Open ImageJ
File à Open à cell count.jpg
Click “開啟(O)”
Edit à Options à Conversions
Check “Scale When Converting”
Click “OK”
Image à Type à 16-bit
Image à Adjust à Threshold
Adjust threshold until you are satisfied
Click “Apply”
* To fill the hole in cells
Process à Binary à Fill Holes
* To separate two overlapping cells,
Process à Binary à Watershed
Analyze à Analyze Particles
Size (pixel^2): to count all particles, 0 – Infinity
Circularity: 0.00: a straight line; 1.00: a perfect circle;
Check cell size and circularity
Click “OK”
Cell number is 68 at current setting.
However, check Areas in Results windows, some noises are included. So, change area size
Analyze à Analyze Particles
Show: outlines
Size (pixel^2): to count all particles, 6 – Infinity
Click “OK”
Show: mask overlay
Cell number from 68 à 64
