METHOD OF CRYOPRESERVATION OF MARINE MICROALGAE Russian patent published in 2013 - IPC A01N1/02 

FIELD: agriculture.

SUBSTANCE: invention relates to the field of cryobiology, cell biology, marine biotechnology and hydrobiology. Processing of marine microalgae cells is carried out with cryoprotective mixture containing penetrating and non-penetrating cryoprotectant and a nutrient medium. Freezing is carried out, followed by preservation in liquid nitrogen. Sterile seawater is taken as the nutrient medium in the following ratio of components, wt %: penetrating cryoprotectant 5-7.5%, non-penetrating cryoprotectant 1.5-3%, sterile sea water - the rest. Prior to freezing, the cells of marine microalgae are incubated in the ice bath for at least 10 minutes. Freezing is carried out in three phases: first, cooled to -25°C at 0.9-1.1 °C/min.; then cooled to -75°C at 2-2.5 °C/min.; then cooled to -196°C, placing marine microalgae cells in liquid nitrogen. Dimethyl sulfoxide (DMSO) or ethylene glycol can be used as penetrating cryoprotectant. Trehalose or polyvinylpyrrolidone (PVP) can be used as non-penetrating cryoprotectant.

EFFECT: invention enables to increase the yield of viable, functionally active cells of microalgae after freezing-thawing and to restore their culture for a week after cryopreservation, while providing a quantitative assessment of the functional state of the microalgae.

3 cl, 1 dwg, 3 tbl, 5 ex