1. A system comprising: a calibration device comprising at least one calibration target, wherein each calibration target comprises a sub-region comprising a number of features with dimensions less than the diffraction limit of a microscope used to image the sub-region of the at least one calibration target, wherein the diffraction limit of the microscope is about 200 nm; and a microscope comprising: an objective configured to collect light output from the sub-region of the at least one calibration target and split the light output from each feature into a plurality of color component beams, each color component beam representing a different color component in a color spectrum of the light output from each feature; one or more detectors configured to generate separate color images for certain color component beams, each color image representing a different color image of the at least one calibration target; and a processor configured to combine the separate color images to form a pre-image and generate a calibrated image based on the pre-image, the calibrated image to show a focused and undistorted image of features of the sub-region.