- Took a colony from fridge plate of MG1655 w/ GFP, put in 4 mL LB with 4 uL amp in 37°C shaker. (~10:30 AM)
- Made 5 mL of 5% agarose, and 5 mL of 3% agarose. (~10:40 AM)
Make chip impression in solidified gel
- Try quick experiment based on results from Wednesday; can we make features on a gel by pressing down?
Results: no features found with significant 'push' on 3% gel.
Try transferring again (w/ features)
- Scope access at 4PM
- Start gel pouring at 2:30 - Poured 3% at 2:30, poured 5% at 2:50
- Not enough gel poured in mold, features did not form (but still continued w/ experiment)
- In future, try to use more than 108, maybe 110. Removing bubbles (which happens often) often reduces height of agarose blob in mold and ruins features.
- 3% - couldn't find any cells, this could be because surface was concave so cells did not touch agarose
- 5% - cells found. Large (several screens), toward bottom of mold, so hard to tell extent - but clear boundaries were apparent!
- Images saved in dbg, under 3-5-(5pct-gfp/phase)*
- Used blade on handle to pry gel from out of mold so mold sat on surface of blade
- Then used tweezers to grab long sides of mold from off of cell-coverslip, moved to slide to be imaged
- Try spotting smaller amount; maybe using a needle, just a pipette tip (without taking up liquid) or something similar (nanodrop?).
- Make journals for metamorph so multiple screen can be taken automatically
- Figure out how stage increments in metamorph maps to nm/μm
- Saved some slide positions for the blob in my blue notebook, maybe plot them clumsily later when we figure out slide position mapping
- Play with a few featureless gels, practice flipping & transferring