Difference between revisions of "20220404 cell adhesion"

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(Assembling the chip)
Line 4: Line 4:
 
* Punch holes for fluid inlets
 
* Punch holes for fluid inlets
 
** For 1/16" tubes use OD?? punch
 
** 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  
 
* Place both PDMS and glass slide with clean sides up on glass plate in plasma cleaner  
 
* Follow instructions for [[Plasma treatment]]
 
* Follow instructions for [[Plasma treatment]]

Revision as of 10:48, 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
  • 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

No sterilizing today, everything was done in 420 instead of in 431

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