Particle analyzing for low contrast image

test10657.tif (3.0 MB)

Hi, I got a microscope image like this and try to figure out how to do particle analysis with it. The challenge is the contrast is low so I can’t really get a good binary image to show where circles are.

Any idea?


The ImageJ forum became a helpful archive.

This solution might help you with your analysis:

For other solutions search this forum for droplets, circular objects and rings.

1 Like

Welcome to the Forum, Zhong Junjie. You have a broad distribution of what appear to be spheroidal particles. A coverslip can flatten thicker bubbles or soft particles. Your test image is also saturated (take a look at the histogram below). Note the clipping of the right side from the bright large particles (are these particles or bubbles?).


At the other end of the size distribution you have some very small particles.

One problem with digital image analysis is that you always get an answer. If one has not measured all the particles in the distribution with sufficient precision, your answer will be biased. With broad size distributions like this, one needs to measure LOTS of particles to get a meaningful size distribution.

Some of these concepts are nicely illustrated in the Image Processing for Life Sciences course on EdX.

A key paper you should look at for particle analysis is: H.Masuda and K. Iinoya, “Theoretical study of the scatter of experimental data due to particle size distribution,” J. Chem Eng. Japan, 4, 60-66 (1971).

Best regards,
John Minter

Thanks, I will try and see!

Thank you! I will check that paper.