[Histonet] Any fixative to reduce background in immunofluorescence?

[ajennings <@t> unmc.edu]

Clarification please
are you using a secondary that is excited between 470-490? FITC?
if so........does your background fluorescence appear to be more yellow
than green?
"paraformaldehyde" is not going to fix this particular problem
is it confluent over the entire tissue? found in the blood cells?

it may be as simple of a fix as changing your tag to something that excites
530-550

anita








                                                                           
             "Tan, MinHan"                                                 
             <MinHan.Tan <@t> vai.o                                             
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                                       [Histonet] Any fixative to reduce   
                                       background in immunofluorescence?   
             10/27/2004 12:14                                              
             PM                                                            
                                                                           
                                                                           
                                                                           
                                                                           




Hi,

I am preparing to do some immunofluorescence staining in kidney tissue
with cd31.

Some preliminary work in formalin fixed paraffin embedded tissue has
yielded quite significant background fluorescence.

A colleague suggested that I use frozen tissue, but the morphology is
quite unacceptable.

Another colleague suggested paraformaldehyde, but that is really
inconvenient (preparing this every morning!?)

Any advice on a good commercial fixative for this purpose, that doesn't
destroy vessel antigens?

Thanks.

Min-Han

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