Day	Action
Before starting	Prepare astrocyte culture and freeze cells down. One batch of astrocyte culture can usually support several months of hippocampal cultures.
Week 1 Monday	Thaw astrocytes.
Week 1 Tuesday	Feed astrocytes.
Week 1 Friday	Feed astrocytes.
Week 2 Monday	Feed astrocytes. Rinse coverslips in H2O, then soak in nitric acid for 24 hr.
Week 2 Tuesday	Rinse coverslips 5x. Dry sterilize coverslips.
Week 2 Wednesday	Place paraffin dots on coverslips. Coat coverslips with poly-L-lysine.
Week 2 Thursday	Rinse coated coverslips in sterile H2O, then incubate coverslips in plating media overnight at 37 °C. Change media on astrocytes to NB27.
Week 2 Friday	Hippocampal dissection and plating.