Re: Hypoxia/ pH assay in viable cells

Posted by Farid Jalali on
URL: http://confocal-microscopy-list.275.s1.nabble.com/objective-focal-length-tp1588271p1607264.html

Hello Prabhakar,
For the case of hypoxia, I have used the Oxylite system from Oxford Optronics to measure O2 concentration in vitro during live-cell imaging. The system is quite easy to use and the probes themselves are about $200-$300. They are fibre-optic and quite delicate.
Alternatively, something that we have wanted to try is to use the bioreduced 2-nitroimidizoles EF5 or pimonidazole and stain indirectly using immunofluorescence (there is a Cy3-conjugated ab available to EF5). I know its not real time, but you may be able to develop a calibration curve for future experiments. See any publication by CJ Koch for the use of EF5.
Best
Farid

On Tue, Dec 2, 2008 at 4:25 PM, B. Prabhakar Pandian <[hidden email]> wrote:

Hello,
         Can someone tell me what fluroescent asssays are currently available for studying pH and hypoxia changes in a cell culture real-time.

Thanks,

-Prabhakar