Cataract surgery was modeled in mice using a modified extracapsular lens extraction technique (ECLE) based on previously published techniques.32 (link)–34 (link, link) Adult mice were anesthetized with 80 mg/kg ketamine and 5 mg/kg Xylazine. One eye of each mouse was dilated using several drops of topical phenylephrine and tropicamide. A 1- to 1.5-mm central corneal incision was made using a disposable ophthalmic knife. Following reinflation of the anterior chamber with an ophthalmic viscoelastic agent, a similarly sized incision was made in the anterior capsule. A viscoelastic cannula was used to instill saline into the capsular space to hydro-dissect the lens fiber mass away from the capsule. Angled jeweler forceps were used to remove the lens mass by applying gentle pressure near the equator of the eye. After the lens mass was expelled, careful irrigation of the capsule was performed to remove any residual lens material (in particular, the lens cortex). A viscoelastic agent was then injected into the capsule and anterior chamber to reinflate the eye and maintain its structural integrity postoperatively. The corneal incision was closed using 10-0 nylon sutures. Animals were euthanized 5 days postoperatively and lenses removed from both the surgical eye as well as the contralateral eye, which served as an experimental control. For analysis, whole eyes from ECLE experiments were preserved for immunofluorescence. In some cases, capsular bags were microdissected from the eye and used for isolation of RNA for PCR measurements. Surgery, euthanasia, and dissection were performed in parallel for each experiment to ensure consistency and comparability. Furthermore, the surgeon was blinded to the genetic strain of each mouse during operations. For experiments involving treatment with Sorbinil ([4S]-6-Fluoro-2,3-dihydro-spiro[4H-1-benzopyran-4,4′-imidazolidine]-2′,5′-dione), mice were injected intraperitoneally with 10 mg/kg at the time of surgery followed by twice daily injections on postoperative days 1 through 5. Sorbinil was provided by Pfizer Central Research (Groton, CT, USA). Vehicle controls contained 1X phosphate buffered saline.