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Hello everyone
I'm attempting to do something that should be rather easy and yet is proving to be quite troublesome.
Specifically, I am trying to measure the velocities of a particles travelling in a 2D image. For example, a tagged kinesin protein moving along a microtubule track.
I downloaded the multiplekymograph and related plugins, used them as described. I can make nice looking kymographs with this program. I also realize that the kymograph output is not entirely correct, as it the x- and y- axes are pixel values, not distance (x) and time (y) - they need to be converted accordingly. However, when I do the conversions, I get values that I know are not correct. There is NO chance that I have discovered some flaw in published research. The ratios between the values are actually proportionally correct (ie the velocity in range A is 1.5x the velocity in range B), so I think I must be close.
I should also point out that there does appear to be some degree of variation in the exposure time when I'm acquiring the movies, and I'm pretty certain this information is not transferred with the stack from the proprietary acquisition software when incorporated into ImageJ.
Any help/advice would be greatly appreciated!!
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