[Histonet] [Fwd: Timm's staining human tissue]

From:Mike King

-------- Original Message --------
Subject: Timm's staining human tissue
Date: Thu, 04 Nov 2004 15:26:35 -0500
From: Mike King 
To: histonet-request@lists.utsouthwestern.edu

As usual Dr. Kiernan is spot on in explaining why Timm's won't work in
FFPE specimens.  It is possible to do obtain acceptable Timm's
histochemistry after immersion of fresh biopsy material in sulfide
solutions, followed by immersion fixation.  See our paper
Murray KD, Isackson PJ, Eskin TA, King MA, Montesinos SP, Abraham LA,
Roper SN.  Altered mRNA expression for brain-derived neurotrophic factor
and type II calcium/calmodulin-dependent protein kinase in the
hippocampus of patients with intractable temporal lobe epilepsy.  J Comp
Neurol. 2000 Mar 20;418(4):411-22.

I'll attach a pdf directly to you.  The technique is likely to work foa
autopsy tissue too.

original post:
From: Chris.Goodall@bristol.ac.uk
Subject: [Histonet] Timm silver sulphide method for light microscopic
	localisation of heavy metals.
To: histonet@lists.utsouthwestern.edu
Message-ID: <1099576092.418a331c8b602@webmail.bris.ac.uk>
Content-Type: text/plain

Dear All,
     I have been asked to try the Timm method on some FFPE samples of
human brain. The papers I have read give methods for perfusing rat
brain with sodium sulphide solution followed by 3% gluteraldehyde in
0.15M phosphate buffer, and further treatment with sodium sulphide
solution, or, post mortem brain samples snap frozen and frozen
sectioned. However health and safety rules here do not permit frozen
sectioning of unfixed human brain, or the use of gluteradehyde in the
embalming room, so I am going to attempt this method on formalin fixed
samples.My question is, has anyone experience of this method or any
suggestions? It is mentioned that autopsy material left in situ for one
or two days post mortem with no fixation may have enough endogenous
sulphide ions or sulphide groups in the tissue due to autolytic
activity and sulphide treatment may not be necessary, or if it is
necessary, does anyone have experience of timing in the sulphide
solution and is post fixation necessary after the initial formalin
fixation. The paper also mentions that to prevent loss of
metallosulphides present in the tissue the temperature of the wax
should not exceed 50C which means the use of the dreaded low
temperature wax, has anyone used conventional paraffin wax and got away
with it?
Sorry for the long request,and thank you,
Chris Goodall


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