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Mammalian Cell Freezing Media Recipe
90% FBS
10% DMSO
Construction of PDMS - Glass Culture Platform
*PDMS Construct*
1. Heat the oven to 65oC
2. Poly(dimethyl siloxane) (PDMS) will be used in a 10:1 ratio of base to
curing agent (SYLGARD 184 Silicone Elastomer)
3. In a plastic cup, weigh approximately 30g of the elastomer base.
4. Add 3g of the curing agent.
5. Mix vigorously
6. Pour the PDMS mixture on top of mold
7. Place in a vacuum desiccator and continue to de-gas until all bubbles
have been expelled from the PDMS
8. Cure the PDMS at 65oC for 1 hour
9. Cut the construct out using a razor blade or scalpel
10. Bore holes of the appropriate size for tube placement
*Plasma Treatment*
1. Clean the PDMS and glass thoroughly using isopropanol, methanol,
acetone, and ethanol.
2. Dry with air
3. Press the pump and then the ventilation button to open the door of the
O2 plasma machine
4. Place the PDMS channel side up and the glass clean side up
5. Close the machine door
6. Press the ventilation and then the pump button to seal the door
7. Press the gas button to allow O2 to saturate the space
8. Wait 1-2 minutes until the indicator reads 10
9. Press the generator button to plasma treat the glass and PDMS for 10s.
This creates a hydrophilic surface for the irreversible bonding of PDMS to
glass.
10. Press the pump and then the ventilation button to remove the
materials
11. Press the PDMS to the glass to seal the construct
12. Place in 65oC for 10 minutes
*Tube Hole Reinforcement (optional)*
1. Either save some uncured PDMS from the previous procedure or make a
small amount while the construct is in the oven after plasma treatment
2. Place small sections of tubing into each bored hole
3. Paint a small amount of PDMS around the tubing sufficient to reinforce
any tearing that may have occurred using a pipette tip
4. Cure at 65oC for approximately 1 hour
