Difference between revisions of "20210325 3Dcell labnotes"
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Dagkd@uio.no (talk | contribs) (Created page with "== Plan for the day == === Equipment === * 0.5 ml Eppendorf tubes * tube rack for holding Eppendorfs upright * pipette * distilled water (or growth medium) * low gelling temp...") |
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* for 0.5 ml 1 wt % agarose gel | * for 0.5 ml 1 wt % agarose gel | ||
* 0.5 g = 500 mg water = 500 ul | * 0.5 g = 500 mg water = 500 ul | ||
| − | * weigh 5 mg agarose | + | * weigh 5 mg agarose |
* put eppendorf in water bath and mix with pipette while heating | * put eppendorf in water bath and mix with pipette while heating | ||
| Line 31: | Line 31: | ||
** adjust counter pressure with outlet syringe to get lamella filled | ** adjust counter pressure with outlet syringe to get lamella filled | ||
** when filled, aspire from inlet syringe and press with outlet syringe to get an open channel between lamellae | ** when filled, aspire from inlet syringe and press with outlet syringe to get an open channel between lamellae | ||
| + | |||
| + | == Execution == | ||
| + | |||
| + | * weighed 6 mg = 0.006 g + 0.6 g water = .6 ml = 600 ul water pipetted into 1.5 ml Eppendorf | ||
| + | * put eppendorf in water bath on hot plate (alu paper as lid and support for eppendorf) | ||
| + | * cut sharp tip off an 18 gauge syringe tip | ||
| + | * blunted (320 gauge paper) the tip | ||
| + | * curved the tip to get access under condenser of microscope | ||
| + | * 1st try: largest | ||
Revision as of 13:00, 25 March 2021
Contents
Plan for the day
Equipment
- 0.5 ml Eppendorf tubes
- tube rack for holding Eppendorfs upright
- pipette
- distilled water (or growth medium)
- low gelling temperature agarose
- small beaker
- heater
- precision balance
- microscope at 37C
Gel procedure
- start microscope heater to 37C
- put circuit and pipettes inside microscope box to heat to 37 C
- heat water bath in small beaker to 60C on hot plate
- for 0.5 ml 1 wt % agarose gel
- 0.5 g = 500 mg water = 500 ul
- weigh 5 mg agarose
- put eppendorf in water bath and mix with pipette while heating
Gel into microfluidic procedure
- Start PC and camera on microscope, choose magnification and test imaging of circuit
- connect 2 ml syringe to output of gel channel
- prepare connection of ul micropipette to input of channel
- withdraw tube/syringe tip from circuit inlet
- Fill ul pipette with agarose gel solution
- reconnect ul pipette to gel channel input
- On microscope with camera recording
- inject gel
- adjust counter pressure with outlet syringe to get lamella filled
- when filled, aspire from inlet syringe and press with outlet syringe to get an open channel between lamellae
Execution
- weighed 6 mg = 0.006 g + 0.6 g water = .6 ml = 600 ul water pipetted into 1.5 ml Eppendorf
- put eppendorf in water bath on hot plate (alu paper as lid and support for eppendorf)
- cut sharp tip off an 18 gauge syringe tip
- blunted (320 gauge paper) the tip
- curved the tip to get access under condenser of microscope
- 1st try: largest
