User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/02/04

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Whole Casein data

So I went ahead and tracked some MTs in whole casein. I've ordered the data such that each FOV has the MTs velocities increasing. Each FOV is approximately 2 minutes worth of imaging.

I think that this data does show that there is either a huge spread in velocities early on when you make a slide or that there is groups of MTs moving at different velocities.

I am going to have to think about this more...

Steve Koch 19:46, 4 February 2010 (EST): Very cool! I too do not understand. It does seem like slow species disappear over first couple minutes. Then wide "medium" range persists until 12 minutes. Whereas "fast" species doesn't emerge until 10 minutes. I agree that distinct groups are not crystal clear. Within a given data set, this would show up as a plateau, which shows in the 6 minutes data. If all data were combined, there appears to be a cluster around 750. But there are also all kinds of other speeds, which calls for a different explanation than just # of protofilaments.
Another mystery is the disappearance of "slow" species and the emergence of "fast" species. I don't get this at all, unless MTs are annealing in some way? Have you seen any evidence yet of an individual MT accelerating?
One fortunate thing (perhaps) is that the spread decreases at 12 minutes and greater. Indicating that 15 minutes (or higher) may be a good time to use for comparison
Could this have anything to do with antifade? Or other components of the system?
Also, you may want to put this figure in a mindmap for your paper / poster (Larry knows how to do this)
  • Andy Maloney 21:19, 4 February 2010 (EST): I have no idea about the antifade. Yes, I have seen a MT accelerate quite dramatically.
    • Steve Koch 22:13, 4 February 2010 (EST): Make a note of that MT if you can! As for antifade, since it's enzymatic, it could have something to do with this time dependence. No hypothesis for how, though. I still lean towards some property of the MTs ... Down the road, you could certainly do a study where you add certain components to the motility assay after different times. E.g., mix up the antifade, then add MTs an hour later, etc. Not a high priority yet, though.