Using Coated Glass Coverslips For Cell Culture

(Source: D. Frost and D. Sepich from Zebrafish Book 5th Edition)

Zebrafish embryonic cells attach well to glass coated with laminin or to plastic.  They do not attach well to bare glass or glass coated with fibronectin. They attach somewhat to 0.2 mg/ml gelatin coated glass.

1. Sterilize glass coverslips (22 mm x 22 mm) by dipping in ethanol and flaming.
2. Place a drop of 10 μg/ml laminin (Boehringer Mannheim) in sterile PBS and leave for 45 minutes.
3. Rinse thoroughly with sterile PBS or dH₂O.
4. Place a drop of cell suspension (cells from 1-4 embryos) onto a 0.5-1.0 cm spot in the center of the coated glass coverslip in a Petri dish and allow the cells to attach for several hours or overnight. 
5. Flood the dishes with growth medium.