Sortostat/Experiments/Two reactor chip

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This is a summary of successful experiments done with the new version of the chip (two reactors).

Feb2006

  • What was the date of this experiment
  • post image

4/21/07

  • post images of %CFP and cell counts

5/29/07

  • Media:
    • 0.4% glycerol
    • Amp
    • BSA(2mg/ml)
  • Temp: 37C (set to 40.4 on the incubator).
    • What was the offset?
  • Fluid: 5.1 psi
  • Push down: 21.9 psi
  • Push up: 20.2 psi
  • Clean period:15.9 min
    1. of sorts / clean: 10
  • Number of screens / sort: 2
  • Cleaning block length (min): 1
  • Pump cycle period (ms): 400
  • Ratio history length: 15
  • Pump period between screens (sec): 5
  • Wash sorting chamber period (ms): 1000
  • Media inlet source: center
  • Feeder fill period (ms): 1000
  • Clean period (ms): 15000
  • Pressurize period (ms): 1000

Refocus

  • 4465, 4756, 6863, 8375, 9924, 10643, 10814, 11581, 12265, 13087

Flips

  • 4888
  • 6864 - against YFP
  • 9068 - against CFP
  • 10324 - against YFP
  • 11586 - random sorting
  • 13087 - against CFP

7/11/07

7/11/07

  • cleaning period = 15.9
  • same as 5/29/07, otherwise

7/11/07b

fluid 5.5 pushup: 19.64 pushdown: 21.80

  • cleaning period = 14.8
  • Focus: 1375, 1984, 2102, 3648, 4068, 5600, 6022, 7502, 8008, 9100, 9718, 9927, 11062, 13036, 13638, 13662, 15358
  • 1436 - Started sorting againstYFP
  • 9927 - turned off sorting
  • 13708 - started sorting againstCFP

9/11/07

  • 40 to 60% zig-zag experiment.
    • If possible do pulse selection as well
  • cleaning period = 15.9
  • same as 5/29/07, otherwise
  • Focus and flipped the 2x magnification at #63 (I was seeing cells outside the sorting chamber).

12/2/07

  • 1418 - focused
  • 3832 - focused for BF images.

12/18/07

  • 64 - switched to sorting against CFP, since it looked like they might be growing faster (though too early to tell, really)
  • 94 - turned off directional sorting
  • 748 - focused
  • 752 - directional sorting against CFP
  • 1321 - focused
  • 3228 - got through >3000 w/o a memory leak crash, as a point of reference. (though PF at 1.2MB)
  • Also, note that this chip has been in constant use since 12/2 (16days), although i did have to reload the pump valves at day 14, b/c one had dried out. I think its reasonable that we could keep a culture going in here for at least 16 days (though in this run, i swapped n new cells a couple times, though one set was in there for 10 days) - and probably for much longer if you periodically reloaded the pump valves.

12/20/07

  • 462 - directional sorting against CFP

12/22/07

  • Using C/Y (3k3)
  • Mixed CFP (OD = 0.12) and YFP (OD=0.10), so they would be present 50/50 from cultures that had been inoculated from a colony ~6 hours earlier.
  • 112 - turned on sorting (non-directional)
  • 817 focused

12/23/07

  • 200 - turned on sorting (non-directional)
  • 230 - focused
  • 494 - focused and turned on directed sorting against CFP, also turned on the "flipping" from 0.4-0.6

12/24/07

  • 20.5 cleaning period, which should be a doubling time of 0.9hr.
  • 368 - set to against YFP / focused
  • 492 - actually turned on the flipping 0.4-0.6 switch. So should flip to against CFP very soon (if not immediately).
  • 1224 - focused
  • 1700-focused

12/25/07

  • 19.3 cleaning period, which should be a 0.85 hr doubling time.
  • 634 focused
  • Good example of non-directional w/ sorting, however the counts for the CFP were consistently 4-5 cell high, so I changed the std_mult variable to be 5 for the 12/26 run.

12/26/07

  • 19.3 cleaning period, which should be a 0.85 hr doubling time.
  • 166 - focused
  • 564- changed to sorting against YFP (was sorting against CFP for a fair number of sorts before this)

12/28/07

  • 715 - turned on non-directional sorting
  • 720 - turned on directional sorting (in this case that just means selecting for GFP cells)
  • 1630 - focused
  • 3536 - focused & turned off directional sorting
  • 4067 - focused

1/15/08

  • push down, push up = 20psi
  • fluid = 5.6 psi
  • 1 - started with cleaning only
  • 155 - focused
  • 162 - turned on sorting (directional) in favor of GFP
  • 462 - focused (~16 total cells, 4 of which were glowing)
  • 524 - switched sorting to non-directional (there was a bug so didn't really)
  • 708 - fixed the bug in the sorting, in the process I did two cleaning events in quick succession, but I restarted at the same cleaning event it was at previously.
  • 1560 - changed sorting to directional in favor of GFP
  • 1950 - focused
  • 2550 - focused
  • 4136 - focused
  • 4716 - focused
  • 5318 - focused
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