Testing Cell Printing with Pipette
Prep
- 3% w/v agarose
- 5 ml LB, 150 mg agarose
- heat in 80 deg. bath for 15 min
- 108 ul per mold
Method
- Mold agarose into blocks
- Not using silicon molds, only testing bacterial dispersion on agarose/glass slide.
- waiting 20 min for agarose to dry in PDMS molds, at room temp; in future, will try different drying temps/dessication?
- cover agarose in mold with cover slip
- Place all 6 blocks on rectangular glass slides
- Grew up wild-type MG1655 E. coli to exponential phase, 0.1 ul of solution (measure OD)
- spot two 0.2 ul drops, one on each end of each agarose block/corresp. slide position, wait requisite time for each
Varying:
- wait time
- deposition surface
- agarose: dessicate agarose for 1, 5, 10 min, then add 0.2 ul of cells, then cover slip
- glass coverslip, transfer to agarose: dry cells for 1, 5, 10 min on cover slip, then xfer
Measuring:
Data
| dry time/surface |
1 min |
5 min |
10 min
|
| glass |
x |
x |
x
|
| agarose |
x |
x |
x
|
Fail: Two agarose blocks per slide causes the objective to move away, making focusing impossible. Redo next time with one agarose block per slide.
|
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