Difference between revisions of "20220404 cell adhesion"
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* Clean glass slide with IP and N2 | * Clean glass slide with IP and N2 | ||
* Punch holes for fluid inlets | * Punch holes for fluid inlets | ||
| − | ** | + | ** Used 1/16" punch to create the holes |
| − | |||
* 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]] | ||
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** Sterilize the tubes | ** Sterilize the tubes | ||
== Sterilizing == | == Sterilizing == | ||
| − | + | No sterilizing today, everything was done in 420 instead of in 431 | |
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
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== Water filling == | == Water filling == | ||
* Fill a small amount of distilled water in beaker | * Fill a small amount of distilled water in beaker | ||
* Retract <1ml distilled water into tube/needle/syringe | * Retract <1ml distilled water into tube/needle/syringe | ||
* Fit in inlet of chip and fill slowly | * Fit in inlet of chip and fill slowly | ||
| + | ** Needle into tube first, then tube into inlet | ||
* Empty syringe and needle to be ready to use for fibronectin | * Empty syringe and needle to be ready to use for fibronectin | ||
| + | |||
== Fibronectin filling == | == Fibronectin filling == | ||
* Take one Eppendorf with 0.5 ml fibronectin form freezer | * Take one Eppendorf with 0.5 ml fibronectin form freezer | ||
| Line 50: | Line 44: | ||
** Slowly flow PBS through chip | ** Slowly flow PBS through chip | ||
== Cell filling == | == Cell filling == | ||
| + | [[File:Cell Colony April 4th.jpg|thumb|Cell Colony April 4th]] | ||
| + | [[File:Cell adhesion .png|thumb|Cell adhesion first 10 minutes]] | ||
Latest revision as of 13:51, 4 April 2022
Followed the procedure
Contents
Assembling the chip
- Clean glass slide with IP and N2
- Punch holes for fluid inlets
- Used 1/16" punch to create the holes
- 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
- Needle into tube first, then tube into inlet
- 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
