Imaging Mitochondrial Hydrogen Peroxide in Living Cells

Hydrogen peroxide (H2O2) produced from mitochondria is intimately involved in human health and disease but is challenging to selectively monitor inside living systems. The fluorescent probe MitoPY1 provides a practical tool for imaging mitochondrial H2O2 and has been demonstrated to function in a variety of diverse cell types. In this chapter, we describe the synthetic preparation of the small-molecule probe MitoPY1, methods for validating this probe in vitro and in live cells, and an example procedure for measuring mitochondrial H2O2 in a cell culture model of Parkinson’s disease.