Hi,
I am using fluorescent microscope for live cell imaging. The microscope is equiped with incubator that controls, temperature, humidity and CO2 concentration. Unfortunately, the system is not very professional, since it is continously injecting CO2 without any sensor. Yeah, it is adjusted at 5%, but it is open, so after 8h of live imaging the tank of CO2 is completely empty. Deos anyone have a solution for this issue?

Hi,
I dont know what is the configuration of your system.
You meen that the incubator is open for 8H so the temp. is also not 37deg?
If it is just a problem of CO2, did you think to use a HEPES containing medium?
Guy

Hi, for gsovak
I think that you did not get me very well. The sytem is designed for live cell imaging, it controls temperature, humudity and CO2 concentration. the whole "microscope incubator" is closed, it has the same functionality as the "normale incubator" where the cells grow. The only difference is, there is no CO2 sensor.
When you open your " normal incubator" for few seconds, it inject CO2 to establish the 5% concentration and then stop to inject. For the "Microscope incubatore", it injects continouslly CO2 without sensor. it is adjusted to 5% but the flux of CO2 is still open.

Okkkk

I see,
Anyway maybe you can use HEPES containg medium so the pH will remain non acidic.
Guy

Hi

Can you state the make and model of your incubator/microscope. Someone may be able to think up a better solution for you if we know the specs.

In the meantime here are two stop-gap solutions.

1. Invitrogen make a CO2 indenpendent media (cataloge number 18045088). I'm not sure if it is available without phenol red but its worth looking into. Then just turn off your CO2 supply while you are imaging.

2. Can you scavenge an independent CO2 regulator/sensor off an old incubator and plumb it into your inlet pipe?

Please visit this sight for imaging chambers & equipment:

www.mipcompany.com