Acetylcholinesterase Tissue Staining

The last staining step was targeting acetylcholinesterase, which is an enzyme that catalyzes the hydrolysis reaction of acetylcholine. We did this similarly to the parvalbumin staining, with a few tweaks to the protocol. First, the solution was prepared with a phosphate buffer and hydrogen peroxide. Then, a maleate buffer was used, which caused a slight peach color during incubation. TRIS Buffer rinses were followed by an incubation period , which was then followed by hydrogen peroxide and Tris Buffer rinses. The tissues from the three rat brains successfully reached a dark purple to black color, and were then mounted on gelatinous slides. These slides as well as the parvalbumin stained tissue slides will be subsequently analyzed by Dr. Burk to assess the effectiveness of  anti-ChAT saporin as a novel immunotoxin in the thalamic reticular nucleus.