Self-Calibrating and Directional Focusing Systems and Methods for Infinity Corrected Microscopes

A system for autofocusing an objective lens in a microscope system comprising:

• an image capturing device;
• an objective lens;
• a decentered aperture disposed in an optical path between the objective lens and an image plane of the image capturing device;
• and
• a controller that performs an autofocus calibration process that includes:
• causing the objective lens to move along an axis of travel;
• operating the image capturing device to capture a plurality of reference images, wherein each reference image is captured when the objective lens is positioned at a corresponding z-position of a plurality of z-positions along the axis of travel and the optical path is at least partially occluded by the decentered aperture;
• analyzing the plurality of reference images to develop a plurality of pattern locations, wherein each pattern location represents a position of a pattern formed on the image plane when a corresponding reference image was captured, and wherein at least one reference image of the plurality of reference images is associated with a best focus position;
• and
• positioning the objective lens in accordance with the best focus position and the plurality of pattern locations.