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| | = [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/PDMS PDMS casting and bonding] = | | = [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/PDMS PDMS casting and bonding] = |
| | = [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/Microfluidics Microfluidic chip production SOPs] = | | = [https://wiki.uio.no/mn/fysikk/laglivlab/index.php/Microfluidics Microfluidic chip production SOPs] = |
| − | = Fibronectin patterning = | + | = [Surface patterning Surface patterning] = |
| − | == First protocol ==
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| − | Tested 06.02.2023
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| − | Equipment and materials
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| − | * Fibronectin (eppendorfs in freezer 1 mg/ml), dilute to 50 ng/ml (? check)
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| − | * Fluorescent fibronectin added in ration 1/10
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| − | * Matec 35mm petri dishes with glass bottom plasma cleaned (no O2, 100% 5 min). For future: test cleaning and plasma protocol for activation. Petri dishes moved to 431 before stamping. Too long time?
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| − | * Plasma cleaning: handheld could not be used because too large distance to glass bottom
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| − | * PDMS stamps: cut out from top of patterned silicon wafer. The wafer was broken and maybe some fragments were transferred. Check in WLI. Some messing around because they were turned around. checked in microscope. They are hydrophobic.
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| − | * Fibronectin solution drops (about 100 ul) on each stamp. The drops sit on top of the stamp. Pipette tip used to "drag" drop out to the edges. Drops left on stamp for 1h in laf bench under aluminum foil to prevent bleaching.
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| − | * Remove drop from stamps with pipette + nitrogen flow
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| − | * wash with sterile water (not distilled) and remove with N2 flow (other protocols: PBS)
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| − | * Apply stamp to glass, leave for 10 minutes
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| − | * Remove stamp (we didn't, but decided it would be best to hold the petri dish up side down to remove the stamp quickly without touching the glass surface again)
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| − | * Add BSA solution (powder mixed into PBS 5mg/ml, filter). Leave for 1 h.
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| − | * Clean 3 times with PBS, remove with pipette
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| − | * Add cells suspended in media.
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| − | * We left it only for 10 minutes before removing media with suspended cells and adding new media. The idea was to not have too many cells that did not find patterned areas.
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