Revision as of 10:41, 4 April 2022

Followed the procedure

Assembling the chip

• Clean glass slide with IP and N2
• Punch holes for fluid inlets
  • For 1/16" tubes use OD?? punch
  • For thin teflon tube use OD?? punch
• Place both PDMS and glass slide with clean sides up on glass plate in plasma cleaner
• Follow instructions for Plasma treatment
• Placed the PDMS on top of the glass slide and watch it attach
• Some pressure needed to be applied to bond

Filling the chip

For sterilizing and filling the chip with water and fibronectin you need

• 1ml syringe with
• 20G needle and
• 2cm long 1/16" teflon tube on the needle.
• small beaker
• ethanol
• distilled water
• Eppendorf tube of fibronectin
• Cells in fresh media
• Inlet and outlet tubes
  • Measure lengths of inlet and outlet tubes and cut
  • Sterilize the tubes

Sterilizing

• If you want a sterile chip move each object into LAF bench once they have been sterilized
• Spray chip with ethanol and rub off glass side
• Assemble syringe, needle and teflon tube
• Spray ethanol and rub outside
• Rinse beaker in ethanol and pour in a small amount of ethanol
• Retract syringe half way in air, and fill teflon tube with ethanol
• Empty syringe of ethanol and air, make sure it is dry
• Empty beaker of ethanol

Water filling

• Fill a small amount of distilled water in beaker
• Retract <1ml distilled water into tube/needle/syringe
• Fit in inlet of chip and fill slowly
• Empty syringe and needle to be ready to use for fibronectin

Fibronectin filling

• Take one Eppendorf with 0.5 ml fibronectin form freezer
• Thaw the fibronectin
• Retract fibronectin into tube/needle/syringe
• fit in inlet of chip and fill slowly
• Attach inlet and outlet tubes
• Incubate for 30 min in incubator
• Rinse with PBS
  • Details?
  • Pour ~?? ml PBS in sterile beaker
  • Fill sterile ?? ml syringe+needle with PBS
  • Slowly flow PBS through chip

Cell filling