User:Daniel Goodman/Notebook/Cluzel/2010/02/22

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Cluzel Lab Notebook
Daniel Goodman
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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
  • 6 agarose block

Method

  1. Mold agarose into blocks
    1. Not using silicon molds, only testing bacterial dispersion on agarose/glass slide.
    2. waiting 20 min for agarose to dry in PDMS molds, at room temp; in future, will try different drying temps/dessication?
    3. cover agarose in mold with cover slip
  2. Place all 6 blocks on rectangular glass slides
  1. Grew up wild-type MG1655 E. coli to exponential phase, 0.1 ul of solution (measure OD)
  2. 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
    • 1 min, 5 min, 10 min
  • 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:

  • spot dispersion diameter

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.