RE: [Histonet] BRDU free floating vibratome sections

From:"Charles Scouten"

 The Histodipper from the Vibratome Company is designed for free floating sections cut on the Vibratome(TM).  It also conserves reagents.  See the following link:

Charles W.  Scouten, Ph.D. 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 

-----Original Message-----
From: [] On Behalf Of Sarah Jones
Sent: Thursday, August 05, 2004 4:57 PM
Subject: Re: [Histonet] BRDU free floating vibratome sections

Hi Kimberle,
   I've done quite a bit of vibratome sectioning and free floating staining.  It's quite challenging!  Two ways I can think of that might help you.  One would be to use the Shandon coverplate system.  The sections would be floated onto slides and then the coverplate carefully placed on to the slide.  It then fits into a holder and it gravity feeds the reagents over the slide.  It uses a very small amount of reagent. 
The other way is to handle the free floating sections in netwells or you can make your own using open cassettes and small petri dishes.  The most important steps would be making sure the rinsing steps were sufficient. 
The netwell/petri dish method would use a much greater amount of reagent.  Hope this helps.  Sarah

Sarah Jones, HT(ASCP)
Histology Lab
Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499

>>> "Kimberle M. Jacobs"  08/05/04 3:32 PM >>>
Hi I have searched the archives and found a lot of discussion of BRDU, but no answer to the question if there is a protocol out there for formaldehyde-fixed, vibratome-cut, free floating sections.  Actually we could mount them on slides as well, I just don't want to do the whole paraffin embedding thing.
Also is it really necessary to have thin sections if you are going to use pepsin and acid, etc. ??

We could probably cut as thin as 20 um, but would prefer to use 40 or 60 um.

We will be doing this in pregnant ferrets (E36, gestation is ~42 days) and examining the brains at ages P0-P15.

Thanks very much for any help!!

Kimberle M. Jacobs, Ph.D.
Assistant Professor
Department of Anatomy and Neurobiology
Virginia Commonwealth University
P.O. Box 980709
Richmond, VA  23298-0709
(804) 827-2135
FAX: (804) 828-9477

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