How to use the cryopreservation tube scientifically and correctly
The use of the cryopreservation tube is a science, not a simple trilogy such as opening the liquid nitrogen tank, putting it in the cryopreservation tube and closing the liquid nitrogen tank. Scientific and correct use of cryopreservation tubes can avoid the loss of samples and protect the safety of testers.
The use of the cryopreservation tube is a science, not a simple trilogy such as opening the liquid nitrogen tank, putting it in the cryopreservation tube and closing the liquid nitrogen tank. Scientific and correct use of cryopreservation tubes can avoid the loss of samples and protect the safety of testers.
Freezing tube: freezing steps
Wash the cells with preheated PBS solution, suck the solution, and cover the cells with a solution containing trypsin and EDTA (thin liquid layer is enough, and the concentration of trypsin and EDTA needs to be determined according to the cell line).
Incubate cells at 37 ℃ for 3-5 minutes.
After the cells detach from the bottom, the incubation is terminated, the medium containing serum is added, and the cells are gently suspended with a pipette.
Centrifuge the cell suspension (500 x g, 5 min) and resuspension with the medium containing serum.
Cell count.
Centrifuge the cell suspension (500 x g, 5 minutes), remove the supernatant, and resuspension the cells with medium containing serum of appropriate volume.
Mix cells and cryopreservation solution (60% medium, 20% fetal bovine serum, 20% DMSO) in 1:1 volume ratio, and then transfer them to Cryo STM cryopreservation tube. The density of frozen cells is 1-5 × 106 pieces/ml.
Cryo containing cells STM cryopreservation tube is recommended to cool down at a rate of − 1 K/min, and the cryopreservation tube can be placed in a container containing isopropanol at − 70 ℃. If Cryo STM cryopreservation tube stores other samples, which can be directly placed at − 20 ℃, − 70 ℃ or the gas phase of liquid nitrogen. To ensure that the sample is frozen uniformly, 4 mL and 5 mL Cryo The sTM cryopreservation tube needs to be placed in the refrigerator at − 20 ℃ for overnight, and then transferred to the gas phase of − 70 ℃ or liquid nitrogen.
Then transfer Cryo.sTM cryopreservation tube to liquid nitrogen tank. In order to avoid pollution (such as mycoplasma) and safety considerations, please place Cryo.sTM cryopreservation tube in the gas phase of liquid nitrogen, not in the liquid phase.
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Post time: Nov-25-2022