Hippocampal slice culture protocol:

Materials and buffers to prepare:

1.Dissection solution: Gey's balanced sal solution supplemented with D-glucose to a concentration of 6.5mg/ml. (pH=7.1, 340m OSM)

2.Culture medium:

Basal Medium Eagle Medium                          50%

Earle's  Balanced Salt Solution                       25%

Horse serum                                                  15%

L-glutamine                                                    1mM

(pH=7.1, 320m OSM)

Add before each use:

Glucose, to a volume ratio of 45%

PS(0.5ml/50ml for PS of 5000U/ml)

Procedure:

1. Kill the post-natal rat following the experimental animal rules.

2. Dissect the brains under dissection microscope and take the hippocampus out.

3. Cut the hippocampus into 40μm slices using microtome.

Immunohistochemical staining:

BrdU, NeuN