How can I isolate mitochondria from cells?

The process to isolate mitochondria from cells involves two steps. The first step is cell disruption, which involves breaking open of the cell to release its cellular structure. It is generally referred to as cell lysis. There are different methods available to perform cell lysis. Physical techniques, like osmotic shock, simple freeze and grind, etc., can be used to physically break open the cell. Chemical methods like using detergents and biological lysis methods like using enzymes are also available to degrade the cell walls.

The second step is centrifugation. Mitochondria and other components of the broken cell can be separated by centrifugal force. All the broken cell components are rotated at different speeds. Depending on the size and density of the cell component, each component experiences different centrifugal force and deposits at the bottom. At a relatively high speed, mitochondria can be isolated form other cell components. A pellet of mitochondria will be deposited. Multiple rounds of centrifugation can be applied to get pure mitochondria.