Detecting Fluorescence Lifetimes

Posted by Anke Bielicke on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Detecting-Fluorescence-Lifetimes-tp3333039.html

Hello everybody,

 

I try to measure FRET with a FLIM system from Becker and Hickl containing a MCP detector from Hamamatsu (R3809U-50). This detector is very temperature sensitive (increase of dark counts with increasing temperature). Our room is air conditioned so the room temperature is constant.

 

When I measure CFP over a couple of houres (I measure live cells and allways take a fresh probe) I observe a decrease in lifetime (0.4 ns in 5 hours).

 

How can I avoid this problem??

Has anybody else observed this?

I think it could be the detector and that it is getting hot over the time. Could I cool it down somehow?

 

 

Thanks for any helpful advise.

 

Anke