Chapter 10 - Histochemical and immunohistochemical staining methods to visualize mitochondrial proteins and activity
Section snippets
Rationale
One of the first histochemical methods to detect alterations of mitochondria in skeletal muscle was the modified Gomori trichrome method of Engel and Cunningham (1963). In this method, frozen sections (6–10 μm thick) are incubated in Gill's hematoxylin for 30 min, rinsed, placed in Gomori trichrome solution for 1 h, and dipped in 0.2% acetic acid, dehydrated in graded ethanols, and cleared in xylene or xylene-substitute clearing agent to be eventually mounted with Permount. This stain results in
Rationale
Immunohistochemistry is a technique based on the detection of epitopes using antibodies, allowing for the identification of specific proteins in single cells. Immunohistochemical techniques can be used with frozen tissue or formalin-fixed paraffin-embedded tissue, which represents an important advantage compared to histochemistry techniques. Moreover, there is a great array of commercially available antibodies against both mtDNA- and nDNA-encoded subunits of the respiratory chain complex, other
Acknowledgments
Images were collected in the Confocal and Specialized Microscopy Shared Resource of the Herbert Irving Comprehensive Cancer Center at Columbia University, supported by NIH grant #P30 CA013696 (National Cancer Institute). The confocal microscope was purchased with NIH grant #S10 RR025686.
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