To begin, obtain anesthetizing and analgesic agents, dilation drops, sterile disposable scalpel, desired types of forceps, needles, syringes, and balanced salt solution or BSS. Sterilize the surgical instruments. Prior to anesthesia, administer one drop each of 1%atropine and 2.5%phenylephrine to dilate the pupil of the eye undergoing surgery.
Using a sterile 26 gauge half inch needle, prepare a syringe with sterile BSS. Then attach a sterile one inch, 27 gauge blunt dispensing tip bent at 45 degrees to the syringe. Following the mouse's anesthesia, perform a toe pinch to confirm anesthesia depth ensuring pupil dilation reaches two millimeters.
Move the anesthetized mouse to the stage of a compound dissecting light microscope with a heating pad. Using a sterile surgical scalpel, make a one to 1.5 millimeter incision in the center of the cornea, ensuring that the anterior lens capsule is also incised. Gently expel one drop of BSS above the cornea to wet the surface for further manipulation.
Now, insert the dispensing tip through the central corneal incision into the lens capsule incision, and gently move the needle in a circular motion while slowly releasing BSS to expel the lens fiber mass. Once the fiber mass adheres to the dispensing tip, gently remove the dispensing tip from the inside eye. Clean it with a sterile towel to remove fiber cells and repeat until all lens fiber cells and lens nucleus are removed.
Rinse the lens capsule with sterile BSS to remove any remaining fibers. Observe the translucent or glassy appearance to confirm that the capsule is inside the eye. To close the corneal incision, first, hold one corneal flap using forceps in the non-dominant hand, and push the needle with nylon thread through it.
Then push it through the next corneal flap. Place one square knot stitch in the central part of the incision and trim the excess thread. Place the bent needle through the corneal gap either above or below the recently placed suture and gently expel BSS to reinflate the eye.
For postoperative care, immediately apply one drop of 0.5%erythromycin ophthalmic ointment directly above the suture covering the entire incision. Administer buprenorphine intraperitoneally. Allow the mouse to recover from anesthesia in a cage placed on a heating pad to optimize thermal support and monitor until it recovers.
