Preparation of "functional" mitochondria: a challenging business

As the powerhouse of the cell, mitochondria play a crucial role in many aspects of life, whereby mitochondrial dysfunctions are associated with pathogenesis of many diseases, like neurodegenerative diseases, obesity, cancer, and metabolic as well as cardiovascular disorders. Mitochondria analysis frequently starts with isolation and enrichment procedures potentially affecting mitochondrial morphology having impact on their function. Due to the complex mitochondrial morphology, the major task is to preserve their structural integrity. Here we critically review a commonly used isolation procedure for mitochondria utilizing differential (gradient) centrifugation and depict major challenges to achieve "functional" mitochondria as basis for comprehensive physiological studies.