- Cell disruption by nitrogen decompression
Cell disruption by rapid
decompressionis one of several methods of cell disruptionand is also called explosive decompressionor cell bomb.
The technique is used to:
Homogenizecells and tissues
* Release intact
* Release labile
* Produce uniform and repeatable
homogenates without subjecting the sample to extreme chemical or physical stress.
According to manufacturers of nitrogen decompression devices, the method is particularly well suited for treating
mammalianand other membrane bound cells. It has also been used successfully for treating plant cells, for releasing virus from fertilized eggs and for treating fragile bacteria. It is not recommended for untreated bacterial cells. Yeast, fungus, sporesand other materials with tough cell walls do not respond well to this method.
How it works
Large quantities of nitrogen are first dissolved in the cell under high pressure within a suitable
pressure vessel. Then, when the gas pressure is suddenly released, the nitrogen comes out of the solution as expanding bubbles that stretch the membranes of each cell until they rupture and release the contents of the cell.
Nitrogen decompression is claimed to be more protective of
enzymes and organelles than ultrasonicand mechanical homogenizing methods and to compare favorably to the controlled disruptive action obtained in a PTFE and glass mortar and pestle homogenizer. While other disruptive methods depend upon friction or a mechanical shearing action that generate heat, the nitrogen decompression procedure is accompanied by an adiabatic expansion that cools the sample instead of heating it.
The blanket of inert nitrogen gas that saturates the cell suspension and the homogenate offers protection against
oxidationof cell components. Although other gases: carbon dioxide, nitrous oxide, carbon monoxideand compressed air have been used in this technique, nitrogenis preferred because of its non-reactive nature and because it does not alter the pH of the suspending medium. In addition, nitrogen is preferred because it is generally available at low cost and at pressures suitable for this procedure. Once released, subcellularsubstances are not exposed to continued attritionthat might denature the sample or produce unwanted damage. There is no need to watch for a peak between enzyme activity and percent disruption. Since nitrogen bubbles are generated within each cell, the same disruptive force is applied uniformly throughout the sample, thus ensuring unusual uniformity in the product. Cell-free homogenates can be produced.
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