Sortostat/Notebook: Difference between revisions
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==To Do List== | |||
*Figure out the MATLAB memory leak | |||
**I suspect this is because I'm not closing the image files properly in the MATLAB script that runs, or it could be something more annoying. | |||
**Does the leak influence the timing of the events, by making the image processing take too long? (e.g. why does the timing graph get noisy after awhile?) | |||
**Is it the MATLAB thread that blows up or the LabView? it's MATLAB. | |||
**[http://www.mathworks.com/support/tech-notes/1100/1106.html MATLAB help page here might be useful.] | |||
*output the direction of sorting to the output file | |||
*Clean up the Image Processing MATLAB scripts | |||
==Done== | |||
*Why do I see any variability in the timing? Timing between cleaning events is rock solid, the timing between sorts is influenced by the time it takes to process the image. | |||
*Change the code so that when flipping based on the CFP it is the moving average that is used, rather than a single image. | |||
*Make the list of experiments to evaluate the device performance limits. | |||
**Sorting at various dilution rates, followed by a sort to extinction. | |||
==Experiments== | ==Experiments== | ||
*[[Sortostat/Experiments/Two reactor chip]] | *[[Sortostat/Experiments/Two reactor chip]] | ||
*[[Sortostat/Troubleshooting]] | |||
===Future Experiments=== | ===Future Experiments=== | ||
'''replace bulb before these experiments''' | '''replace bulb before these experiments''' | ||
*check on pH of the media if we are adding AHL, it needs to be <8 for AHL to be stable. | |||
*Zig-zag run where we vary from 0.4 to 0.6 so that we can get more sampling of the selective pressure. | *Zig-zag run where we vary from 0.4 to 0.6 so that we can get more sampling of the selective pressure. | ||
*Maintenance of a slower growing variant vs. a faster growing "mutant". | *Maintenance of a slower growing variant vs. a faster growing "mutant". | ||
**Could demonstrate this with a slower growing CFP vs YFP or via something like F2620. | **Could demonstrate this with a slower growing CFP vs YFP or via something like F2620. | ||
*Zig-zag experiment with different dilution rate (e.g. different total number of cells). | *Zig-zag experiment with different dilution rate (e.g. different total number of cells). | ||
*Get the exact size of sorting chamber by cell counting | |||
*F2620 is in MG1655 whose flagella might cause a problem | |||
**Need to co-transform with RFP expression plasmid anyway. | |||
*We already have the zig-zag automated, but should include a "pulse" as well (e.g. /-\_/-\) where it has regions with no selection. | |||
==Modeling== | ==Modeling== | ||
Line 15: | Line 35: | ||
**No, looked at this before last lab meeting, but should write it up. | **No, looked at this before last lab meeting, but should write it up. | ||
*Do fluctuations in the total cell count correlate with anything? | *Do fluctuations in the total cell count correlate with anything? | ||
*2/18/06 lab notebook page has some information about the appropriate sorting volume to use in the model, move this to the wiki | |||
===Output=== | ===Output=== | ||
Line 21: | Line 42: | ||
*Sortostat Modeling Stochastic Sim doc - description of the stochastic simulation of Sortostat performance. | *Sortostat Modeling Stochastic Sim doc - description of the stochastic simulation of Sortostat performance. | ||
== | ===Analytical model=== | ||
* | *put the cleaning period / dilution rate formula derivations on the wiki. | ||
*write up the chemostat equations as they relate to the SS cell concentration (e.g. limiting substrate? Should we reduce the concentration of glycerol?) | |||
* | *These come from the SortostatTiming.m matlab model: | ||
** | **12.5 min between cleaning (doubling time = 0.55) | ||
* | **13.6 min between cleaning (doubling time = 0.60) | ||
* | **14.8 min between cleaning (doubling time = 0.65) | ||
**15.9 min between cleaning (doubling time = 0.7) | |||
**17min between cleaning (doubling time = 0.75) | |||
**18.2min between cleaning (doubling time = 0.8) | |||
**19.3min between cleaning (doubling time = 0.85hr) | |||
**20.5min between cleaning (doubling time = 0.9hr) | |||
**21.6min between cleaning (doubling time = 0.95hr) | |||
**22.7min between cleaning (doubling time = 1hr) | |||
**23.9min between cleaning (doubling time = 1.5hr) | |||
==LabView== | |||
*why do we need to initialize the load paraemters array? | |||
==Methods== | |||
*[[Sortostat/Methods/Receiving chips]] | |||
==References== | ==References== | ||
*[[Sortostat/Foundry report card]] - post notes on each chip fabricated so can document common failure points in fabrication to better avoid them. | *[[Sortostat/Foundry report card]] - post notes on each chip fabricated so can document common failure points in fabrication to better avoid them. |
Latest revision as of 09:20, 23 December 2007
To Do List
- Figure out the MATLAB memory leak
- I suspect this is because I'm not closing the image files properly in the MATLAB script that runs, or it could be something more annoying.
