A method for improving image quality is provided. The method includes receiving image data of a substrate, wherein the image data is generated by imaging the substrate, and an image is generated from the image data. The method further includes generating a background representation from a background noise portion of the image, wherein the background portion includes signal information undesired for further processing and generating a background subtracted image by subtracting the background representation from the image. In this way, a separate background image is not needed to subtract the background from the image including the regions-of-interest to improve image quality.