Re: [Histonet] Vibratome Considerations

From:Geoff McAuliffe

I have done immunos on vibratome sections of rat brain. If you use a 12 
well tissue culture plate (with its lid) on a shaker table overnight at 
room temp. you should get good results. You will need to add some Triton 
or Tween to the primary antibody (and probably subsequent steps) or do 
something to 'permeabilize' the sections (freeze-thaw or alcohol) so you 
get good penetration of the antibodies. If you have access to a sliding 
microtome with a freezing stage you could obtain sections that way.


Danielle Zalinski wrote:

>  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.
>Danielle Zalinski
>Neurosurgery Research
>Henry Ford Health System
>Detroit, MI
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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