Abstract: The invention relates to an illumination apparatus for a microscope, a microscope and a method for operating the illumination apparatus. The illumination apparatus has a sample space for holding a sample that is to be illuminated, and at least one laser light source. An objective for the directional emission of laser radiation of a first wavelength along a first optical axis that is directed into the sample space, and with a cover of the sample space by which the sample space is delimited at least on one of its sides. The cover further has a layer that is either impenetrable for the laser radiation over a blocking angle range of the illumination angle and is transmissive for radiation of a second wavelength over a transmitted light angle range, or has a controllable layer that, in a first control state, is transparent for radiation of the second wavelength and, in a second control state, is impenetrable for the laser radiation of the first wavelength.

Abstract: An arrangement for TIRF microscopy, having an illumination optical unit with an illumination objective for illuminating a specimen on a specimen carrier in a specimen plane via an illumination beam path. An optical axis of the illumination objective includes an illumination angle that differs from zero with the normal of the specimen plane. A detection optical unit with a detection objective in a detection beam path includes a detection angle that differs from zero between an optical axis thereof and the normal of the specimen plane. A transition element between the specimen carrier and both objectives is arranged both in the illumination beam path and in the detection beam path. The transition element corrects aberrations that arise on account of the passage through media with different refractive indices of radiation to be detected and/or radiation for illuminating the specimen.

Abstract: Method for calibrating a phase mask in a beam path of an optical device with the steps: the phase mask is actuated successively with different patterns of grey levels, wherein a first grey level of a first quantity of segments remains constant and a second grey level of a second quantity of segments is varied from one pattern to the next, light of the optical device impinges on the phase mask, at least one part of the total intensity of the light in the beam path is measured downstream of the phase mask for the different patterns, and a characteristic of the measured intensity is obtained in dependence on the second grey level, a relationship between the second grey level and a phase shift, being imprinted by the phase mask, is obtained from the characteristic and an actuation of the phase mask is calibrated based on the obtained relationship.

Abstract: The invention relates to a light microscope for examining microscopic objects with high throughput. The microscope comprises a light source for illuminating a measuring zone, a sample vessel, in which the microscopic objects can be successively moved into the measuring zone, and a detection device for measuring detection light, which originates from a microscopic object located in the measuring zone. According to the invention, the microscope is characterized in that the imaging means comprise a detection lens having a stationary front optics and movable focusing optics, wherein the focusing optics is arranged behind the front optics and in front of an intermediate image plane, and can be adjusted for the height adjustment of a detection plane. The invention further relates to a corresponding microscopy method.

Abstract: A method for providing an overview image of an object that is arranged in a sample plane and includes generating a light sheet, capturing detection light coming from the sample plane, imaging the captured detection light by means of a detector in a detection plane in the form of image data of at least one captured image, wherein the captured image extends in an image plane that is inclined with respect to the sample plane, capturing a number of images of at least one region of the object, in the form of an inclined stack, and transforming the inclined stack to a normalized Z-stack, in which image data of the captured images are assigned with correct orientation with respect to the reference axis. A maximum intensity projection in the normalized Z-stack, wherein a resulting overview image is generated by way of selected image points being imaged as a virtual projection into a projection plane that is parallel to the image plane of the detector.

Abstract: A device and a method for imaging a sample arranged in an object plane. The device includes an optical relay system that images an area of the sample from the object plane into an intermediate image plane. The device may also include an optical imaging system with an objective having an optical axis that lies perpendicularly on the intermediate image plan, and which is focused on the intermediate image plane, with the result that the object plane can be imaged undistorted onto a detector. The device also can include an illumination apparatus for illuminating the sample with a light sheet, wherein the light sheet lies essentially in the object plane and defines an illumination direction, and wherein the normal of the object plane defines a detection direction.

Abstract: A method for adjusting a specimen holder in the beam path of a microscope, in which at least one beam of an illumination radiation is directed onto the specimen holder; a component of the illumination radiation reflected by the specimen holder is captured by means of a detector and measurement values of the captured illumination radiation are ascertained. A current actual manner of positioning of the specimen holder in relation to the beam path is established depending on the measurement values; the established actual manner of positioning is compared to an intended manner of positioning, and control commands for modifying the actual manner of positioning are produced, the execution of which causes the specimen holder to be moved into the intended manner of positioning.

Abstract: The invention relates to a light microscope for examining microscopic objects with high throughput. The microscope comprises a light source for illuminating a measuring zone, a sample vessel, in which the microscopic objects can be successively moved into the measuring zone, and a detection device for measuring detection light, which originates from a microscopic object located in the measuring zone. According to the invention, the microscope is characterized in that the imaging means comprise a detection lens having a stationary front optics and movable focusing optics, wherein the focusing optics is arranged behind the front optics and in front of an intermediate image plane, and can be adjusted for the height adjustment of a detection plane. The invention further relates to a corresponding microscopy method.

