I have a good experience of special fixative working very good in samples of a donkey testis(I attached three published article from my master thesis using Worbel Moustafa fixative) .
I want help for the best fixative for the eye and the best way to apply the fixative by injection between sclera and cornea by syringe or by making a small opening and injection the fixative slowly inside the eye. and left eyes in the fixative overnight at 4'C- Then wash in 0.1 M PBS 3 times to remove fixative.
I have also the attached file for eye fixation(two files).
I want help for the best fixative for the eye and the best way to apply the fixative by injection between sclera and cornea r by making a small opening and injection the fixative slowly inside the eye.
I wish to receive the answer. I want good fixation of the retina
I've worked a lot with fixation of rabbit eyes and have been using davidssons fixative for 48h before 4%PFA. That fixation has turned out well for histopathology.
Perhaps that might work?
I opened the eye via longitudinal section in cornea and i removed the lens and fluids inside the eye. Then I fixed it in 4% PFA overnight at 4 degree, it works well. I made that in eye of large animals but for fish fries I fixed the whole fish in 4% PFA overnight at 4 degree and the retina easily observed.
I perform the same as Mona N. Hussein . Just immersing the eyes in PFA 4 % and without injection of the fixative. You can also use Clarke´s solution if you want to section the eye with a paraffin microtome. However, you can fix them with PFA 4% and section in paraffin too. Tissue morphology is better after paraffin sectioning. Also a warning: Some antibodies don´t work in paraffin sections, and you have to remove autofluorescence of paraffin.
I know another one: you can fix in glutaraldehide, but I haven´t tried it before.
Hope this info is understandable and useful.
If you need the whole eye it is always better to make corneal cuts with a small surgery knife to allow PAF 4% penetration within the eyeball. A good fixative for paraffin sectioning is Bouin´s however it may reduce immunoreactivity to primary antibodies. To analyse the retina and RPE, choroid and sclera, I have always prepared eyecups (the eyeball after excising the cornea and lens and in rodents also the vitreous with small dissection forceps) and then immerse them (eyecups) in PAF 4% for at least one or two hours at RT.
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