I performed a particle analysis using images (16bit) taken with a confocal microscope. Before I performed the particle count, I had to convert the 16 bit image to an 8 bit image. Why is this step necessary? Is it just “easier” to calculate because of the reduced number of colors?
Thanks for helping
Hello and welcome Rike,
The reason is because the macro you are trying to use only works with 8 bit binary images. in other words a Range of 0-255 vs 0-65536.
I guess, as @smith_robertj already indicated that you thresholded your image and thus created a binary image (only black and white, which are the values 0 and 255, respectively). Actually, a 1-bit image would be sufficient, but standard wise 8-bit images are taken for this task (at least in ImageJ).
The indication binary tells you that it is practically an on/off system. This is necessary to make plugins like the
Analyze Particles... recognize pixels assigned to the foreground (so your objects of interest).
You might want to have a look on the Principles page for some introduction into this basic segmentation as a starting point.
@ smith_robertj and biovoxxel
thanks for your help