Organotypic Culture of Developing Striatum - Drugs of Abuse: Neurological Reviews and Protocols - page 398

14. Methanol.
15. Triton X-100 (Sigma, St. Louis, MO).
16. Normal goat serum (GIBCO BRL, Grand Island, NY).
17. Normal rat serum (GIBCO BRL, Grand Island, NY).
18. Azide.
19. Biotinylated goat anti-rabbit IgG antibody (Vector Laboratories, Inc., Burlin-
game, CA).
20. ABC kit (Vector Laboratories, Inc., Burlingame, CA).
21. 3,3 -Diaminobenzidine tetrahydrochloride (DAB, Sigma, St. Louis, MO).

22. Nickel ammonium sulfate.
23. Scissor.
24. Forceps.
25. Scalpel.
26. Dumont no. 5 forceps.
27. Spatula.
28. Krazy Glue Pen (Borden Inc., Columbus, OH).
29. Razor blades.
30. 35-mm tissue culture dish (Falcon, Becton-Dickinson and Company, Franklin
Lakes, NJ).
31. Six-well culture plates (Falcon, Becton-Dickinson and Company, Franklin
Lakes, NJ).
32. 30-mm culture plate inserts (Millicell-CM, PICM 03050, Millipore, Bed-
ford, MA).
33. Water bath.
34. Vibratome (Ted Pella Inc., Redding, CA).
35. Dissecting microscope.
36. CO 2 incubator.

3. Methods
3.1. Preparation of Organotypic Culture of Striatum
The organotypic slice culture method of Stoppini et al. (1) was adapted for
use with modifi cations.

1. Sterilize all surgical instrument by placing the instrument in a beaker containing
70% alcohol.

2. Prepare 4% low melting temperature agarose in Gey's Balanced Salt Solution
(GBSS). Boil the solution to dissolve the agarose ( see Note 1 ). Once agarose is
dissolved, keep the bottle containing agarose in a 45°C water bath to maintain
the agarose in liquid form.

3. Place rat pups on ice for 5-10 min for hypothermia anesthesia.

4. Clean the pup's head with 70% alcohol.

5. Decapitate the head with a scissor.