Cryoimmuno electron microscopic analysis of wild type and R49C
neo knock-in lenses. Lenses expressing either wild type or heterozygous R49Cneo/R49Cneo αA-crystallin were labeled with an antibody to αA-crystallin and analyzed by cryoimmuno electron microscopy. (A) Wild type lenses showed linear fiber cell membranes. Immunogold labeling (black dots) showed that αA-crystallin was restricted to the cytoplasm. (B) Heterozygous WT/R49Cneo αA-crystallin lenses had several gold particles on the fiber cell membranes. Furthermore, the linearity of lens fiber cell membranes was lost, and significant gaps between cells were apparent.