Differential contrast comparison between monochrome and pseudo-H&E fresh tissue imaging to aid diagnosis of gigascale monochromatic tumor margin surface images

Recent developments in fluorescence microscopy have allowed the acquisition of pathology-like images from fresh tissue in close to real time. Monochromatic imaging can improve imaging speed by twofold. However, a few limitations need to be addressed, such as structure contrast and image interpretation. Comparing pseudo-H&E to monochrome images obtained from the same tissue preparations can help to train humans/machines to interpret monochromatic images. For this purpose, we acquired images using a custom-built structured illumination microscope of consecutive microscopic slides, fixed tissue and bovine muscle, and 110 fresh prostate surfaces labeled with acridine orange (63) or SYBR Gold and Eosin (47).