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Kymograph analysis using Image J

Page history last edited by peiwenchn@gmail.com 6 years, 1 month ago

Image J Kymograph                         by Pei-Wen Chen

  •          To install ImageJ, go to

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)

Kymograph (time space plot) Plugin for ImageJ

 

Author:

J. Rietdorf FMI Basel + A. Seitz EMBL Heidelberg

Date:

2004/04/21
2004/11/11: Macro: some updates / bugfixes
2005/06/07: Macro: bleach correction integrated
2008/10/31: Update of Plugins

Source:

MultipleKymograph_.java,MultipleOverlay_.java,StackDifference_.java,WalkingAverage_.java

Installation:

Download MultipleKymograph_.class, MultipleOverlay_.class, StackDifference_.class, WalkingAverage_.class to the ImageJ plugins folder and restart ImageJ

Download tsp050706.txt (e.g. click with right mouse button and select �save target link as�) to the plugins folder and install it:
ImageJ Menu...Plugins...Macro...Install...

Description:

The kymograph plugin for ImageJ is intended to measure velocities of moving structures in an image time series. Please read the detailed plugin description (including installation intructions)!

 

  •          Start Image J

 

  •          Load a time lapse movie

 

  •          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

 

  •          Analyze>Set Measurements

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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