Fixing tissues

Richard J. Shaw shawr at agresearch.cri.nz
Tue Nov 19 21:56:17 EST 1996

In article <Christine.Pechoux-1311961441310001 at mac-lr5-ottmann.ens-lyon.fr>,
   Christine.Pechoux at cri.ens-lyon.fr (Christine Pechoux) wrote:
>In article (Dans l'article) <32813601.5579 at forestry.ca>, Anthony, Pang:,
>apang at forestry.ca wrote (écrivait) :
>> I read somewhere that using formaldhyde in fixing tissues will
>> cross-link tissues in the sections. This may cause a problem in receptor
>> detection. Is this true? I am looking for the formula for Bouin-Hollande
>> 10% sublimate ( P. Brandtzaeg: Tissue preparation methods for
>> immunocytochemistry IN "Techniques in Immunocytochemistry Vol 1, edited
>> by G.R. Bullock and P. Petrusz, 1982"- I don't have this volume)
>> Please send formula to : apang at forestry.ca
>I some experiences in immunochemistry, and tissues fixation, so
>commercialy formaldhyde is not the best fixative existed. And
>Bouin-Hollande is really bad, and if you can avoid to use it !
>So paraformaldehyde fixative is is the most widely used fixative, either
>with or without the addition of glutaraldehyde.
>The paraformaldehyde concentration is between 1 to 4%, and high
>glutaraldahyde concentration (>0,5%) increased structural preservation,
>but also decreased the signal.
>So try both fixatives alone or mixed and goog luck

I have found that it is best to test as wide a range of fixative 
types/combinations/times to find the "ideal" combination for optimal 
structural and antigen preservation.  "Generally" of the aldehydes, fresh 
paraformaldehyde is best for optimal structural and antigen preservation.
I have found that usually for small antigens  eg hormones, glutaraldehyde 
hepls in the fixative.  For many cell surface antigens methanal/ethanol 
containing fixatives are all you can use.  Same applies to some monoclonals. 
In the end test test and test to find whats best for you purposes.  IMHO only

Richard Shaw                          Internet: ShawR at agresearch.cri.nz
Researcher (Science technician)
P.O.Box 40063, Upper Hutt

Technicians Do Most Things Better!!
	Always be Happy and Just Do It!!----------------------------------------------------------------------
Disclaimer: The above is a personal opinion and does not reflect the
            official view of AgResearch Ltd.

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