Touching particle accurate counting

Posted by Al Menia on Jan 14, 2015; 5:02pm
URL: http://imagej.273.s1.nabble.com/Touching-particle-accurate-counting-tp5011214.html

Hi,

I am desperately trying to find an automatic way to count diamonds (approx 100 um size) on a metallic substrate on SEM imaging. The problem is that they are sometimes overlapping, making them hard to separate, even by using the watersheding tool. (you'll see very big parts not affected by this tool)
The method I use is not very accurate:
- gaussian blur or substrack background (sometimes it works better)
- tresholding
- watersheding
- analyse particle (area 200-infinite, circularity 0.35-1.00)

There are around 620 diamonds in the dense onem and 240 in the less dense one. I can only manage to reach a 10% accuracy in the counting, but I would like to reach 1%.

Does anybody knows a better way to do this, or knows a good plugin for this kind of work?
A good thing would be to be able to force the watersheding above a given area, so that I don't have big agglomerates, but I don't know if this already exists...

Thanks,