System for creating microscopic digital montage images

An imaging apparatus, comprising:

• a motorized stage;
• a camera having a lens directed toward said motorized stage;
• and
• a processor coupled to said camera, wherein said processor contains instructions which, when executed by said processor, cause said processor to:
• select at least three points of a sample adjacent said motorized stage;
• determine a stage position for each selected point;
• focus said camera on each selected point;
• determine an object distance from the camera lens to the sample at each selected point;
• develop a focus surface based on stage position and object distance for the at least three selected points;
• determine for a selected point whether an error function based on the difference between the object distance at the selected point and the object distance of the surface at that point is greater than a predetermined limit;
• select an additional point from an area in which the point has an error function greater than a predetermined limit;
• determine stage position for the additional selected point;
• focus said camera on the additional selected point;
• determine object distance from the camera lens to the sample at the additional selected point;
• and
• develop a focus surface based on stage position and object distance for the selected points.

A system for creating a high throughput montage image of a microscope slide, the system comprising:

• an optical system that comprises at least one camera, a motorized stage for moving a slide while an image of the slide is captured, a pulsed light illumination system that optically stops motion on the motorized stage while allowing continuous physical movement of the motorized stage, and a stage position detector that controls firing of the pulsed light illumination system at predetermined positions of the motorized stage;
• a first component that identifies sample regions on the slide in the optical system and determines locations of the sample on the slide, wherein the first component uses information about the locations to generate control parameters for the motorized stage and the camera;
• a second component that uses the control parameters to ensure that a high-quality montage image is captured;
• and
• means for capturing a montage image, using the second component, by maintaining motion of the motorized stage and synchronization of the optical system, thereby enabling accurate focus control of optical elements without requiring the stage to be stopped and refocused at each tile location.

A method for creating a high throughput montage image of microscope slides, the method comprising:

• placing a slide to be imaged in a slide holder on a motorized stage;
• capturing a low resolution image of the slide;
• identifying sample locations within the image;
• generating control parameters to scan regions of interest under microscopic optics;
• capturing a high resolution montage image by enabling accurate focus control of optical elements without requiring that the motorized stage be stopped and refocused at each tile location in the montage image;
• controlling a tiling process by moving the motorized stage;
• capturing image tiles with precise alignment by executing a strobe illumination system whenever a stage position sensor determines that the motorized stage has moved to a neighboring field of view of a camera;
• scanning each row of locations identified to contain the sample for the sample;
• and
• removing the slide and inserting another slide to be imaged.