Dear readers,
Please share your experiences or advise me on the topic above.
The Bouin's fluid is often preferred over formalin for fixation of the testis. Based on your experience or knowledge, what are the best practices to achieve the best quality of the H&E stained tissue sections for histomorphological studies?
Also, how to preserve the tissue fixed with Bouin's solution for an extended period with the least effect on the quality of the preserved tissue sample?
What is the accepted tissue processing protocol for the testis tissue fixed with Boiun's solution.?
Thanks and Regards,
Hello Mahesh Shriram Deokar ,
Fixation with Bouin's Solution (often preferred for HES or H&E and IHC analyses) followed by microtome sections can be difficult to perform depending on different parameters:
• The fixation time of the tissue (if it is overfixed, the tissue will be fragile and "breakable"), considering the size of the tissue.
• The Bouin's Solution-fixed tissues have to be store at 4°C in 70° alcohol O/N and then embedded in paraffin.
• The age of the blade which cuts the block (it is sometimes necessary to have a recent blade but also of a well defined nature for certain tissues)
• The nature of the paraffin (some tissues require a special high quality paraffin)
• The ambient & block temperature (the tissue sample and the paraffin are cut much better when they are cooled on a plate provided for this purpose or beforehand with ice cubes). In this month of January, it may beok in your lab...
Hoping that will be useful for your testis sample analyses.
For more informations, please see the "Histological analysis" of our lab's article : Article Assessment of the optimal vitrification protocol for pre-pub...
In addition, please see this topic : https://www.researchgate.net/post/How-to-remove-the-yellow-colour-in-tissue-processing-if-tissue-is-fixed-in-Bouins-solution
Ludovic Dumont, Ph.D.
Hi Dr. Dumaont,
Thank you very much for your valuable advice. I will update you as I will be processing some tissue samples soon.
Thanks again and Regards,
Fixation time would depend on the size of the tissue. Generally 24-72 hrs would be acceptable. With Bouin's it's best not to keep the tissue in fixative for much longer than this; generally the tissue will be transferred to 70% ethanol until ready for processing (which should be done promptly, as ethanol will harden the tissue and make it brittle for sectioning). Processing is done in the same manner as you would use for formalin-fixed specimen, except that you would probably start the cycle in ethanol rather formalin.
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