Re: Golgi methods for fixed brain

From:"J. A. Kiernan" <jkiernan@uwo.ca>


On 24 Apr 2001 danand24@123india.com wrote:

> I am Anandh,Doing research in neurophysiology.My work is on Aluminium 
> toxicity and morphological studies.I have to use golgi staing for my
> work and i like to know that is there any golgi method for formalin
> perfused brain.I have gone through your letter to histonet which is
> avilable in web.I am very much interested in knowing that techneque.
> So please give me some refreances so that i can go further forward 
> in my work.I will be grateful to you .Thanking you.

Here are some references, with brief notes. They are not in any 
particular order. My experience is limited to methods that use
the Golgi-Cox mixture as the primary fixative. Kolb & McLimans
(1986) is a good modern method of this type. If you have a vibrating
microtome (vibratome) you should get nice 100um sections for
looking at dendrites.
 
 BERTRAM, E. G. & IHRIG, H. K. 1959
 Staining formalin-fixed nerve tissue with mercuric nitrate stain.
 Stain Technology 34, 99-108.

 Staining formalin-fixed nerve tissue with zinc chromate followed by sat.
 mercuric nitrate (pH 5.5-6; critical). A Golgi-Cox type of
 method. (Tissue fixed by pulsating perfusion with formal-acacia, 
 pH 7.0)


 GONZALEZ-BURGOS, I., TAPIA-ARIZMENDI, G. & FERIA-VELASCO, A. 1992
 Golgi method without osmium tetroxide for the study of the central
 nervous system.
 Biotechnic & Histochemistry 5, 288-296.

 A 3-day Golgi technique. Formalin perfusion fixn of brain. Dichromate
 with formalin, acetic acid, wash, AgNO3. No need to protect surface.


 GRANDIN, T., DEMOTTE, O. D., GREENOUGH, W. T. & CURTIS, S. E. 1988
 Perfusion method for preparing pig brain cortex for Golgi-Cox
 impregnation.
 Stain Technology 63, 177-181.

 A dissecting fixative (phenol, formald etc), prior to a Golgi-Cox
 procedure.

 
 GREER, E. R. & JEN, C. K. 1990
 Modified Golgi method for whole rat brain.
 Stain Technology 65, 155-157.

 Buffered formaldehyde fixation before Golgi-Cox procedure. 150 Ám
 cryostat sections; Li(OH)2 & KNO3.


 KOLB, B. & McLIMANS, J. 1986
 Cryostat sectioning of Golgi-Cox tissue.
 Stain Technology 61, 379-380.

 Primary fixation (after saline perfusion) in dichromate-mercuric
 chloride, 14- 20 days, dark. Cryostat sections treated with ammonia, 
 Kodak photo fixer (1/7).


 FISH, P. A. 1895
 The use of formalin in neurology.
 Transactions of the American Microscopical Society 17, 319-330.

 Review (31 refs) followed by some original observations. Recommends Water
 2000, Formalin 50, NaCl 100, ZnCl2 15. Fix brain 7-10 days. Perfuse 
 vessels & inject cavities if possible. Post fixation in 2.5% formalin, 
 or various mercury, picric, dichromate solutions. Also various 
 Golgi-type methods; formalin- dichromate mixtures must be kept in 
 darkness.

I hope these will help you to choose a suitable technique. 

----------------------------------------
John A. Kiernan
Department of Anatomy & Cell Biology
The University of Western Ontario
London,  Canada   N6A 5C1
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan




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