We never had any problem with the observation temperature, but GFP is extremely sensitive to the temperature at which folding occurs. The latter is widely divergent depending also on the mutant used (some are worse than the wild type). Thus, if you have a chance of keeping the cells you want at room temperature for a few hours, this may critically help. you might also observe a higher GFP fluorescence in cells from the tail vs bone marrow (the tail is at lower temperature). Saverio Alberti Head, Lab. of Experimental Oncology Department of Cell Biology and Oncology Consorzio Mario Negri Sud 66030 Santa Maria Imbaro (Chieti), Italy Phone: (39) 872-570.293 FAX: (39) 872-570.412 E-mail: alberti@cmns.mnegri.it On Wed, 7 Jul 1999, Geoffrey Osborne wrote: > > Hi All, > I looked around but can't find any references to.... > Whether the level fluorescence of GFP is related to the temperature at > which it is exposed to an appropriate light source? > My query is related to the lack of detectable GFP signal by flow cytometry, > from a GFP + transgenic mouse (the skin on the tail glows for example) bone > marrow cells. If the temperature is important, then in the mammalian system > the GFP may not be suitbale for the researchers needs. > > Thanks in advance, > Geoff > ====================================================================== > Geoffrey Osborne | ____ __ o Ahh! > Flow Cytometry (FACS LAB) | __ `\ <,_ > John Curtin School of Medical Research, | __ (*)/ (*) > Australian National University, | ==============| > CANBERRA, AUSTRALIA. | |--| > Email: Geoff.Osborne@anu.edu.au | |--|... > Phone: 61 2 6249 3694 | > FAX: 61 2 6249 2595 | > -----Surfing the Web?: Try http://jcsmr.anu.edu.au/facshome.html > ====================================================================== >
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