Materials and Reagents

Note: All reagents may be maintained at room temperature.

1. FVB mouse (5-10 weeks old)
   
2. Isoflurane (Abbott Laboratories, catalog number: 5260-04-05 )
   
3. Proparacaine hydrochloride ophthalmic solution (0.5%) (Bausch & Lomb, NDC: 24208-730- 06 )
   
4. Fluorescein sodium and benoxinate hydrochloride ophthalmic solution (0.25%/0.4%)  (Akorn, NDC: 17478-640-10 )
   
5. Weck-cel cellulose eye spears (Medtronic, catalog number: 0008680 )
   
6. NaOH solution (see Recipes)
   
   

Chemical injury

7. Sterile water
   
8. 1x sterile phosphate buffered saline buffer (1x PBS)
   
9. Sodium hydroxide (NaOH) 1.0 N (normal) solution (Sigma-Aldrich, catalog number: S2770 )
   
10. Fluorescein solution (see Recipes)
    

Equipment

1. Heating pad for mouse
   
2. Algerbrush II with 0.5 mm Burr (Katena, catalog number: K 2-4900 )
   
3. Trephine with handle (1.5 mm) (Beaver-visitec, catalog number: 9748 )
   
4. Alcohol swabs - isopropyl alcohol 70% (BD, catalog number: 326895 )
   
5. Filter paper (Thermo Fisher Scientific, catalog number: 09-795AA )
   
6. Ear punch (Roboz, catalog number: 65-9902 )
   
7. Forceps (Dumont #5)
   
8. Pipette with tips (1 ml)
   
9. Timer
   
10. Stereo microscope (for scratch/epithelial injury, need filter set to visualize GFP; for both injuries, camera attachment is optional) (Leica, catalog number: MZ16F )
    
11. Anesthesia machine with nose cone attachment appropriate for mice (Summit Anesthesia Solutions, catalog number: AS-01-0007 )
    

Procedure