Cell fractionation is the separation of homogeneous sets, usually organelles, from a heterogeneous population of cells.
There are three principal steps involved:
Tissue is typically homogenized in an isotonic buffer solution using a variety of mechanisms. A 'Potter-Elvehjem homogeniser' is often used as it is relatively gentle. Other procedures include grinding, mincing, chopping, pressure changes, osmotic shock, freeze-thawing, and ultra-sound h...
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Cell fractionation is the separation of homogeneous sets, usually organelles, from a heterogeneous population of cells.
There are three principal steps involved:
Tissue is typically homogenized in an isotonic buffer solution using a variety of mechanisms. A 'Potter-Elvehjem homogeniser' is often used as it is relatively gentle. Other procedures include grinding, mincing, chopping, pressure changes, osmotic shock, freeze-thawing, and ultra-sound homogenization.
The solution is homogenized in an isotonic solution to stop osmotic damage, with a pH buffer to regulate pH, and at an ice-cold temperature to prevent enzyme damage. The organelles are kept either cold, isotonic or buffered.
See Cell disruption for further details.
This step may not be necessary depending on the source of the cells. Animal tissue however is likely to yield connective tissue which must be removed. Commonly, filtration is achieved either by pouring through gauze or with a suction filter and the relevant grade...
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