Histology Fixative

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msvnathan
msvnathan's picture
Histology Fixative

Bouin’s Fluid

Picric Acid, saturated aqueous - 75ml
Formalin (37-40% formaldehyde) - 25ml
Glacial Acetic Acid - 5ml
Combine all three to make up fixative.

Hopwood, D. (1990). Fixation and fixatives. In Bancroft, JD. et al. (eds). Theory and practice of histological techniques. Churchill Livingstone, New York, p21-42

Note:
After fixing the tissue if the tissue is big in size it can be fixed 24hrs first and then taken out cut into pieces and again fix in the same solution for another 24hrs. After fixation the sample should be washed and can be kept in 70% alcohol (30% then 50% and finaly 70%) till it used for further work to avoid hardening of tissue (if kept in fixative itself tissue will become hard)

Guy Sovak
Guy Sovak's picture
Hi ,

Hi ,
Thanks for the protocol,
For your knowledge, for the couple of years Bouins Fixative is rarely used as it is highly toxic and carcinogenic.
Better to use the regular Natural buffered Formalin (10% NBF).
Guy

msvnathan
msvnathan's picture
Hi, Thanks for the

Hi, Thanks for the information. For Immunohistochemistry work mostly we are using Buffered Formalin, for H&E staing we used Bouins Fixative.

Vaithinathan.

peter36
peter36's picture
Bouins fixative is great for

Bouins fixative is great for nuclear detail for say testicular bx or bone marrows. The problem being as you say harding of tissues, the need to wash the picric acid completely out of the tissue in say 70% alc or you will staining properties wwith haematoxylin plus you will have a problem with your processing schedule which usually starts with formalin(water based). you can use 10% formalin for most of your work with good results and keep the bouins for above tissue.
pete

peter36
peter36's picture
<p>Bouins fixative is great

<p>Bouins fixative is great for nuclear detail for say testicular bx or bone marrows. The problem being as you say harding of tissues, the need to wash the picric acid completely out of the tissue in say 70% alc or you will lose nuclear staining properties with haematoxylin plus you will have a problem with your processing schedule which usually starts with formalin(water based). you can use 10% formalin for most of your work with good results and keep the bouins for above tissue.</p> <p>pete</p>