Hi all,
I have perfused mice with 1% PFA and post fixed eyes in 1% PFA then removed cornea and lens and put the eye cup (containing retina) in OCT for cryosectioning. Th problem is that the protocol requires the retina to be fixed for short time and that makes it challenging to embed it in OCT as the retina is still soft. I am thinking to sections the whole eye without removing cornea and lens. Have you tried sectioning mouse eyes with lens on? Do you have any recommendation to avoid retinal breakage whole I remove the lens for OCT sectioning?
Thank you
Assraa
Another option is to inject fixative into the vitreous through a pars plana approach: this works well when I am fixing eyes in gluteraldehyde for electron microscopy of the retina.
OCTs machines are constructed for normal anterior segements with a cornea, lens, and pupillary system. I suggest either to take images in narcotized animals or eyes before resection of the anterior segment, as discussed by Dr. Williams.
You may try
i- 4% bufferized paraformaldehyde for perfusion.
ii- Increase the perfusion time.
iii- Post fix in 10-40% sucrose gradient solutions and finally freeze it in oct compound.
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