Difference between revisions of "20210325 3Dcell labnotes"

Revision as of 13:52, 25 March 2021

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

Analysis

• FOV is 3 x 2.4 mm, see image scale.png