Histochemical and immunohistochemical staining methods to visualize mitochondrial proteins and activity

Methods Cell Biol. 2020:155:247-270. doi: 10.1016/bs.mcb.2019.11.024. Epub 2020 Jan 20.


We describe here reliable histochemical and immunohistochemical techniques to visualize mitochondria and respiratory chain dysfunction in tissue sections. These morphological methods have been widely used for years, and yet remain relevant to obtain insight into the pathogenesis of mitochondrial diseases. Today, mitochondrial medicine is changing rapidly and genetic information plays an increasing role in the diagnostic process, owing to advances in next-generation sequencing. However, tissue analysis and morphological categorization remain essential, especially when genetic abnormalities of unknown significance might complicate a diagnostic odyssey. Furthermore, tissue assessment is an essential step in laboratory investigation using animal or cell models, in order to assess the distribution, severity, and/or progression of the disease, and to evaluate the effects of possible treatments. Optimized and reproducible staining and imaging methodology are the key elements for accurate tissue assessment. When these methods are used properly and integrated with wisely chosen genetic and biochemical approaches, powerful information can be obtained about the structure and function of mitochondria in both animal model systems and human patients. While the described protocols refer to skeletal muscle and brain mitochondria, the methods described can be applied to any tissue type.

Keywords: Cytochrome c oxidase; Histochemistry; Immunofluorescence; Immunohistochemistry; Muscle; Succinate dehydrogenase.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Histocytochemistry / methods*
  • Humans
  • Mitochondrial Proteins / metabolism*
  • Paraffin Embedding
  • Staining and Labeling / methods*
  • Tissue Fixation


  • Mitochondrial Proteins