Greiner cryotube safety instructions
Improper use of cryotubes, explosion, biosecurity threats and personal injury risks, improper use of frozen tubes, explosions, biosecurity threats and personal injury risks, improper use of frozen tubes, explosions, biosecurity threats and Risk of personal injury
Cell cryopreservation process cell cryopreservation process
Wash the cells with pre-warmed PBS, discard the cells, discard the PBS, add the mixture, add the mixture, and add the trypsin digest solution containing EDTAEDTAEDTAEDTA to remove the cells. Concentrate the chemical solution to eliminate the cells (thinly cover the surface, the concentration of the chemical solution to eliminate the cells (thin cover the surface, the concentration must be adjusted according to different cell lines). Adjust according to different cell lines).
Digest cells at 37 °C for 3–5 minutes without overdigestion.
Once the cells are detached from the bottom, the digestion can be stopped with the serum-containing medium and the pipette can be lightly blown. Once the cells are detached from the bottom, the digestion can be stopped with the serum-containing medium and the pipette can be lightly blown. Once the cells are detached from the bottom, the serum can be used to terminate the digestion. And the pipette is lightly blown. Once the cells are detached from the bottom, the digestion can be stopped with the serum-containing medium and the pipette can be lightly blown. Once the cells are detached from the bottom, the cells can be stopped by the serum-containing medium and the pipette can be lightly blown to resuspend the cells.
Centrifuge (500 00 00 × g, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 min) cell suspension to precipitate,) cell resuspension to precipitate, discard the supernatant The cell pellet was resuspended in serum-containing medium.
The cells were counted (Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber Neubauer chamber).
Centrifuge (500 500 500 500 × g, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 ming, 5 min) the cell suspension and discard the supernatant. The cells were resuspended in an appropriate volume of blood-containing medium and the supernatant was discarded. Resuspend the cells in an appropriate volume of blood containing medium.
Resuspend the cell suspension in a 1:11:1 ratio to the cryopreservation solution (60% medium, medium, 20% FCS20% FCS20% FCS 20% FCS 20% FCS, 20% DMSO 20% DMSO20) % DMSO 20% DMSO 20% DMSO 20% DMSO 20% DMSO 20% DMSO) After mixing, transfer to Cryo.sCryo.sCryo.s Cryo.sCryo.sTM cryotubes. The concentration of cells should be 1–5 × 10 6 cells/ml.
The cryotube containing cells shall be frozen at a cooling rate of .1 K/min1 K/min1 K/min1 K/min1 K/min1 K/min1 K/min. The cooling rate at which the tube is inserted is frozen. The cooling rate at which the tube is inserted is frozen. The above requirements can be achieved by inserting the tube in a freezer compartment containing isopropyl alcohol and placing it in a refrigerator at .70 °C. If the refrigerator can achieve the above requirements. If the refrigerator can achieve the above requirements. If other types of samples are required in the cryopreservation solution, Cryo.sCryo.sCryo.s Cryo.sCryo.sTM cryotubes should be stored at .20 °C. The vapor phase layer at 70 ° C or liquid nitrogen is directly frozen. To ensure uniformity of each sample, 4 ml and 5 ml Cryo.sCryo.sCryo.s Cryo.sCryo.sTM cryotubes should be stored overnight at .20 °C and then transferred to 70 °C or liquid nitrogen gas phase layer. .
The Cryo.s Cryo.s Cryo.s Cryo.s Cryo.sTM cryotube was then transferred to a liquid nitrogen tank. To avoid contamination (such as mycoplasma) and transfer the frozen tube to the liquid nitrogen tank. To avoid contamination (such as mycoplasma) and transfer the frozen tube to the liquid nitrogen tank. To avoid contamination (such as mycoplasma) and transfer the frozen tube to the liquid nitrogen tank. In order to avoid contamination (such as mycoplasma) and to take into account the requirements of safety precautions, it is recommended that the Cryo.sCryo.sCryo.s Cryo.sCryo.sTM cryotube be placed in the gas phase above the liquid nitrogen, taking into account the safety precautions. Layer instead of liquid nitrogen.
Cell recovery process
Immediately after removing the cryotube from the liquid nitrogen tank, it was thawed in a 37 ° C water bath. The thawing time was about 1–2 min. When thawing, it was necessary to shake the cryotube to melt it as soon as possible. The faster the thawing process in this step, the better.
Transfer the thawed cell suspension to a 15 ml centrifuge tube and immediately add a quantity of serum-containing medium for mixing.
The cells were resuspended by centrifugation (500 × g, 5 min) and the supernatant was discarded. The cell pellet is resuspended in an appropriate volume of serum-containing medium and dispensed into one or more cell culture flasks.
Please follow the recommended concentration at the time of cell inoculation.
The cells need to be statically cultured for the next 12 hours.
Cell exchange can be performed after 24–48 h.
Suggestions for safe use of Cryo.sTM cryotubes
When using a cryotube to store samples, it is strictly required to store the cryotubes in a vapor phase layer of liquid nitrogen or in a refrigerator. If the cryotubes are stored in a liquid nitrogen liquid, liquid nitrogen has a certain probability of seeping into the inside of the cryotubes. Nitrogenation of the liquid during resuscitation leads to unbalanced pressure inside and outside the tube, which is highly prone to bursting of the cryotube and is biohazardous.
Operate the cryotubes for resuscitation and use safety equipment throughout the entire process. It is recommended to wear lab coats, cotton gloves and operate on a safe bench. Wear goggles or a face shield if possible. Be extra careful because the indoor temperature in summer will be higher than in winter.
During the storage of cryopreserved cells, the freezing temperature of the cryotubes must be uniform. Uneven freezing will result in the formation of ice plugs that will inhibit the temperature transfer of liquids on both sides and create dangerously high pressures and damage to the cryotubes.
The amount of frozen sample should not exceed the maximum working volume required by the cryotube.
Beijing Unicorn Biotechnology Co., Ltd.
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