[Histonet] question about immunofluorescence staining in mouse spleen


Dear experts,

My boss has asked me about immunofluorescence staining in mouse 
spleen:  How do you get rid of fluorescent background in mouse 
spleen?  I suggested ammonia ethanol quenching which I know works in 
bone marrow.  Is there a better method?  Also, my boss claims that 
the macrophages have autofluorescence (formalin fixed paraffin 
embedded tissue).  I always thought the rbcs would be the 
autofluorescent cells.  Do the macrophages have autofluorescence? 
Finally, is it reasonable to do frozen sections of mouse spleen?  I 
thought frozen sections fixed in ice cold acetone or acetone-methanol 
might solve the problem, but I was told that frozen sections would be 
much to thick to get any good immunofluorescent pictures on.

Thanks for any info and suggestions.

Unknowledgeably yours,

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