Axiocam 506 monochrome

After the exposure, the same sagittal sections used for autoradiography were fixed in cold acetone (approx. -15 °C), quickly dried under a fan and blocked with 20% normal goat serum for 1 hour at RT. After disposing the goat serum, the tissues were incubated with a Goat anti-human IgG [H+L] Cross-Adsorbed Secondary HRP-Antibody (Invitrogen; 0.4 µg/mL, 1:2000) at RT for 1 hour under dark conditions. After disposing of the antibody solution, the tissue was washed three times with 0.05% Tween-20 in PBS for 5 minutes each, followed by a final wash step with deionized water (dH₂O) for 5 minutes. Subsequently, the tissue was incubated with a 0.125% freshly filtered Thioflavin S solution at RT for 8 minutes under dark conditions. After disposing the Thioflavin S solution, the tissue was washed for 3 minutes each in the following order; 2x80% EtOH, 1x90% EtOH, 3xdH₂O. The tissues were mounted with ProLong™ Gold Antifade Mountant (Invitrogen™, P36930). Images of the stained sections were taken with a fluorescence microscope (Zeiss Axio Observer with a Colibri 7 LED light source and an Axiocam 506 monochrome camera) and equally processed using the Zen blue software Version 3.4.