Non-mechanical homogenisation procedures

Many cells are susceptible to the appreciable shearing forces that arise on repeated freezing and thawing, or to hypotonic buffers which cause cells to swell up, and in certain cases to lyse this is particularly the case for cells in soft plant and animal tissue. Such treatments only rarely lead to complete cell lysis, the exceptions to this being erythrocytes and reticulocytes which are lysed quantitatively under hypotonic conditions. Non-mechanical homogenisation is of particular relevance to cells like yeast which are refractory to other procedures. One of the simplest procedures for yeast, which can certainly not be described as gentle, is toluene-induced autolysis. This is carried out at room temperature and leads to permeabilisation of the cell walls this causes various hydrolases to be activated causing breakdown not only of the cell structure, but also (undesirably) of many sensitive proteins and nucleic acids in the cell. Consequently, this process is mainly of historical interest. 

The gentlest methods for lysing bacteria and yeasts involve treatment with enzymes. Cell walls are first weakened by the appropriate enzymes, and the sphero-plasts produced are then lysed, either by detergents, osmotic shock, or mechanically. In the case of yeast, whose cell walls are made up from glucans, treatment with 

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