IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

immunofluor.-photobleaching

Robin L. Wright wrightr at ZOOLOGY.WASHINGTON.EDU
Thu Aug 25 11:38:27 EST 1994


Are you mounting your coverslips on your immunfluorescence slides  
with a bleach retardant solution?  We have been using Citifluor for  
several years and it works very well.  But, there are numerous other  
alternatives.  Also, it sounds like your fluorescence light source  
needs to be adjusted to give you a uniform signal.  Opitimally, the  
entire field should be uniformly bright, so that you don't have dark  
edges.  


>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
First, I'd like to thank everyone for the advice on getting my yeast  
cells to 

stick to the slide. I've had some success. The problem now is  
bleaching. My
cells stain, I can see them in the periphery of the field of view,  
but as 

soon as I bring them into the center for a good view, they VERY  
RAPIDLY 

bleach. My secondary ab is DTAF conjugated (same excitation &  
emission 

wavelengths as FITC). Is this bleaching a result of unstable  
secondary? my 

staining technique? or the UV source? Any references or info would be  
greatly 

appreciated.  


Thanks in advance!



More information about the Yeast mailing list

Send comments to us at biosci-help [At] net.bio.net