Vials stored in liquid nitrogen - risk of explosion

A recent accident in one of our laboratories should serve as a reminder to us all of the possibility that
vials stored in the liquid phase of liquid nitrogen storage systems can explode after being removed
from the storage vessel. It is preferable to store vials in the vapour phase rather than the liquid phase
so as to eliminate the possibility of explosions.

What happened?
The accident occurred after the vial had been removed from storage in the liquid phase of a liquid
nitrogen storage vessel. The vial was one designed and manufactured for storage at cryogenic
temperatures. It exploded and part of the vial became embedded in the laboratory worker's hand.
Fortunately the injury was slight but the potential for more serious injury if the circumstances had
been slightly different was evident. Severe injuries from exploding vials have been recorded at a
number of other research institutions.

Why did it happen?

When a sealed vial is cooled to the temperature inside a liquid nitrogen storage vessel, the pressure
inside the vial will decrease resulting in a significant pressure difference between the inside and
outside of the vial. If the cap on the vial performs as intended this will not be a problem. The vial will
remain sealed and the pressure inside the vial will remain at the reduced level while the vial is in
storage. If however the seal between cap and vial were to leak slightly and the vial were to be placed
in the liquid phase inside a liquid nitrogen storage vessel, liquid nitrogen would be drawn into the vial.
When the vial was then removed from storage and started to warm up, the liquid nitrogen inside the
tube would rapidly turn to gas which would occupy about 700 times the liquid volume at atmospheric
pressure. Because the gas was contained inside the vial there would be a rapid build up of pressure
leading to an explosion. (Because the cap leaks only slightly it is unable to vent the rapid build up of
pressure.) It is thought that this is what happened to cause the accident in our laboratory.

What do the manufacturers of vials say about this?

Manufacturers of vials used for liquid nitrogen storage recognise that this can happen and that they
might be held liable for any injuries which result. Because of this most manufacturers now state that
their vials should be used in vapour phase cryogenic storage and not be immersed in the liquid
phase.

What should we do now?

It is recommended that vials be stored only in the vapour phase in liquid nitrogen storage vessels and
not in the liquid phase. The vials should however still be of a type designed and constructed for the
temperatures which will be found in a liquid nitrogen storage system.

What should we do if we wish to continue to store vials in the liquid phase of liquid nitrogen
storage systems?
If research groups wish to continue to store vials in the liquid phase great care should be taken when
they are removed from the storage vessel. It can never be guaranteed that a vial has not leaked
causing liquid nitrogen to enter the vial. Every vial should be treated as if it could explode and
cryoprotective gloves, full face-shield and a fastened laboratory coat should always be worn while
handling the vials. In addition forceps should be used to handle the vials where possible. Many
institutions recommend placing the vial inside some form of secondary containment (e.g. sandwich
box + lid) immediately after removing from liquid nitrogen to allow it to thaw; if the vial is going to
explode, it is likely to do so very soon after removal. Finally consideration should be given to sealing
the individual vials in cryoflex tubing or equivalent which is designed to prevent penetration by liquid
nitrogen.

Where can we get further information?

Sources of information include the Cryopreservation Guide produced by Thermo Scientific:
http://www.fisher.co.uk/techzone/pdfs/cryopreservation_manual_2010.pdf (see the section on safety
considerations). You can also contact the University Safety Adviser.

Nigel Corby
University Safety Adviser
January 2011