I made images with the videomicroscope and ZEN images.
I would like to measure a bone volume (area), but I have a problem, the value is not good.
I have area differences between the images before and after threshold (not on the top of the picture) but but when I measure by drawing a square. I don’t think the threshold is supposed to change the area of the image.
I specify that I tried by putting the real dimensions (set scale: 1pxl = 0.1615µm) or by putting in pixel. I still have problems with measured areas that are wrong. I tried directly with the original image (.czi) or in tiff/jpeg without results.
I also have another problem. When I measure areas using the wand tracing tool or polygon selections. I get very different things.
For example on this image, I drew the shapes 1 (left) and 3 (right) with the wand tracing tool and the shapes 2 (left) and 4 (right) with polygon selections.
The values of 1 and 2 or 3 and 4 are not at all comparable. Moreover, the area of the right shape is more important than the left one, but with the wand tool, I get the opposite.
Can you help me ?