University of Arizona, Tucson, AZ 85721, USA.
⁴ Departments of Internal Medicine, University of Arizona College of Medicine-Phoenix, Phoenix, AZ 85004, USA.
⁵ Departments of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: hsesaki@jhmi.edu.
⁶ Departments of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Departments of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: heling@jhmi.edu.

Abstract

Mitochondrial dynamic process is important for cell viability, metabolic activity, and mitochondria health. Here, we present a protocol for measuring mitochondrial size through immunofluorescence staining, confocal imaging, and analysis in ImageJ. We describe the steps for tissue processing, antigen retrieval, mitochondrial staining using an integrating immunofluorescence assay, and computerized image analysis to measure each mitochondrial size in mouse and human liver tissues. This protocol reduces tissue sample volume and processing time for the preparation of primary cells. For complete details on the use and execution of this protocol, please refer to Pearah et al.¹.

Keywords: Cell Biology; Health Sciences; Metabolism.

MeSH terms

Animals
Cell Survival
Humans
Image Processing, Computer-Assisted*
Liver*
Mice
Mitochondria
Mitochondrial Size

Abstract

MeSH terms

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