8
Preparation of Silicone Chambers
Before using the chambers, they should be sterilized then coated with a cell adhesion
matrix. The coating procedures below can be adapted for use with other matrices,
such as Elastin, Pronectin, and Laminin.
Sterilize chambers in an autoclave for 20 minutes at 120°C.The silicone chambers can
withstand temperatures up to 180°C. Use of an autoclave is preferable. However, if
an autoclave is not available, the chambers may be sterilized by submerging in 70%
ethanol, rinsing with water, then drying in a sterile environment.
Place the sterile silicone chambers in a Petri dish in preparation for coating.
►
Fibronectin Coating
Preparation of Fibronectin solution:
1. Dilute human or bovine Fibronectin to a final
concentration of 50 to 100 ug/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
o
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.
PBS(per liter):
NaCl
8.00 g
KCl
0.20 g
Na
2
HP
4
(anhyd.)
1.15 g
KH
2
PO
4
(anhyd.)
0.20 g
Note:
Dulbecco’s PBS in powder form
for tissue culture applications is also
commercially available.
