[Histonet] Re: Timms stain in fresh frozen cryosections

From:Delatour =?iso-8859-1?Q?Beno=EEt?=

Hi Anna,
You can simply incubate mounted slices in a 1.2% NaS.9H2O solution (in 
buffer w/v : 0.6 g / 50 cc of PB 0.1M) for 30 min. and then, after 
rinsings, proceed with TIMM staining.
It works well with the limit that cutting fresh unfixed brains and doing 
repeated incubations and baths is risky (tissue goes away!). To minimize 
this, cut thin (<10µm) and slowly. Let the sections dry.

Other alternatives
1) mice are alive : perfuse them with Na2S followed by fixative ; gives 
excellent results.
2) brains have not yet been cut : the simplest method for lazy people is to=20
do "immersion autometallography" by incubating tissue in a solution of 
Na2S, then fixation in formadehyde. Once slices are obtained, proceed to 
TIMM staining.


--------"If you think research is expensive, try disease"--------

B. Delatour
Laboratoire de Neurobiologie de l'Apprentissage,
de la Mémoire & de la Communication,
NAMC, CNRS UMR 8620, Bât 446
Université Paris-Sud
91405 Orsay Cedex, FRANCE
Tel:33 (0)1 69 15 49 88
Fax:33 (0)1 69 15 77 26
Email benoit.delatour@ibaic.u-psud.fr
Web http://www.namc.u-psud.fr

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