[Histonet] Immunofluorescence

[koellingr <@t> comcast.net]

Cheryl,
What helped me previously is using a confocal flourescent scope and I'm assuming at LSU there are many.  Looking through a "thick section" with traditional flourescence, you will always be burdened with increased background since you are imaging through the entire section.  With confocal, all that total background becomes irrelavent since , depending on the power of your confocal, you can "optically" section through the thick section, a few micron optical slices at a time.  And 3D image the results to boot by stacking them depending on the software you have.

Raymond Koelling
PhenPath Labs
Seattle, WA

-------------- Original message -------------- 
From: Rene J Buesa <rjbuesa <@t> yahoo.com> 

> No matter what you do, with thick sections you will always get background noise. 
> Try to use thinner sections. 
> Ren$B!&(BJ. 
> 
> Cheryl Crowder wrote: 
> Hello - I have a researcher who has done IHC on her tissues with good 
> success. Now she is trying to do fluorescent antibodies on thick sections 
> to be able to do 3D imaging. I have no experience with this technique. Her 
> problem, tremendous background staining. Can anyone give us any tips for 
> reducing this staining? Thanking you in advance, 
> Cheryl 
> Cheryl Crowder, BA, HTL(ASCP) 
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