►
Fibronectin Coating
Preparation of fibronectin solution:
1. Dilute human or bovine fibronectin to a final concentration of 50 to 100 µg/ml in
Phosphate Buffered Saline (PBS)
Coating with fibronectin solution:
1. Pour 3 ml of the fibronectin solution into each strain chamber
2.
Incubate at 37
°C for more than 30 minutes
3.
Aspirate the fibronectin solution. If coating is successful, water will not be repelled after
removing the
fibronectin solution.
4.
The liquid solution can be used to coat 3 or 4 chambers before discarding.
►
Gelatin Coating
Preparation of gelatin solution:
1. Add gelatin powder to PBS at a concentration of 2%
2. Autoclave the mixture to dissolve and sterilize
Coating with gelatin solution:
1. Pour 3 ml of the gelatin solution into each strain chamber
2.
Incubate at 37
o
C for more than 30 minutes
3. Aspirate the gelatin solution. If coating is successful, water will not be repelled after
removing the
gelatin solution.
4. The liquid solution can be used to coat 3 or 4 chambers before discarding.
►
Collagen Coating (Cellmatrix 1-C, P, Type 3 or 4)
Preparation of collagen solution:
1. Combine 1 part collagen to 10 parts HCL, pH 3, in a sterile tube
Coating with collagen solution:
1. Coat chamber with a thin layer
2. Aspirate excess
3. Dry in biological safety cabinet at 25°C or below. The chamber can be stored at the
same
temperature.
4. Wash the chamber twice with culture medium.
5. If coating is successful, water will not be repelled.
Important:
If cells are having difficulty attaching to the freshly coated chambers or are
easily detaching upon stretching, treat the chamber with a higher concentration of the
