What processes are involved in mitochondrial isolation from cells and tissues?

Mitochondrial isolation protocols consist of two general steps. 

1. The first step is the cell disruption to rupture the cells and release their structures via homogenization. This can be done using a  buffer or mechanical homogenizer to disrupt the cell membranes. The second step involves a type of isolation technique (e.g. differential centrifugation, density gradient centrifugation) to collect fractions that are highly concentrated with mitochondria. Differential centrifugation (DC) is a widely used technique in cell biology for separating different cellular components based on their sedimentation coefficients, which are influenced by density and shape. DC methods involve a two-step centrifugation process. The first step is performed at low speeds to eliminate intact cells, nuclei, and debris, effectively clearing the whole cell extract. 
2. The second step involves high-speed centrifugation, enriching mitochondria and isolating them from other organelles. The rate at which particles move through the gradient during differential centrifugation is determined by their sedimentation coefficient. This allows researchers to isolate specific cellular organelles, such as mitochondria, by taking advantage of their distinct sedimentation properties.