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CELL COUNTING USING IMAGEJ

Particle counting can be done automatically using ImageJ, a free software for
image processing and analysis in Java.
In a few and very simple steps you can count the cells in your biological
experimental image.
Automatic particle analysis requires the image to be a binary image i.e. black or
white. The software needs to know exactly where the edges are to perform
morphology measurements. A threshold range is set and pixels in the image
whose value lies in this range are converted to black; pixels with values outside
this range are converted to white (or vice-versa depending on what the user
requests).
The image can be converted to a binary image via the menu command
Image/Type/8-bit.
In a few and very simple steps you can count the cells in your biological
experimental image.

Download WCIF IMAGEJ at link


http://www.uhnresearch.ca/facilities/wcif/imagej/installing_imagej.htm

Install WCIF IMAGEJ following the website instructions

Open WCIF IMAGEJ

Import Image File. The menu command File/Open will import common file
formats(e.g. JPEG, GIF, BMP, PGM, PNG, TIFF)

Convert image into a binary image from Image/Type/8-bit

Click on Plugins/Particle Analysis/ Nucleus counter, a window opens up

Enter the size range which should be counted

Opt to perform a background correction. We suggest to select this option.

Select the automatic thresholding method. This can be either "Current",


"Otsu", Maximum Entropy", "Mixture Modeling" or k-means" clustering. We
suggest k-means clustering which can segment an image in to multiple
bands.

Opt to perform a "Smooth" filter. We suggest mean 3x3

Opt to perform a watershed separation. We suggest to select this option.

Opt to add the particles to Region Of Interest (ROI) manager.

Opt for a summary.

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