CO2 gassing using Zeiss spinning disc confocal

Posted by Julia Edgar on
URL: http://confocal-microscopy-list.275.s1.nabble.com/CO2-gassing-using-Zeiss-spinning-disc-confocal-tp7586692.html

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Dear All
I’m using a Zeiss spinning disc confocal microscope with small heated incubation stage and ‘gassing’ lid.
The problem is that the pH of the media changes over time and becomes rather alkaline (turns blue pink). Motor proteins don’t behave normally when pH is not physiological, so this is potentially a big problem.

Do you have any tips to ensure the 5% CO2 reaches the imaging dish (a 35 mm Petri-dish) that sits between the incubation insert and the gassing lid?
The tubing connecting the gassing lid to the water bottle (into which the CO2 is pumped) is rather long. Would it help to shorten it?
Does the volume of water in the water bottle matter?
How can I ensure the little inlet pipes in the gassing lid are not blocked?

Many thanks
Julia
University of Glasgow.