Collection of Isolated Cells for Studying Mitochondrial DNA Mutations Within Individual Cells

Abstract

Mitochondrial genome integrity is an important issue in somatic mitochondrial genetics. Development of quantitative methods is indispensable to somatic mitochondrial genetics as quantitative studies are required to characterize heteroplasmy and mutation processes, as well as their effects on phenotypic developments. This chapter outlines the methods for collecting individual cells appropriate for analysis of mtDNA mutations by single-molecule PCR. In addition, we describe the protocols for respiratory complexes II and IV in these cells. Together, the identification of respiratory deficiency and mtDNA mutations within single cells provides a powerful means of evaluating the importance of these events in disease and aging.

A novel and effective separation method for single mitochondria analysis

“…In mitochondrial research, this technique has been used to analyse mtDNA of cancer tissue (e.g. Aldridge et al, 2003), and of individual cells (Kraytsberg et al, 2009).…”

Mitochondrial DNA deletions in mice in men: Substantia nigra is much less affected in the mouse

“…The area containing pigmented neurons was scraped from the section saturated with glycerol for better visibility as described elsewhere [16]. In the mouse, substantia nigra was identified by its position in the midbrain by inspecting a large series of frozen sections of the entire mouse brain.…”