I am using image J recently to quantify the fluorescent intensity of a contrast on human tissue. I am having difficulty when choosing the threshold of the images. Is there a way that I CAN find an optimal threshold to be used for all images? How can I know that my threshold is not over saturating or under saturating my images? how can I choose a right threshold to light only the contrast not other structures.
In addition, is there a way that I can quantify the intensity of each gridlines in my image? I figured how to add grids but I can’t find a way to measure the intensity in each grid.
I would really appreciate if someone can help me with these issues.