[Histonet] Vibratome Considerations

[Amos Brooks]

Danielle,
         The thick sections we have been staining have seemed to prefer 
long incubations. Overnight would probably be best. Our PGP on skin 
biopsies at 50 um (free floating sections) have actually had the best 
results when they were left over the weekend but I have a hunch that we 
could do better with a more concentrated antibody. The shaker table is a 
good idea for free floating sections (if that is your plan). If you are 
labeling on a mounted slide the long incubation at room temperature and the 
shaker would probably cause a loss of the reagent.
Amos Brooks


At 12:00 PM 4/12/2004, you wrote:
>Message: 2
>Date: Mon, 12 Apr 2004 09:44:17 -0400
>From: Danielle Zalinski <nsdnz <@t> neuro.hfh.edu>
>Subject: [Histonet] Vibratome Considerations
>To: HistoNet Listserve <histonet <@t> lists.utsouthwestern.edu>
>Message-ID: <407A9D31.70301 <@t> neuro.hfh.edu>
>Content-Type: text/plain
>
>   Hello All,
>Our lab would like to do immunohistochem work in rat brain on vibratome
>sections instead of paraffin.  We are able to get sections about
>50-60um.  Will incubation with primary antibody overnight be sufficient?
>  Is shaking the sections an option?  I was hoping some other labs that
>are working in vibratome sections may have some beginners advice.
>Thanks,
>Danielle Zalinski
>
>Neurosurgery Research
>Henry Ford Health System
>Detroit, MI