- Does the leak influence the timing of the events, by making the image processing take too long? (e.g. why does the timing graph get noisy after awhile?)
- Is it the MATLAB thread that blows up or the LabView? it's MATLAB.
- MATLAB help page here might be useful.
- output the direction of sorting to the output file
- Clean up the Image Processing MATLAB scripts
Done
- Why do I see any variability in the timing? Timing between cleaning events is rock solid, the timing between sorts is influenced by the time it takes to process the image.
- Change the code so that when flipping based on the CFP it is the moving average that is used, rather than a single image.
- Make the list of experiments to evaluate the device performance limits.
- Sorting at various dilution rates, followed by a sort to extinction.
Experiments
Future Experiments
replace bulb before these experiments
- check on pH of the media if we are adding AHL, it needs to be <8 for AHL to be stable.
- Zig-zag run where we vary from 0.4 to 0.6 so that we can get more sampling of the selective pressure.
- Maintenance of a slower growing variant vs. a faster growing "mutant".
- Could demonstrate this with a slower growing CFP vs YFP or via something like F2620.
- Zig-zag experiment with different dilution rate (e.g. different total number of cells).
- Get the exact size of sorting chamber by cell counting
- F2620 is in MG1655 whose flagella might cause a problem
- Need to co-transform with RFP expression plasmid anyway.
- We already have the zig-zag automated, but should include a "pulse" as well (e.g. /-\_/-\) where it has regions with no selection.
Modeling
- Sortostat/Model
- How much faster can we drive it given the timing?
- Are there more or less cells in a sort vs a screen?
- No, looked at this before last lab meeting, but should write it up.
- Do fluctuations in the total cell count correlate with anything?
- 2/18/06 lab notebook page has some information about the appropriate sorting volume to use in the model, move this to the wiki
Output
- Sortostat mFiles doc - list of all the mfiles that matter (except the image processing). These are all commented and cleaned up at this point.
- Sortostat Modeling Probability Distributions doc - description of the modeling of the probability distributions for the number of cells in the sorting chamber and how many are CFP or YFP.
- Sortostat Modeling Stochastic Sim doc - description of the stochastic simulation of Sortostat performance.
Analytical model
- put the cleaning period / dilution rate formula derivations on the wiki.
- write up the chemostat equations as they relate to the SS cell concentration (e.g. limiting substrate? Should we reduce the concentration of glycerol?)
- These come from the SortostatTiming.m matlab model:
- 12.5 min between cleaning (doubling time = 0.55)
- 13.6 min between cleaning (doubling time = 0.60)
- 14.8 min between cleaning (doubling time = 0.65)
- 15.9 min between cleaning (doubling time = 0.7)
- 17min between cleaning (doubling time = 0.75)
- 18.2min between cleaning (doubling time = 0.8)
- 19.3min between cleaning (doubling time = 0.85hr)
- 20.5min between cleaning (doubling time = 0.9hr)
- 21.6min between cleaning (doubling time = 0.95hr)
- 22.7min between cleaning (doubling time = 1hr)
- 23.9min between cleaning (doubling time = 1.5hr)
LabView
- why do we need to initialize the load paraemters array?
Methods
References
- Sortostat/Foundry report card - post notes on each chip fabricated so can document common failure points in fabrication to better avoid them.