- Cell fractionation
Cell fractionation is the separation of homogeneous sets, usually
organelles, from a heterogeneous population of cells.
#Disruption (homogenisation) of cells and liberation of organelles.
#Purification of cell components.
Tissue is typically homogenised in an
isotonicbuffer 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 and freeze-thawing.
The solution is homogenised 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.
Cell disruptionfor 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 ceramic filter.
Invariably achieved by
Differential centrifugation- the sequential increase in gravitational force resulting in the sequential separation of organelles according to their density.
* [http://www.microfluidicscorp.com/cell_disruption/index.html Microfluidicscorp - MFIC OTC]
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