Image J Kymograph by Pei-Wen Chen
http://rsb.info.nih.gov/ij/index.html
- To install related plugins (you only need "MultipleKymograph.class" for this analysis), go to
http://www.embl.de/eamnet/html/body_kymograph.html
and follow instructions on the webpage (below)
- Image>Stacks>Z Projection
Choose the start and stop slice, usually the first and the last time point of the movie
Projection Type:choose “Max Intensity”
- Save the Z projection image as a new Tiff file
- Line tool>right click & choose “Segmented Line”
- Mark the track of interest on the Z projection image
- Click on the original time lapse movie file
- Edit>Selection>Restore Selection
You should see a line appear on the time lapse movie file
- Plugins>MultipleKymograph
Line width: choose 5
- Save the Kymograph image as a new Tiff file
The Y axis is time and the unit is the time intervals of the movie
The X axis is the distance the object of interest travels along the chosen segmented line and the unit is the pixel size of the movie
Choose “Bounding rectangle”
- Line tool>right click & choose “Straight Line”
- Trace the track of interest
- Analyze>Measure (or Ctrl+M)
- Use “Width” and “Height” to calculate velocity
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