A fluorescence microscope, includes an optical system configured to collect fluorescent light emitted from different fluorophore species within a field of view and to focus the fluorescent light for detection, a spectral splitting arrangement configured to split the fluorescent light into at least two spectrally different fluorescent light components, a multi-channel detector system including at least two image sensors configured to detect at least two spatial light intensity distributions based on the at least two spectrally different fluorescent light components, each spatial light intensity distribution representing an image of the object over the field of view, and a processor configured to determine spatial distributions of the different fluorophore species based on a spectral unmixing analysis of each spatial light intensity distribution, wherein the processor is further configured to obtain compensation information and to determine a spatial distribution of each fluorophore species by taking into account the compensation information.

A fluorescence microscope, includes an optical system configured to collect fluorescent light emitted from different fluorophore species within a field of view and to focus the fluorescent light for detection, a spectral splitting arrangement configured to split the fluorescent light into at least two spectrally different fluorescent light components, a multi-channel detector system including at least two image sensors configured to detect at least two spatial light intensity distributions based on the at least two spectrally different fluorescent light components, each spatial light intensity distribution representing an image of the object over the field of view, and a processor configured to determine spatial distributions of the different fluorophore species based on a spectral unmixing analysis of each spatial light intensity distribution, wherein the processor is further configured to obtain compensation information and to determine a spatial distribution of each fluorophore species by taking into account the compensation information.

Resolution of images used in processes such as sequencing by synthesis may be increased by structuring sites that would emit signals in the images to have different elevations. Differences in focus caused by these differences in elevation may be used to filter out background illumination, thereby providing an image in which in focus sites may be resolved even though the separation between any site and its nearest neighbor may be below the diffraction limit of the light that would be emitted.

A microscope objective for imaging a specimen using a microscope, the microscope objective being designed as an air objective for microscopy without an immersion medium or as an oil immersion objective for microscopy with an oil-based immersion medium or as a water immersion objective for microscopy with a water-based immersion medium. The front lens of the microscope objective is provided with a coating which repels an immersion medium and is lipophobic and hydrophobic if the objective is an air objective, only lipophobic if the objective is a water immersion objective, and only hydrophobic if the objective is an oil immersion objective.

A system can be used for imaging volumetric samples using a camera array to capture images under different illumination patterns at different axial planes of focus, and at optionally varying lateral fields of view. The sequence of images taken under the different illumination patterns and optional varying lateral fields of view can be processed to generate an image representation of the sample at a focus plane of the sample. Multiple image representations at different focus planes are assembled to form a 3D volumetric representation of the sample.

An interactive apparatus, system and method for image capturing and projecting is integrated into an optical microscope. An image capturing and projecting unit is operatively connected to a processing unit, the processing unit configured to: (a) receive user generated data; and (b) overlay the user generated data onto an optically viewed image visible through the eyepiece.

An interactive apparatus, system and method for image capturing and projecting is integrated into an optical microscope. An image capturing and projecting unit is operatively connected to a processing unit, the processing unit configured to: (a) receive user generated data; and (b) overlay the user generated data onto an optically viewed image visible through the eyepiece.

A method is useable for determining a refractive index of an optical medium in a microscope, which has an objective facing toward a sample chamber. The optical medium is one of two optical media, which border two opposing surfaces of a cover slip or object carrier in the sample chamber and form two partially reflective interfaces, which are arranged at different distances from the objective. The method includes: deflecting a measurement light beam by the objective with oblique incidence on the cover slip or object carrier; generating two reflection light beams spatially separated from one another by the measurement light beam being partially reflected at each of the interfaces; receiving the two reflection light beams by the objective and conducting them onto a position-sensitive detector; registering intensities by the position-sensitive detector; and determining the refractive index of the optical medium based on the registered intensities.

A method is useable for determining a refractive index of an optical medium in a microscope, which has an objective facing toward a sample chamber. The optical medium is one of two optical media, which border two opposing surfaces of a cover slip or object carrier in the sample chamber and form two partially reflective interfaces, which are arranged at different distances from the objective. The method includes: deflecting a measurement light beam by the objective with oblique incidence on the cover slip or object carrier; generating two reflection light beams spatially separated from one another by the measurement light beam being partially reflected at each of the interfaces; receiving the two reflection light beams by the objective and conducting them onto a position-sensitive detector; registering intensities by the position-sensitive detector; and determining the refractive index of the optical medium based on the registered intensities.

A light sheet microscope includes an object slide, and optical illumination and detection systems. The optical illumination system includes an illumination objective configured to illuminate a first object plane that is oblique relative to a slide plane with a light sheet. The optical detection system includes a detection objective and an image sensor device. The image sensor device is configured to define a first image plane which is orthogonal to an optical axis of the detection objective and to define a second image plane which is tilted relative to the first image plane. The detection objective is configured to image a focal plane onto the first image plane and to image a second object plane of the object onto the second image plane, the focal plane being coincident with the first object plane, and the second object plane being parallel to or coincident with the slide plane.