I’m new to this forum so many apologies if a similar post has been made and since resolved.
I have a series of images which I’ve cropped so the tiff file assigns a black background that surrounds my images. I want to measure the mean gray value in the tissue but I think imagej recognises the black background as a gray object (measures it as 1 when converted to 8 bit grayscale). As I have a series of images, my plan is write a macro so I can then batch process the images to obtain my measurements. I’ve uploaded my image for reference, which is a section tissue stained with luxol fast blue and cresyl violet. I use colour deconvolution to separate out the stains as I want to quantify the LFB intensity. This automatically converts the image to 8 bit, then I convert to grayscale using lookup tables.
So far, I’ve resorted to using tswayne’s solution by subtracting that 1 pixel from my images as a makeshift solution:
Any further advice would be much appreciated!