Cy3 labelled goat anti mouse igg

After the adult zebrafish had been exposed to the alcohol, their eyes and brain tissues were harvested and immediately fixed in 4% paraformaldehyde, equilibrated in 30% sucrose/PBS overnight and embedded in OCT. Twelve-mm-thick sections were mounted on gelatin-coated slides and air dried at 37°C for at least 2 h. The tissue sections were rehydrated with PBS, blocked with 20% NGS and 2% BSA in 0.3% PBS/Triton X-1 00 (PBST) for 1 hr, and incubated with primary antibodies overnight at 4°C. The following primary antibodies and concentrations were used: mouse monoclonal antibody Zpr-1 (1:200, University of Oregon) for labelling cones and mouse monoclonal anti-tyrosine hydroxylase (1:400, Millipore, Billerica, MA) for labelling DA cells. The interpretation of the neuroanatomy follows the adult zebrafish brain atlas [49 ] Immunoreactions were detected using Cy3-labelled goat anti-mouse IgG diluted to 1:400 (Millipore), and the sections were counterstained with a 1:1000 dilution of 4’, 6-diamidino-2-phenylindole (DAPI) (Sigma) to label the nuclei. The slides were examined with an Olympus BX51 light microscope (Olympus, Tokyo, Japan). The images were obtained by an Olympus CCD DP71 (Olympus) and processed using Adobe Photoshop CS (Adobe Systems, San Jose, CA).