Difference between revisions of "20220401 cell adhesion"
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=Cell Adhesion= | =Cell Adhesion= | ||
+ | ===Plan for the day=== | ||
+ | [[File:PDMS cast.png|thumb]] | ||
+ | [[File:The valve .png|thumb|The valve]] | ||
+ | *1. Bonding of PDMS together with the microscope slide | ||
+ | *2. Filling the water inside the chambers | ||
+ | *3. Filling fibronectin inside the chambers | ||
+ | *4. Injecting the cells (w/ bubble trap) | ||
− | == | + | ===Bonding=== |
+ | We have ready wafers of our new design showed in the figure to the right. | ||
− | 1. | + | '''Procedure''' |
− | 2. | + | # Cut out the PDMS layer, without cutting the petri dish (make sure there is no dust on the scalpel, use tape) |
− | + | # Once the PDMS layer is removed, make 6 holes with the 1.6 mm punch (make sure the punch is clean before each time you make a hole) | |
− | + | # In the meantime, wash the miscrospoce slide with isopropanol and dry with nitrogen | |
+ | # Place the wafer inside the plasma cleaner for 1 minute, power: 10090 and flow: 60. | ||
+ | # After it is ready, press PDSM onto the miscroscope slide and make sure it is bonded right away, if not something is wrong | ||
+ | |||
+ | === Filling in the water === | ||
+ | |||
+ | '''Procedure (cell lab)''' | ||
+ | # Prepare the valve (see the drawing) | ||
+ | # Prepare 1 mL syringe + a tube with the smallest inner diameter and needles with the size of 20g or 21g | ||
+ | # Connect the tube to the valve | ||
+ | # Fill the syringe with distilled water, connect to one of the chambers on the chip and slowly add the water inside, make sure that the water comes out on the other end of the chamber | ||
+ | # *The chip has disconnected on the first try but we came back with a new chip to continue the procedure | ||
+ | # Prepare fibronectin by taking it out of the freezer and let it out to tin | ||
+ | # *The tube which is hooked up to the valve was too long, we had to cut it in order to connect it to the chip | ||
+ | # Fill the syringe with fibronectin and fill the chamber. | ||
+ | # Place the chip inside a petri dish together with distilled water and place it inside the fridge | ||
+ | '''No sucess, first trial:''' | ||
+ | |||
+ | Once we have added water to the chamber the PDMS layer has disconnected from the microscope slide indicating that there was a mistake when using the plasma cleaner. Possible source of error: unregulated oxygen flow inside the plasma cleaner. | ||
+ | |||
+ | We went back to the other lab to do the procedure for bonding one more time, this time using following parameters flow: 15 time: 2.0 power: 100% pressure: 0.3 mB | ||
+ | |||
+ | Once we had our new chip we went back to the cell lab to continue with the water filling procedure. When filling the chamber with fibronecting, the valve that we have set up earlier did not seem to work; it became difficult filling the chamber with water together with the valve due to the tube being too long which increases the chance of bubbles formation. |
Latest revision as of 14:56, 22 July 2022
Cell Adhesion
Plan for the day
- 1. Bonding of PDMS together with the microscope slide
- 2. Filling the water inside the chambers
- 3. Filling fibronectin inside the chambers
- 4. Injecting the cells (w/ bubble trap)
Bonding
We have ready wafers of our new design showed in the figure to the right.
Procedure
- Cut out the PDMS layer, without cutting the petri dish (make sure there is no dust on the scalpel, use tape)
- Once the PDMS layer is removed, make 6 holes with the 1.6 mm punch (make sure the punch is clean before each time you make a hole)
- In the meantime, wash the miscrospoce slide with isopropanol and dry with nitrogen
- Place the wafer inside the plasma cleaner for 1 minute, power: 10090 and flow: 60.
- After it is ready, press PDSM onto the miscroscope slide and make sure it is bonded right away, if not something is wrong
Filling in the water
Procedure (cell lab)
- Prepare the valve (see the drawing)
- Prepare 1 mL syringe + a tube with the smallest inner diameter and needles with the size of 20g or 21g
- Connect the tube to the valve
- Fill the syringe with distilled water, connect to one of the chambers on the chip and slowly add the water inside, make sure that the water comes out on the other end of the chamber
- *The chip has disconnected on the first try but we came back with a new chip to continue the procedure
- Prepare fibronectin by taking it out of the freezer and let it out to tin
- *The tube which is hooked up to the valve was too long, we had to cut it in order to connect it to the chip
- Fill the syringe with fibronectin and fill the chamber.
- Place the chip inside a petri dish together with distilled water and place it inside the fridge
No sucess, first trial:
Once we have added water to the chamber the PDMS layer has disconnected from the microscope slide indicating that there was a mistake when using the plasma cleaner. Possible source of error: unregulated oxygen flow inside the plasma cleaner.
We went back to the other lab to do the procedure for bonding one more time, this time using following parameters flow: 15 time: 2.0 power: 100% pressure: 0.3 mB
Once we had our new chip we went back to the cell lab to continue with the water filling procedure. When filling the chamber with fibronecting, the valve that we have set up earlier did not seem to work; it became difficult filling the chamber with water together with the valve due to the tube being too long which increases the chance of bubbles formation.