Protocol for measuring mitochondrial size in mouse and human liver tissues

• Published in: STAR protocols Vol. 5; no. 1; p. 102842
• Main Authors: Ramatchandirin, Balamurugan, Iijima, Miho, Ikeda, Arisa, Pearah, Alexia, Radovick, Sally, Wondisford, Fredric E., Sesaki, Hiromi, He, Ling
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.03.2024; Elsevier
• Subjects: Animals; Cell Biology; Cell Survival; Health Sciences; Humans; Image Processing, Computer-Assisted; Liver; Metabolism; Mice; Mitochondria; Mitochondrial Size; Protocol; Metabolism; Cell Biology; Health Sciences
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.102842

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 [Display omitted] •Protocol for measuring mitochondrial size in mouse and human liver tissues•Detailed step for mitochondrial staining using immunofluorescence assays•Use high-pressure cooker for antigen retrieval to achieve high-quality images•Quantify individual mitochondrial size using ImageJ software Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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 of 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.