Alexa fluor 488 conjugated goat anti rabbit

For whole mount lens anterior capsules, the anterior capsules were blocked and permeated with incubation of 1% Triton X-100 in 1% BSA for 1 h at RT. Primary antibodies diluted in PBS were added to immerse the tissues entirely and incubated for 12–24 h at 4°C. On the following day, capsules were washed with PBS containing 0.1% Tween20 (PBST) for 30 min on a rocking board, thereafter, incubated with appropriate secondary antibodies (diluted 1:200 with PBS) for 1 h at RT. After washing with PBST for 30 min, capsules were incubated with sufficient volume of DAPI for 10 min to stain the nuclei. Finally, the whole anterior capsule was placed flat on a microscope slide using forceps under a dissecting microscope. The residual PBS on the slide was removed, and the capsule was covered with a coverslip after adding a drop of anti-fade mounting medium. The slides were stored at −20°C in the dark until examination using a fluorescence microscope and a LSCM within a few days.
Primary antibodies against α-SMA, collagen type I (Col I), and vimentin were purchased from Abcam (Cambridge, UK). Alexa Fluor 488-conjugated goat anti-rabbit and Alexa Fluor 555 conjugated donkey anti-mouse secondary antibodies were purchased from Cell Signaling (Danvers, MA).