Materials

• Fibronection from Corning

15 ug in 300 uL PBS per slide (surface area is 1.7×1.7 cm = 2.89cm2, corning protocol show 1～5 µg/ cm2, 2.89 cm2 x 5 ug = 14.45 ug)

• 5% BSA/PBS

1. Priming with ethanol

Please check “1. Priming with ethanol” in printed user guide

2. Wash with PBS

Please check “2. Wash with DPBS” in printed user guide

3. Add Fibronectin

1. Add 300 uL FBN solution. Surface is hydrophobic. Rock gently to ensure even coating of the surface.
2. Aspirate 50 uL(25 uL from both ports)
3. Incubate >1 hr at room temperature with lid (ensure solution is coverying surface)
4. Aspirate 250 uL of solution from side ports
5. Add 2 mL of DPBS
6. Aspirate 2 mL of DPBS
7. Repeat 5-6 You will find that surface is hydrophilic.

4. Blocking

1. Add 300 uL 5% BSA/PBS solution.
2. Aspirate 50 uL(25 uL from both ports)
3. Incubate >1 hr at room temperature with lid (ensure solution is coverying surface)
4. Aspirate 250 uL of solution from side ports
5. Add 2 mL of DPBS
6. Aspirate 2 mL of DPBS
7. Repeat 5-6

5. Cell Loading

Please check “4. Cell loading” in printed user guide

This shows PC3 cells, 30 min after cell loading. Cell shape is typical flattened morphology.

This show DAPI and phalloidin ifluor 488 staining after 1 day culture.

This shows A549 cells in FBN coated Hex20 3 days after loading, staining with DAPI, CellBrite(Green) and phalloidin(Red).