LN2 is getting into some of the vials during preservation in liquid nitrogen. I use internal threaded cryovials with 'O' ring (Corning/ Nunc). Can someone suggest a better brand of cryovials?
Hello Srinivas,
You could try with the Nalgene cryogenic tubes Cat. 5000-0020. These ones has external threads so the aseptic technique is enhanced and the sealing is generally tight enough. It worked ok in our lab.
The external thread lids which screw into the vial are annoying with respect to sterility but I was told that this configuration was better for preventing leakage of liquid nitrogen into the tube. Definitely contact Corning and Nalgene sales reps and find out what they think on this topic. Both companies make good cryotubes.
Dear laura
We use 0.5-ml IMV French straws for packing and freezing of semen samples in our laboratory and it works OK.
I actually ONLY use Corning, specifically because of leakage problems I've had with other manufacturers.
I use product number 430659, "cryogenic vial, self standing, silicone washer, external thread, marking spot, non pyrogenic, polyproylene, sterile"
It's a 2.0ml vial, but I only ever put 1ml of fluid inside it, because as the slurry expands during freezing, it can swell the tube, which can cause cracking, and leaks later.
I second Jeffrey's suggestion to call the manufacturer and tell them about your troubles, it may be a bad lot of tubes. If other customers have complained, they'll be able to tell you that, and suggest an alternative product (which they should give you for free).
The plastic cryovials are not meant to be used submerged in liquid nitrogen. If you read carefully the manufacturers instruction you will see they are to be used for storage in the vapor phase of liquid nitrogen. Almost all the cryovials mentioned in this discussion will allow some liquid nitrogen inside if stored submerged. That becomes very dangerous when you retrieve such vial out of the liquid nitrogen. The liquid inside the vial start boiling very fast and the pressure inside the vial increases so quickly that the vial might explode. I have seen that happen. Note that 1cc liquid nitrogen produces some 694cc vapor. We store millions of cells in vapor phase nitrogen for years without any problem. Temperature of vapor phase storage is -150 degrees C while liquid is -196 degrees C. If you want to store in liquid phase nitrogen you might want to consider having the cells in glass ampules hermetically closed.
Dear Sir,
Please go for TPP brand Cryo tubes (O-ring free technology!), safe for both vapor and snap freezing protocols!
www.tpp-india.com & www.tpp.ch
Cook Regentec designed CellSeal closed-system cryogenic vials expressly for the purpose of liquid nitrogen preservation of cell samples. The vials are available in 2 mL and 5 mL sizes, and form part of a portfolio of products intended to standardize your entire freeze-store-thaw-deliver procedures for cell therapy products. They are suitable for both manual and automated filling, and are currently being used in multiple cell therapy clinical trials world wide.
[Disclosure: I am an independent consultant, but am currently affiliated with Cook Regentec.]
the ones with the rubber kgasket. corning 430659 wont leak or explode if you seal double tight.
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