Figure 1.

Retinal dissection from an enucleated mouse eye

(A) An enucleated mouse eye submerged in PBS.

(B) Grasp the sclera or a stump of residual extraocular muscle with Dumont Forceps #5 and secure the eye in position. Create a puncture in the sclera across the globe using a 30-gauge needle.

(C) The dotted circle indicates the scleral puncture.

(D) Grasp the anterior sclera at the edge of the puncture with the Dumont Forceps #5 and create a circumferential incision parallel to and just posterior to the limbus with Vannas scissors.

(E) Remove the cornea from the posterior eye cup by holding the eye at the limbus with forceps.

(F) Use a second pair of forceps to hold the posterior cup at the optic nerve to create traction.

(G) While securing the optic nerve with forceps, pull away the crystalline lens with a second pair of forceps.

(H) An inside view of an isolated posterior cup shows the neuroretina surrounded by retinal pigmented epithelium and sclera.

(I) Use a pair of forceps to grab the scleral edge and begin folding back the sclera from the retina.

(J) Continue to use both pairs of forceps to peel back the sclera from the retina and work circumferentially around the limbal edge.

(K) Evert the sclera to prolapse the retina.

(L) Continue to gently separate the retina from the sclera.

(M) The scleral cup is now completely everted and only attached to the retina at the optic nerve head.

(N) Secure the sclera with forceps and create four radial incisions within the retina.

(O) Even with relaxing cuts, the retina still retains a folded shape at this stage.

(P) Make the final radial incision in the retina. Relaxing incisions are 90 degrees apart.

(Q) Detach the retina from the sclera by axotomizing RGCs at the optic nerve head with the Vannas scissors.

(R) Aspirate the free-floating retina using a P1000 pipette with a widened tip.

(S) A view showing the retina inside the P1000 pipette tip.

(T) Transfer the retina onto the center of a culture insert and position the retina with the inner retina facing up.

(U) Aspirate the excess PBS around the retina. The retina edges will start unfolding and flattening.

(V) An alternative method of unfolding the edge is by using a bent vascular probe to engage the overlying fluid and use surface tension to draw the folded retina peripherally.

(W) Use the surface tension between the vascular probe and the folded surface to flatten out the retina.

(X) The flatmounted retina on the culture membrane. Scale bar, 2 mm.