 Microbial cryopreservation

Microbial cryopreservation preserves viable microorganisms at very low temperature for later recovery. It provides stored starting material for future experiments, reducing repeated propagation and the associated opportunities for contamination or changes in inherited characteristics. Recovery, purity and identity remain separate properties to verify.^[ref]

Freezing can injure a cell even though low temperature greatly slows its activity. Formation of ice removes liquid water from the surrounding solution and concentrates its dissolved substances; intracellular ice can damage cellular structures. Water movement by osmosis links the external changes to cellular dehydration. Microbial cryoprotectants, including glycerol, help protect the material, but preservation performance depends on the particular organism and conditions.^[ref]

A constructed recovery measurement makes the accounting explicit. Suppose equal original-volume equivalents of a benign yeast suspension give $N_0=1000$ colony-forming units before storage and $N_1=700$ after recovery, using the same assay and correcting for any dilution. The estimated recovery fraction isThese are counts of colony-producing units under the assay conditions. Clumps can form single colonies, so the result does not by itself show that exactly seventy percent of individual cells survived. Nor does a high recovery fraction establish unchanged genotype or every original phenotype.

The point of maintaining a preserved reference is reproducibility: separate experiments can begin from material derived from the same stored stock. Microbial cryopreservation retains frozen material, whereas freeze-drying removes water from frozen material. They are different preservation strategies, whose suitability must be established for the material concerned.^[ref]