Fwd: Re: [Histonet] mouse brains

[Atoska S. Gentry]

>Date: Thu, 07 Oct 2004 10:26:10 -0500
>To: Gayle Callis <gcallis <@t> montana.edu>
>From: "Atoska S. Gentry" <gentras <@t> vetmed.auburn.edu>
>Subject: Re: [Histonet] mouse brains
>
>
>Gayle, thanks for your reply. Actually, I'm processing these samples for a 
>professor from the Biological Science Department on main campus. And she's 
>conducting various IHC stains for "Tau Filament Formation in Transgenic 
>Mice Expressing P301L Tau" & neurofibrillary changes in CNS. She's working 
>in conjunction with some Spanish Scientists. I believe the samples were 
>shipped from Spain in PFA back in February 04'. When we receive them they 
>are whole brains in PFA  from which coronal slices are taken, processed, 
>and sectioned. The first few sets of brain received a few months ago 
>sectioned much easier, but the two she brought out a couple of weeks ago 
>are requiring a fair amount of trial & error to obtain fairly decent 
>sections. Atoska
>
>
>At 03:11 PM 10/6/2004, you wrote:
>>Atoska,
>>
>>More info please. Are you processing brain slices cut with a matrix 
>>device or whole brain.  We do both and find the time in processing is 
>>critical.   We just did whole mouse brains fixed for three weeks in NBF 
>>for LFB staining and H&E and had wonderful sections but used a longer 
>>processing schedule than for our coronal mouse brain slices.
>>
>>For our coronal sections, we perfuse after anesthetizing animal - first 
>>with saline followed by PLP via heart (a morning protocol then fix for an 
>>additonal 5 hours, slice into 3 mm thick slices with matrix device prior 
>>to a special mouse brain processing schedule on a VIP the same 
>>evening.  PLP has paraformaldehyde so will be similar to your PFA 
>>fixation and are able to obtain excellent coronal sections.  I have not 
>>noticed any differences between longer NBF nor PLP fixation on sectioning 
>>effects.
>>
>>If we fix brain with PFA, we do this overnight at 4C, and if the fixation 
>>needs to be longer, we change the PFA to fresh.   Are you perfusing 
>>(ideal situation) then immersing samples overnight, immersion only, at RT 
>>or 4C.
>>
>>What sectioning problems do you encounter?
>>
>>
>>
>>At 01:08 PM 10/6/2004, you wrote:
>>
>>>Hello, those of you processing & sectioning coronal sections of mouse 
>>>brain; from you experience can prolonged fixation in 4%PFA  be the 
>>>culprit of sectioning problems? What is the maximum fixation time period 
>>>the brain should held in PFA before processing & sectioning? Thanks! Atoska
>>>
>>>
>>>Atoska S. Gentry B.S., HT(ASCP)
>>>Research Assistant III
>>>Scott-Ritchey Research Center
>>>College of Veterinary Medicine
>>>Auburn University, AL  36849
>>>Phone# (334)844-5579  Fax# (334)844-5850
>>>
>>>_______________________________________________
>>>Histonet mailing list
>>>Histonet <@t> lists.utsouthwestern.edu
>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>Gayle Callis
>>MT,HT,HTL(ASCP)
>>Research Histopathology Supervisor
>>Veterinary Molecular Biology
>>Montana State University - Bozeman
>>PO Box 173610
>>Bozeman MT 59717-3610
>>406 994-6367 (lab with voice mail)
>>406 994-4303 (FAX)