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

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 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 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.'