When air bubbles is not encouraged in tissue culture? - (Apr/25/2006 )
Hi, does anybody know y we shld try not to introduce air bubbles into the tissue culture flask??? Does air bubbles have any effect on the cells?
Dont air bubbles come out of the media? or are we talking about solid media as agar in microbiology? I dont get it.
When you are dealing with adherant cell lines bubbles in the media can effect cell attachment. When pippetting if you make bubbles in the media it can create aerosols which can lead to contamination in the hood (either cellular or microbial). This can definately affect your cell lines for instance via cross contamination of different lines.
jaimie is 100% right about the aerosols and cell attachment.
bubbles also affect the gas exchange between the media and air, with detrimental consequences for your cells.
I personally can't see why a few bubbles should make any difference in most instances. They disappear fairly quickly. Careful about the containation issue though - if bubbles get into the neck of the flask while subculturing and come into contact with the outside edge of the neck then this could introduce contamination. If maintained properly a hood should be a fairly sterile environment. 99 times out of 100 contamination is introduced because something that should be sterile touches something non-sterile (eg. pipette tip contacting hand or surface of packaging or outside edge of bottle etc). Aerosol contamination should be exceptionally rare if specimens are handled appropriately.
The main problem I've noticed with bubbles is that if they sit across the neck of the flask they can create a film that prevents CO2 exchange (I agree with sudaca). This will lead to your media becoming alkaline and turning pink and the "block" can remain this way for quite some time. Tilting the flask so that medium runs into the neck can cause the same problem, including increasing the risk of contamination.