[Histonet] Sections falling off slides

Thu Nov 11 10:02:13 CST 2004

If you are not drying over night gland do not have time constraints give
that a try!

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317

Disclaimer: 
The information in this e-mail and any of its attachments is confidential
and may contain sensitive information. It should not be used by anyone who
is not the original intended recipient. If you have received this e-mail in
error please inform the sender and delete it from your mailbox or any other
storage devices. National Institute of Allergy and Infectious Diseases shall
not accept liability for any statements made that are sender's own and not
expressly made on behalf of the NIAID by one of its representatives

-----Original Message-----
From: mkundu <@t> purdue.edu [mailto:mkundu <@t> purdue.edu] 
Sent: Thursday, November 11, 2004 6:23 AM
To: Brian Dias; Histology Net List Server (E-Mail)
Cc: Ramos-Vara, Jose A.; Van Alstine, William G.
Subject: Re: [Histonet] Sections falling off slides

Hi ,

I am trying to save the thin section of  biomaterial from falling off in
antigen retival steamer technique.

I am using positive charged superfrost slides and I am doing regular
microtomy with paraffin blocks .
Is there any thing else I can do in addition to prevent from falling?
The sections are very small.

Any suggestions to improve are welcome.

Thank You
MK
----- Original Message -----
From: "Brian Dias" <brian.dias <@t> mail.utexas.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Saturday, November 06, 2004 10:13 PM
Subject: [Histonet] Sections falling off slides

> hi
>   i'm trying to optimize my immunohistochemistry protocol on slides and am

> runnning into a few problems.
> 1. i'd like to know if anyone has any strong opinions (either 
> positive/negative) about conducting such protocols on slides v/s 
> free-floating. i'd like to be able to do this on slides because mounting 
> my sections after performing a free-floating technique is virutally 
> impossible due to the fragile nature and small size of of my sections.
>
> 2. i run into the problem of my sectiions falling off the slide whilst 
> doing my ICC
>
>    the brain is dissected form the skull post-perfusion with saline and 4%

> paraformaldehyde
>    it is then stored in 4% paraformaldehyde for 3 days at 4 C
>    then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days 
> before sectioning on the cryostat at a 30-50um thickness and mounted on 
> Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after 
> they've dried (approx. 20 mins).
>    just before (the night before/2 hours before) performing the ICC, i 
> remove the slides from the freezer and allow to air-dry.
>    immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish
>    1X PBS wash at RT in a dish
>    then normal ICC steps by overlaying solution on slides and here's where

> sections come peeling off.
>
> i'd appreciate any input on the matter
>
> thanks a ton
> regards
> brian
>
>
>
>
> ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ 
> ~ ~ ~ ~ ~ ~ ~ ~
>                                                     Brian George Dias
>
>    Graduate Student (Crews lab)            Office Phone #: 512-475-6738
>    The University of Texas at Austin        E-mail: 
> brian.dias <@t> mail.utexas.edu
>    Institute for Neuroscience                  Website: 
>
<https://webspace.utexas.edu/bgd85/b.htm>https://webspace.utexas.edu/bgd85/b
.htm
>    Patterson 56
>    1 University Station Stop C0930
>    Austin, TX 78712
> ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ 
> ~ ~ ~ ~ ~ ~ ~ ~
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet