Does anyone know how to soften the eyeball for histology in paraffin technique? Currently, we use a solution of 4% phenol in 70% ethanol before we put slices to the tissue processor, but they are still hard to cut.
I'll be grateful for the help.
The following article could help you. The authors use concentrated liquid phenol to soften the lens.
Arko-Boham B, Ahenkorah J, Hottor BA, Dennis E, Addai FK. Improved method of producing satisfactory sections of whole eyeball by routine histology. Microsc Res Tech. 2014 Feb;77(2):138-42. doi: 10.1002/jemt.22320. Epub 2013 Nov 18.
I work with ocular pathologies in MA and have obtained good results in fixation with Davidson solution.
Place the intact globe in Davidson for 24 hours, after making the cut, and maintained for 12 hours. Transferring to 70% alcohol.
Davidson solution:
Glacial acetic acid: 100 ml
Ethyl Alcohol 95% 300mL
Neutral buffered formalin 10%: 200 ml
Distilled water: 300 ml
Good luck!
Hi Agnieszka, I have not done this kind of processing for a long time and then it was for ocular melanoma, but my interest was in best preservation of the tumour, less in structures of the eye. I checked the literature - Frederic gave you link to the latest paper. It pays to do a Google search - I have found that a similar question was asked by another ResearchGate member over a year ago and received quite a few answers. https://www.researchgate.net/post/Could_someone_share_a_protocol_for_processing_the_eyeball_for_histological_studies.
There is a lot of interesting facts there about fixation and processing (esp. look at the PPoint handout). It all depends what you want to achieve - if it is histology only and you need to have lens intact - then phenol is needed, but if you plan immunohistochemistry - then contact me directly, if you wish. Good luck with your work.
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