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Exploding cryovials - tips to prevent? - (Jan/19/2008 )

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Hi,

My PI was recently thawing out some cells from LN2 storage. The cells were stored in the lowest rack of the LN2 tank, so they were in liquid. Anyway, during thawing in the water bath within a few minutes the vial exploded, losing the sample. I've read a few posts on this forum about exploding vials already. I was wondering if you can offer any tips to prevent this while still storing the cells in LN2. My PI mentioned we should open the cap slightly...but...I'm afraid of contamination. Do you just want to open it so the rubber is not sealing it anymore?

Thanks,
- Eli

-eli2k-

QUOTE (eli2k @ Jan 19 2008, 12:55 PM)
Hi,

My PI was recently thawing out some cells from LN2 storage. The cells were stored in the lowest rack of the LN2 tank, so they were in liquid. Anyway, during thawing in the water bath within a few minutes the vial exploded, losing the sample. I've read a few posts on this forum about exploding vials already. I was wondering if you can offer any tips to prevent this while still storing the cells in LN2. My PI mentioned we should open the cap slightly...but...I'm afraid of contamination. Do you just want to open it so the rubber is not sealing it anymore?

Thanks,
- Eli


thawing of frozen eukaryotes should be performed slowly, f.i. starting thawing on ice;

good quality vials should not explode even with fast thawing...

-The Bearer-

I think the key to this problem is when you freeze the vials, don't screw tight! Only use vials that are intended for freezing cells. I have never experienced such explosion unless I use eppendorf tubes.

-pcrman-

The problem is that these cells were purchased from a company, not ones that I froze down myself.

-eli2k-

Once you remove the vials from the liquid nitrogen tank, if they are tight screwed, loose them a little bit. When thawing them in 37C water bath, make sure the water doesn't get into the vials.

-pcrman-

QUOTE (pcrman @ Jan 19 2008, 05:26 PM)
Once you remove the vials from the liquid nitrogen tank, if they are tight screwed, loose them a little bit. When thawing them in 37C water bath, make sure the water doesn't get into the vials.


Yea, I wanted to ask, how much is a little bit? One complete turn open, assuming it takes around 3 complete turns to completely open?

-eli2k-

QUOTE (eli2k @ Jan 19 2008, 12:55 PM)
Hi,

My PI was recently thawing out some cells from LN2 storage. The cells were stored in the lowest rack of the LN2 tank, so they were in liquid. Anyway, during thawing in the water bath within a few minutes the vial exploded, losing the sample. I've read a few posts on this forum about exploding vials already. I was wondering if you can offer any tips to prevent this while still storing the cells in LN2. My PI mentioned we should open the cap slightly...but...I'm afraid of contamination. Do you just want to open it so the rubber is not sealing it anymore?

Thanks,
- Eli



Dear Eli,

There are many types of cryovials. The ones that reduce this happening is the ones that have and INTERNAL THREAD and O-RING. We have been using this type for many years and have very very rarely had problems. Vials can be stored in VAPOUR PHASE instead of liquid phase...this further reduces the probability of exploding vials.
Always wear PPE when handling liquid nitrogen and ALWAYS assume that there is RISK involved.

Kindest regards.

Rhombus.

-Rhombus-

QUOTE (The Bearer @ Jan 19 2008, 08:52 PM)
thawing of frozen eukaryotes should be performed slowly, f.i. starting thawing on ice;


I thought it was "freeze slow / thaw fast"

if it is then you can do it in your hand (gloved of course)

i use the same ones as rhombus (alphalabs to be specific) and dont have that problem

dom

-Dominic-

QUOTE (Dominic @ Jan 21 2008, 09:41 AM)
QUOTE (The Bearer @ Jan 19 2008, 08:52 PM)
thawing of frozen eukaryotes should be performed slowly, f.i. starting thawing on ice;


I thought it was "freeze slow / thaw fast"

if it is then you can do it in your hand (gloved of course)

i use the same ones as rhombus (alphalabs to be specific) and dont have that problem

dom


Dom,

Exactly my method....your not one of my old students are you ?

Regards

Rhombus

-Rhombus-

QUOTE (eli2k @ Jan 19 2008, 04:22 PM)
The problem is that these cells were purchased from a company, not ones that I froze down myself.


All the vials I've ever gotten from ATCC have said to store in the Vapor Phase, which would keep any LN2 from getting into the vial and causing it to explode upon thawing. I've not had problems with thawing ATCC vials after storing them in the liquid phase, but your mileage may vary with other vendors.

TheSquire

-TheSquire-

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