A microscope (10) is described, having an autofocus system (11) for executing a focusing procedure, having a first image sensor (14a), arranged in a first outcoupled beam path (12a), for acquiring a first image (16a); and a second image sensor (14b), arranged in a second outcoupled beam path (12b), for acquiring a second image (16b). The autofocus system (11) is embodied to ascertain a contrast difference based on contrast values of the first image (16a) acquired by the first image sensor (14a) and of the second image (16b) acquired by the second image sensor (14b), and to set a relative location of the focal plane (20) with respect to the object plane (22) based on the ascertained contrast difference, the first and the second image (16a, 16b) each encompassing image information furnished by the first and the second image sensor (14a, 14b) each embodied as an area sensor.

An observation apparatus, an observation method, and an observation program capable of capturing an image by appropriately adjusting a focus regardless of a size of an effective range in which a distance to a cultivation container can be measured within a field of view are provided. An observation apparatus includes an imaging unit 37 that images an observation target in a field of view smaller than an accommodation part 22 at a series of imaging positions and acquires a series of partial images, a measurement unit 38 that measures a distance from the imaging unit 37 to the accommodation part 22, a storage unit 44 that stores shape information of a container 20 and a series of imaging position information, a calculation unit 45 that calculates effective range information indicating an effective range in which the measurement unit 38 is capable of performing measurement before imaging within a field of view of the imaging unit 37 at the imaging positions, based on the shape information and the imaging position information, and a control unit 40 that controls a focus of the imaging unit 37 using a measurement result measured by the measurement unit 38 in the effective range and a measurement result of the measurement unit 38 in a field of view adjacent to the field of view including the effective range in a case where the effective range is smaller than or equal to a threshold value.

A dynamic focusing system (DFS) for an optical device for observing an object. The dynamic focusing system (DFS) has an objective housing (OBH) including a circumferential housing part (CIH) and a light-transmitting window (LTW); a chamber (ICH) filled with an immersion medium having a refractive index substantially equal to the refractive index of the object to be observed; a movable immersed objective (MIO) configured to move along at least one direction so as to vary the focusing depth and being at least partially immersed in the immersion medium. The immersion medium is confined in the chamber (ICH) by a deformable sealing member (DSM) attached to the movable immersed objective (MIO) and to the objective housing (OBH). The deformable sealing member (DSM) is configured to ensure that the volume of the chamber (ICH) remains substantially constant upon displacement of the movable immersed objective (MIO) relative to the objective housing (OBH).

A method is useful for recording a microscopic fluorescence image of a sample region. An objective directs a laser beam on the sample region having boundary surface(s). A relative distance between the objective and the sample region is altered along an optical axis of the objective to effectuate respective, different relative distances. A respective set of pixel intensity values are effectuated on sensor pixels of an image sensor by the laser beam and transmitted back through the objective is captured for a respective relative distance. A respective focus metric is determined for a respective relative distance based on the respective set of pixel intensity values captured for the respective relative distance. A preferred relative distance is determined based on the determined focus metrics. The preferred relative distance is set, the sample region is illuminated with excitation radiation and the microscopic fluorescence image is captured via the image sensor.

System and method using a projected reference to guide adjustment of a correction optic. In an exemplary method, a reference may be projected onto an imaging detector by propagation of light generally along an optical axis that extends from the reference, through an objective, to a surface of a sample holder, and from the surface, back through the objective, to the imaging detector. The light may propagate through an off-axis aperture located upstream of the imaging detector and spaced from the optical axis. A plurality of images of the reference may be captured using the imaging detector, and with a correction optic at two or more different settings. A setting for the correction optic may be selected based on the plurality of images, and a sample may be imaged while the correction optic has the selected setting.

We describe in this application systems and methods for autofocusing in imaging mass spectrometry. The present application describes improvements over current IMS and IMC apparatus and methods through an autofocus component including a plurality of apertures in the autofocus system, such as a plurality of apertures arranged in 2 dimensions. As a plurality of apertures is used, the autofocus system provides redundancy in the event that measurement of focus on the sample from the illuminating radiation passed through one or more of the apertures fails so as to reduce the number of unsuccessful autofocus attempts.

An optical system including a first light source, a first lens, and a light splitting module, wherein the light splitting module has a first splitter, a second lens, a first camera and a second camera, the first lens is configured for receiving a first light beam from the first light source and collimating the first light beam onto a sample, and for receiving and collimating a light beam from the sample, the second lens is configured for focusing the collimated light beam from the first lens to the first camera and the second camera, the first splitter is configured for splitting the focused light beam from the second lens into a second light beam and a third light beam, the first camera is configured for receiving the second light beam, and the second camera is configured for receiving the third light beam.

A light sheet microscope includes a sample chamber in which a cover slip or slide is arrangeable, which has a surface that defines a partially reflective interface and which has a further surface that defines a further partially reflective interface. The two interfaces are arranged at different distances from an objective. The light sheet microscope further includes an optical system having the objective facing toward the cover slip or slide, an illumination apparatus, which is designed to generate a light sheet, a sensor, and a processor. The two interfaces are formed in that two optical media are applicable in the sample chamber. The light sheet microscope forms a measuring device for acquiring a measured variable. The sensor is designed to acquire the intensities and/or the incidence locations of the two reflection light beams.

Method and system for on-chip processing to obtain an EDOF image combines interferometry and imaging so the two operations do not interfere with one another but, rather, work together to create an in-focus, true color image of a three-dimensional object. This image has no significant artifacts and requires only limited processing. In addition, a coarse depth map is created in the process which may also be helpful in subsequent usage of the acquired image. A CMOS pixel-array sensor includes circuitry to implement processing at the pixel level.

The present disclosure discloses a method and a system for imaging. The method for imaging objects using the system for imaging. The system for imaging comprises a lens. The objects comprise a first object, a second object and a third object located at different positions on a first preset track. The method for imaging comprises: allowing the lens and the first preset track to move relatively in a first predetermined relationship to acquire a clear image of the third object using the system for imaging without focusing, the first predetermined relationship is determined by a focal plane position of the first object and a focal plane position of the second object. The aforementioned method for imaging is high in imaging efficiency and is capable of fast focusing according to the first predetermined relationship even if focus tracking fails so that the blurring of a photographed image due to defocusing is avoided.