Figure 2. Whole mount electro-immunofluorescent staining with anti-keratocan. Wild type mouse corneal stroma was labeled by goat anti-keratocan Alexa555 conjugate (red) and nuclei were stained with DAPI (blue) as described in Methods. The red fluorescence is only detected in corneal stroma (A). No red fluorescent signals are found in the keratocan knockout mouse cornea by goat anti-keratocan Alexa555 conjugate (B). The wild type mouse corneal stroma is not labeled by non-immune goat IgG Alexa555 conjugate (C). Conventional whole mount fluorescent immunostaining with anti-keratocan-Alexa 555 conjugate showed that the antibody couldn’t penetrated into the deep corneal stroma and exhibited non-specific binding on the surface of the corneal epithelium and endothelium (D). Scale bar: 10 μm.