Difference between revisions of "20211806 microfluidics"
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=== Collagen coating: === | === Collagen coating: === | ||
| − | We coated Annikens chip (100 um) with collagen. We injected the collagen by using a 5 mL syringe and with a flow of around 50 uL/min. Stopped the flow afterwards and let the collagen settle for one hour. Aspirated the chip by using a 5 mL syringe multiple times. | + | We coated Annikens chip (100 um) with collagen. We injected the collagen by using a 5 mL syringe and with a flow of around 50 uL/min. Stopped the flow afterwards and let the collagen settle for one hour. Aspirated the chip by using a 5 mL syringe multiple times. Then we used Elveflow pressure controller OB1 with nitrogen gas for about 30 minutes to dry the chip completely. |
== Casting PDMS == | == Casting PDMS == | ||
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== Passaging == | == Passaging == | ||
We passaged the cells as usual. For detail see [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/Cell_culturing here]. We kept one flask and changed one flask. We took out 2 ml from the old flask into a 3 ml syringe. | We passaged the cells as usual. For detail see [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/Cell_culturing here]. We kept one flask and changed one flask. We took out 2 ml from the old flask into a 3 ml syringe. | ||
| + | |||
| + | == Injecting cells into chip and preparing for timelaps == | ||
| + | We used tubes marked "For Elveflow material" all day. We used hypodermic needle in size 0.60 all day (also for the collagen coating). | ||
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| + | We filled the tube separately by using a 3 mL syringe manually with PBS and then used the same syringe to clean the chip with PBS (automatically). | ||
| + | |||
| + | We changed the syringe to a 3 mL syringe with cells in media. Kept the same tube. Injected the cells. | ||
| + | |||
| + | We changed the syringe to a 5 mL syringe with CO2 ind. media. Kept the same tube. Miscalculated and set the flow rate at 1 mL/min instead of 1 uL/min. Blew all the cells out. | ||
Revision as of 16:26, 18 June 2021
Contents
Preparing collagen and coating chip with collagen
Collagen preparation:
We did a 1:1000 dilution of acetic acid (17.5 M) - we used 10 uL of acetic acid and 10 mL of sterile water.
We did a 2:3 dilution of collagen - we used 200 uL of 5 mg/mL collagen and 300 uL of the diluted acetic acid solution.
Collagen coating:
We coated Annikens chip (100 um) with collagen. We injected the collagen by using a 5 mL syringe and with a flow of around 50 uL/min. Stopped the flow afterwards and let the collagen settle for one hour. Aspirated the chip by using a 5 mL syringe multiple times. Then we used Elveflow pressure controller OB1 with nitrogen gas for about 30 minutes to dry the chip completely.
Casting PDMS
We casted two new chips - Annikens (100 um) and Thomas' (25 um).
Passaging
We passaged the cells as usual. For detail see here. We kept one flask and changed one flask. We took out 2 ml from the old flask into a 3 ml syringe.
Injecting cells into chip and preparing for timelaps
We used tubes marked "For Elveflow material" all day. We used hypodermic needle in size 0.60 all day (also for the collagen coating).
We filled the tube separately by using a 3 mL syringe manually with PBS and then used the same syringe to clean the chip with PBS (automatically).
We changed the syringe to a 3 mL syringe with cells in media. Kept the same tube. Injected the cells.
We changed the syringe to a 5 mL syringe with CO2 ind. media. Kept the same tube. Miscalculated and set the flow rate at 1 mL/min instead of 1 uL/min. Blew all the cells out.
