mouse retinal dissection - removing the retina (Dec/31/2009 )

Hello all,
I recently began angiogenesis studies in the mouse retina and am having a fair amount of difficulty dissecting out the retina from the rest of the eye. I have looked exhaustively throughout the internet and through the literature for some guidance but have found relatively nothing. I am well beyond the point of frustration. Was hoping for any advice, tips, direction, etc.... Thanks in advance

gd20 on Dec 30 2009, 11:21 PM said:

I do not envy you. I used to have to do that with zebrafish.

I can't really explain the technique in detail (I'd have to show you which I obviously can't do), but I can tell you this much:

1) Be sure to cut the optic nerve prior to enucleation or you risk severely tearing the retina
2) Remove the lens using two sets of forceps - one to hold down the eye, the other to pull the lens off. Grab the lens from the front and the back with the set of forceps and pull up and to the side
3) Using either two sets of foreceps or one set and your finger, squeeze the vitreous out of the eye through the space left by the lens.

At this point, if you're not worried about getting the whole retina you should be able to just cut off the front of the eye to remove the cornea and you'll be golden.

Hope that helps!

-Carlton

This is my protocol. Its better to understand if someone could show it to you atleast once.


''Puncture the eye. Cut out the cornea. cut around the eye at the limbus, very close to the limbus (retina is attached to the eye cup into 2 places: at the limbus and the optic disk), to release the retina. After the cut around the limbus, place a forceps in the optic disk to fix the eyecup, now gently with the other forceps (preferabbly blunt tip), go around the edges to free the retina from the eyecup. Now the retina is fixed only at the optic disk, with the forceps used to fix the optic disk, pressurise the area so that the optic nerve is damaged and frees the retina or go from beneath the retina and hold the optic nerve and try to pluck the retina gently.''

good luck