9
►
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.
The PDMS (silicone) chambers
are very hydrophobic with two
methyl-bases on the surface.
Cells adhere to the fibronectin
or collagen coated chambers
via integrins. This form of cell
adhesion is very different
compared to attachment of
cells to plastic or glass dishes
where the surface of plastic or
glass is charged, resulting in
non-specific binding.
If the cells are having difficulty
attaching to the coated
chamber or easily detached
upon stretching, treat the
stretch chamber with a higher
concentration of fibronectin or
collagen, for overnight.
