METHOD FOR CRYOPRESERVATION OF BIOLOGICAL SAMPLES UNDER PRESSURE AND DEVICE FOR ITS IMPLEMENTATION Russian patent published in 2019 - IPC A01N1/02 F25D31/00 

FIELD: cryogenics.

SUBSTANCE: invention relates to cryopreservation for long-term storage of biological samples. Method of cryopreservation of biological sample includes saturation with cryoprotector solution, placing sample in inner volume of high pressure chamber, sample vitrification in the chamber with increase of the hydraulic pressure in the chamber volume to level of 500–2,100 atm. depending on type of biological sample. Further, the verified sample is transferred for storage. At the first stage of the sample cooling, pressure is increased at a rate of not more than 50 atm/s, pressure build-up is started at temperature in range from 0 °C to -50 °C and high pressure formation is completed in the temperature range from -35 to -100 °C. Second step of cooling the sample to the glass transition temperature is carried out at high pressure in the isobaric mode. When the temperature of the biological sample reaches 1–10 °C below the vitrification temperature proceeds to the third stage of cooling in the isochoric mode until the sample storage temperature is reached. After storage of the biological sample in the chamber, the high pressure is restored and the first stage of heating the biological sample from the storage temperature to the glass transition temperature in the isochoric mode is carried out. At second stage of heating from glass transition temperature to temperature range from -100 to -35 ° heating is carried out in isobaric mode, maintaining formed high pressure on sample. At third stage of heating in temperature range from -50 to 0 °C is gradually reduced pressure to normobaric state at a rate of not more than 50 atm/s.

EFFECT: disclosed method of cryopreservation of a biological sample enables to increase viability of a biological sample by eliminating cracking of a sample during vitrification.

18 cl, 3 dwg, 2 tbl