Scanning electron microscopy (SEM) was performed on 1 month old control (Pax6GFPCre:Caprin2+/cKO) and Caprin2 cKO/cKO mutant lenses as previously described (Scheiblin et al., 2014 (link)). Briefly, whole eyes were dissected without delay from control and mutant mice after euthanization and treated with a fixative containing 0.08M sodium cacodylate pH 7.4, 1.25% glutaraldehyde, 1% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA) for 3 hours. The lens were dissected from the eye and transferred to fresh cold fixative for 48 hours. The lenses were washed and the lens capsule along with a few layers of fiber cells were removed from one hemisphere of the lens in order to view the fiber cell ultrastructure. The peeled lenses were then dehydrated through an alcohol dilution series and hexamethyldisilazane (HMDS, Sigma, St. Louis, MO) dilution series (diluted in ethanol). Lenses were then subjected to sputter coating with gold/palladium for 2.5 min before imaging with Hitachi S-4700 Field Emission Scanning Electron Microscope (Tokyo, Japan). The analysis was performed on eight biological replicates for both control and mutants.