Re: [Histonet] (kinda) silly questions about IHC


I don't know if this will help, since all I know about is paraffin 
sections, but I mount my tissue on slides from a water bath (which 
will keep bubbles from popping up in between the slide and the 
tissue as long as the water bath isn't boiling or otherwise bubbly 
itself) and leave them to dry overnight on a slide warmer. It's 
never hurt my paraffin sections to let them dry like that, but I 
don't know how that compares to frozen sections.
                                 Rebekah Smith
On Fri Oct 29 09:52:08 EDT 2004, Anna Elisse Beaudin 

> Hello,
>    I have a question about IHC.  I am trying to do multiple 
> labeling on
> sagittal brain sections that were fixed by perfusion.  I cut 
> these
> sections on a cryostat, and I am having a lot of trouble deciding
> whether to do IHC on mounted sections vs. free-floating.  When I 
> try to
> mount fixed sections directly while sectioning, I get air bubbles
> caught between the section and the slide.. the sections just 
> don't seem
> to want to lie smoothly on the slide.  Alternatively, when I 
> stain
> free-floating sections, these long sagittal sections curl up and 
> as
> such I get extremely uneven (and ugly) staining.  My actual 
> (silly)
> question is whether it is possible to collect sections into 
> solution
> (free-floating), mount them, let them dry overnight, and then do 
> IHC on
> them.  I'm torn because I need to let the section dry a little on 
> the
> slide so that they stick, but at the same time I think it might 
> be bad
> for them to dry out?  As you can see, I am very confused, and 
> would
> really appreciate anyone's advice.  Thanks in advance for your 
> help!
> Best,
> Anna Beaudin
> Division of Nutritional Sciences
> Cornell University
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