Re: Re: [Histonet] Cryostat sections mounting with 0.1M PB solution onto slides

[TF]

How about to use a drop of water?

Sometimes the section is a bit big, and it is hard to mount the section very flat on slides without a bubble...

2009-04-20 



TF 



发件人： Paula Pierce 
发送时间： 2009-04-03  23:59:43 
收件人： tifei 
抄送： 
主题： Re: [Histonet] Cryostat sections mounting with 0.1M PB solution onto slides 
 
Hi TF,

you should not place the drop of buffer on the slide under the section. The section will wash off. You will get buffer crystals under the section during drying and although they will dissolve out later, they will cause the section to not stick to the slide where they are.

Place the section directly on a warm (room temperature) slide and the section will adhere.

Paula





From: TF <tifei <@t> foxmail.com>
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, April 3, 2009 10:43:03 AM
Subject: [Histonet] Cryostat sections mounting with 0.1M PB solution onto slides

After we cut the brain sections on a cryostat (OCT embedded), we brush a drop of 0.1M PB on to the slide before mounting the sections.
Do anyone know the side effect of this - for example, the sections will peel off during immunostaining?

Another question is about the dry time after mounting and before staining.
I know some people asked similar questions before, but they are using fresh tissue for frozen sections.
We here perfuse the animal, post-fix the tissue and sink it in sucrose before making cryostat sections.
So we may dry the sections in air up to weeks.
I would like to ask about the proper heating time (with a heat plate) before dry the sections up in a fumehood.
Should we heat the sections at 60 degree for 10 min or 2 hours shortly after mounting?

Thanks very much.


2009-04-03 



TF 
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