This video will illustrate the mouse eye and nucleation or removing the eyeball from the eye socket. The eyeball is bounded by the lower lid margin. Here is the medial or nasal canthus.
In the back is the temporal or lateral canthus. Above is the upper lid margin. Here's a surgical specimen.
The needle is pointing to the clear cornea. We can see a little bit of the lens poking past the iris. The iris is here in brown, covering the lens.
The most important surgical landmark is this white line, or the limbus will be making a puncture wound just behind it. The back half of the globe is behind the limbus and we're pointing to a muscle insertion. You can see the muscle lying against the globe here, and then the connective tissue that fills the orbit and includes the Arian gland.
And finally, this is the white long strip of optic nerve from a blunt dissection. Using a curved dressing forceps, customized with a sharp tip, begin by pulling apart the eyelids. This tends to prop toast the eye out and away.
Carefully use your forceps to go into the orbit and behind the eyeball, compress them gently behind the eyeball without squeezing it and pull forward. You should have been able to grasp the connective tissue and not the eyeball. If you turn the eyeball over to the side, you can see the orbital tissue along with the white optic nerve.
This is a second dissection just to show that again, you go in at the canthus, push the forceps behind the eyeball, grab the connective tissue, and pull it forward without depressing on the eyeball itself. Again, you can see the connective tissue in the long strip of white optic nerve. To make a puncture wound, grasp the eyeball by a muscle insertion and insert a needle behind the limbus right next to where you're holding the eye to stabilize it.
This will allow fixative to enter the eye. If you make the puncture far away from where you're holding the eye, the eyeball will be very unstable, and it'll be difficult to make that wound as long as you don't compress the eye or apply undue pressure. You can also grasp all the connective tissue just behind the eyeball itself.
Some fixatives don't require a puncture wound. In general, we put the eye in a five cc scintillation vial with at least three ccs of fixative to make sure the eye is submerged. For profusion fixed animals, a sharp dissection is necessary.
We use a curved calibri forceps and curved wescott scissors. Grasp the lower lid with the calibri forceps. Insert your scissors against the orbital bone, and begin cutting the connective tissue.
We use the curve away from the eyeball and avoid in sizing. Any of the eye will work all the way along the lower lid margin from one canthus to the other. At a certain point, most of the connective tissue will be removed, and you can slide your scissors behind the globe and pull it away.
Here's another example again, inserting the scissors into the orbit, but against the orbital bone rather than the eyeball and snipping away at connective tissue. This will begin to free the eyeball, which is like a basket ball in a basket. Use the forceps to retract the eyelids away and make gentle short cuts around the eyeball.
Here we grasp the connective tissue or a muscle insertion with the Collibra and finish cutting away the eyeball from the orbital connective tissue. After the eyeballs are suitably fixed, and if they need to be transported, we place some para film. We around the sation vial.
The packaging has to be in accordance with your university and government regulations. We typically place these into a container with some cotton To catch any fluid, this container has to be sealed very tight. This shipping container is then placed into a bubble wrap bag.
This is placed into a liquid laboratory shipping pack, along with any documentation.
