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Is it necessary to coat VisiSens sensor foils with cell adhesion promoting proteins, and how do I coat them?
When using the O2 Sensor Foils SF-RPSu4 with the optical isolation white (OIW) layer as cell substrate, we recommend coating (according to the instructions below) to promote cell adhesion. For SF-RPSu4 without OIW and SF-HP5R coating is not mandatory.
The table below lists all cell lines that have been tested for their adhesion to SF-RPSu4 and SF-HP5R:
|Sensor Material||Cell Type||Confluent Cell Layer (after 24 hours)||Coating|
|O2: SF-RPSu4-OIW||MDCK II||No||Recommended|
|O2: SF-RPSu4 without OIW||MDCK II||Yes||Not mandatory|
|O2: SF-RPSu4 without OIW||NRK||Yes||Not mandatory|
|O2: SF-RPSu4 without OIW||BAEC||Yes||Not mandatory|
|O2: SF-RPSu4 without OIW||U-373||Yes||Not mandatory|
|pH: SF-HP5R||MDCK II||Yes||Not mandatory|
- Attack the sensor foils to your cell culture vessels (e.g. Petri dish, well plate or cover slip) with the provided silicone adhesive, high vacuum grease, or use self-adhesive sensor foils. When using a silicone adhesive, allow for 24 h of curing. Then rinse the surface with water or buffer to remove acetic acid liberated during curing.
- Apply coating solution for 1 h at the temperature indicated in the table below. Make sure the coating solution covers the sensor foils completely. Please note that hydration of the pH sensor foil (hydrogel) takes extra time that should be added to the total time for coating.
- Aspirate the coating solution after 1 h and seed the cells.
|Coating Material||Concentration||Incubation Parameters|
|Culture medium||cont. 5 % (v/v) serum||1 h, 37 °C, 5 % CO2|
|Fetal Calf Serum (FCS)||pure||1 h, 37 °C, 5 % CO2|
|Fibronectin (from human plasma)||25 µg/mL||1 h, room temperature|
|Gelatin||0.5 % (w/v)||1 h, 37 °C, 5 % CO2|
|Poly-L-lysine||100 µg/mL||1 h, room temperature|