An auto-focusing method for determining an in-focus position of a plurality of wells in at least a portion of a multi-well plate, the method including using a first objective lens having a first magnification to identify, in each of at least three wells of a selected subset of the plurality of wells, an in-focus position of each well with respect to the first objective lens, on the basis of at least three the in-focus positions, computing a plane along which the at least three wells will be in focus with respect to at least one objective lens having a second magnification that is not greater than the first magnification, and using the at least one objective lens to scan, along the plane, at least some of the plurality of wells in the portion of the plate.

Real-time autofocus. In an embodiment, a scanning apparatus includes an imaging sensor, a focusing sensor, an objective lens, and processor(s) configured to analyze image data captured by the imaging and focusing sensors, and move the objective lens. Real-time autofocus during scanning of a sample is achieved by determining a true-Z value for the objective lens for a point on a sample and for each of a plurality of regions on the sample. The true-Z values and/or surfaces calculated therefrom are used to determine a predicted-Z value for an unscanned region of the sample. The objective lens is adjusted to the predicted-Z value at the beginning of the unscanned region. After scanning the region, a true-Z value is determined for the region and compared to the predicted-Z value. A rescan of the region is initiated if the comparison exceeds a predetermined threshold.

A microscope system comprises: a light source; an objective lens; a stage; a two-dimensional image sensor that captures an image of a specimen placed on the stage; a focusing device that changes distance between the objective lens and the stage; and a control circuit, wherein the control circuit executes, during a movement period in which the stage moves in a direction orthogonal to an optical axis of the objective lens, focus control for controlling the focusing device based on focus evaluation information detected during the movement period, and exposure control for controlling an exposure period of the two-dimensional image sensor, and executes light emission control that causes the light source to emit light with different light emission intensities during the exposure period and during a focus evaluation period in which the focus evaluation information is detected.

The invention relates to a beam manipulation device for a scanning microscope, comprising a main colour splitter for coupling excitation light into an illumination beam path and for separating excitation light from detection light of a detection beam path, said device comprising a scanner, preferably positioned on a pupil plane, for scanning the excitation light. The device is characterised in that: an additional optical section is provided comprising optical elements which influence a beam path; at least one pupil plane and/or at least one intermediate image plane is formed in the additional optical section by the optical elements which influence the beam path; and an adjustable selection device is provided for activating either a first beam segment of the illumination and/or detection beam path, or the additional optical section, wherein the first beam segment of the illumination and/or detection beam path does not contain a pupil plane of the illumination and/or detection beam path.

A microscope (10) for detecting images of an object (14) located in an object plane (12) is described, comprising a microscope stand (18); a microscope objective (20); a light source (22) integrated into the microscope stand (18); and a beam splitter (24), integrated into the microscope objective (20), for coupling in a coaxial incident illumination.

Systems and methods for capturing a digital image of a slide using an imaging line sensor and a focusing line sensor. In an embodiment, a beam-splitter is optically coupled to an objective lens and configured to receive one or more images of a portion of a sample through the objective lens. The beam-splitter simultaneously provides a first portion of the one or more images to the focusing sensor and a second portion of the one or more images to the imaging sensor. A processor controls the stage and/or objective lens such that each portion of the one or more images is received by the focusing sensor prior to it being received by the imaging sensor. In this manner, a focus of the objective lens can be controlled using data received from the focusing sensor prior to capturing an image of a portion of the sample using the imaging sensor.

A method is used for optical scanning of at least one object region placed on a transparent specimen holder. The method is as follows: for each sample lateral position of plural predefined sample lateral positions performing a focus determination by: performing laser reflection and using a first camera taking plural first images to determine a reference distance between the specimen holder and an objective lens; performing transmission flash illumination and using a second camera taking plural second images to define a focus distance taking into account the reference distance; after completing the focus determination, determining a focus distance topology across the object region based on the focus distances determined for ail sample lateral positions; and laterally moving the specimen holder and acquiring third images while focusing according to the focus distance topology.

Implementing swept, confocally aligned planar excitation (SCAPE) imaging with asymmetric magnification in the detection arm provides a number of significant advantages. In some preferred embodiments, the asymmetric magnification is achieved using cylindrical lenses in the detection arm that are oriented to increase the magnification of the intermediate image in the width direction but not in the depth direction. SCAPE imaging may also be improved by using an SLM to modify a characteristic of the sheet of excitation light that is projected into the sample. Additional embodiments include a customized version of SCAPE that is optimized for imaging the retina at the back of an eyeball in living subjects.

There are provided a microscope apparatus, an observation method, and a microscope apparatus-control program that can more efficiently perform auto-focus control and can shorten an imaging time in a case where a culture vessel is to be scanned by an image forming optical system and the auto-focus control is to be performed at each observation position. Focus information of a culture vessel is detected by a first displacement sensor and a second displacement sensor while a stage is moved to a scanning measurement position from an initial set position, and an auto-focus control unit performs auto-focus control at every observation position on the basis of the focus information in a case where the stage has been moved to the scanning measurement position.

A method and corresponding optical device to correct spherical aberration in an optical path caused by an electrically tunable lens (ETL) within the optical path. The method includes placing within the optical path and in working relationship with the ETL an aspherical correction lens dimensioned and configured to reduce spherical aberration in a light beam exiting the ETL.