We have developed a macro that performs a range of functions on this image. The raw image has been thresholded and binary has been performed once. Normally it is possible to perform binary again to make the image black and white, in order to skeletonize the image (then perform binary connectivity and anaylse histogram).
With recently redownloading ImageJ the program will not perform binary. Performing convert to mask (next image) appears to do this ok, see image and produces the same result. This is slightly concerning and i was wondering why it was necessary to make this change as if we hadn’t fallen upon the solution by chance I am not sure it would be possible to have worked it out?