1. Trypsinize cells, count and plate cells in appropriate media at a density of 1 x 104 cells/well in a plastic 8 well chamber slide. Wells are pre-coated with ECM of choice (See slide coating protocol).

2. Incubate 3 hours to overnight at 37*.

3. Gently suction off media and wash 3x in 1x PBS.

4. Fix cells in 4% paraformaldehyde in PBS for 5-10 minutes at room temperature.

5. Wash 3x in PBS [-and then incubate in 0.1M glycine (in PBS) for 30 minutes at room temp-optional].

6. Wash 3x in PBS and then add primary antibody (diluted to appropriate concentration in 1% BSA in 1x PBS). Incubate in humid chamber at room temp. for 1.5 hours.

7. Wash 3x in PBS, and then add conjugated secondary antibody (Diluted 1:30 in 1% BSA in PBS). Incubate in humid chamber at room temp for 1.5 hours or overnight at 4*.

8. Wash 3x in PBS, add "Fade Slow" in glycerol and cover slip

9. Observe the cells under the fluorescent microscope.