Tracking objects under the microscope and characterizing their movements, particularly when studying motors, can be time-intensive. Fortunately, we have several ways to simplify this. One approach involves fitting the Point Spread Function (PSF) with Gaussian, enabling us to accurately determine the object’s center and attain precise nanometer-level accuracy. Or, we can use kymographs.
Kymograph is a great ImageJ plugin that makes life around motors easier.
AuNps-labeled motor(KIF1A) moving along a microtubule
Data ‘cube’ of the movie
Consider the movie from the microscope as a collection of frames within the xy dimension, forming a stack representing the z or time dimension. Ordinarily, our observations are confined to the XY plane, and we study dynamic changes by slicing through the z/time axis. So it is a ‘cube’ of data, with the XY plane on the screen and the z/time axis going into the screen. But what if we look from the bottom/top of the ‘cube’? We can freeze time and see the dynamic in a single frame!
Kymograph!!!
It is fantastic to look at the same thing from a different perspective!