Abstract: A method for capturing image data and for determining the thickness of a specimen holder in the beam path of a microscope. The specimen holder is transparent to illumination radiation and embodied to receive a specimen. The specimen holder, which has a first side face and a second side face, is arranged in a specimen plane and the first side face and second side face of the specimen holder are aligned parallel to the specimen plane. At least one beam of the illumination radiation is directed onto the aligned specimen holder along a first optical axis at a first illumination angle and at least two measured values of a reflected component of the illumination radiation or at least two measurement values of a detection radiation caused by the illumination radiation are captured. Depending on the at least two captured measurement values, a spacing of the first and second side face in relation to one another in the direction of the Z-axis is established as a thickness.

Abstract: A single plane illumination microscope having an illumination optical system for illuminating a sample located on a sample carrier in a medium, and which is parallel to a planar reference surface. The sample is illuminated by a light sheet via an illumination light path. A detection optical system has a detection beam path. The optical axes of the illumination and detection optical systems each define an angle that is not equal to zero degrees along with the normal to the reference surface. A barrier layer system includes at least one layer of a given material having a given thickness and separates the medium from the illumination and detection optical systems. A base area of the barrier layer system is in contact with the region that is accessible for illumination and detection activities, said base area running parallel to the reference surface.

Abstract: A method for capturing image data and for determining the thickness of a specimen holder in the beam path of a microscope. The specimen holder is transparent to illumination radiation and embodied to receive a specimen. The specimen holder, which has a first side face and a second side face, is arranged in a specimen plane and the first side face and second side face of the specimen holder are aligned parallel to the specimen plane. At least one beam of the illumination radiation is directed onto the aligned specimen holder along a first optical axis at a first illumination angle and at least two measured values of a reflected component of the illumination radiation or at least two measurement values of a detection radiation caused by the illumination radiation are captured. Depending on the at least two captured measurement values, a spacing of the first and second side face in relation to one another in the direction of the Z-axis is established as a thickness.

Abstract: An arrangement for microscopy, having an illumination optical unit with an illumination objective for illuminating a specimen situated on a specimen carrier in a specimen region of a specimen plane via an illumination beam path. An optical axis of the illumination objective lies in a plane which includes an illumination angle that differs from zero with the normal of a specimen plane, in respect of which the specimen carrier is aligned, and the illumination is implemented in the plane. Further, a detection optical unit with a detection objective is located in a detection beam path. The optical axis of the detection objective includes a detection angle that differs from zero with the normal of the specimen plane. The illumination objective and/or the detection objective comprises an illumination correction element arranged in the beam path and/or a detection correction element.

Abstract: A microscope including an illumination objective with a first optical axis, embodied to produce a light sheet, and a detection objective with a second optical axis, embodied to detect light coming from the specimen plane. The illumination objective and the detection objective are aligned relative to one another and the specimen plane so that the first and second optical axes intersect in the specimen plane and include a substantially right angle therebetween. The optical axes each include an angle which differs from zero with a reference axis directed orthogonal to the specimen plane. An overview illumination apparatus for wide-field illumination of the specimen plane, includes an illumination optical unit with a third optical axis. The characterizing feature is that the detection objective is provided to detect both light from the light sheet and light from the illumination optical unit. A method is also provided for operating a light sheet microscope.

Abstract: An arrangement and method for supplying immersion media and a method for setting optical parameters of a medium, includes a media supply unit for the controlled supply of a medium or of a mixture into a contact region between an optical lens and a specimen slide, on which a specimen may be arranged. An image capture unit is provided for capturing image data on the basis of detection radiation from the object space along a detection beam path extending through the contact region. An evaluation unit is provided to establish current image parameters on the basis of captured image data, to compare said current image parameters to intended image parameters and to establish a desired mixing ratio of at least two components of the medium depending on the comparison.

Abstract: A method for examining a specimen via light sheet microscopy includes selecting several illumination wavelengths for the specimen. To structure the illumination light, a predefined phase distribution is impressed on the phase-selective element and a predefined aperture structure is impressed on an aperture in the aperture plane. The phase-selective element is then illuminated in an intermediate image plane in an illumination beam path with illumination light, which is structured by the phase-selective element. The structured illumination light is imaged into an aperture plane arranged downstream of the phase-selective element. The aperture structure is adapted such that the zero orders of the structured illumination light in the aperture plane are substantially removed. The specimen is illuminated with the structured light sheet in the light sheet plane. Light emitted by the specimen is detected in a detection direction which forms an angle different from zero with the light sheet plane.

Abstract: A method for operating a microscopy arrangement, and a microscopy arrangement, having a first microscope and at least one further microscope, wherein each of the microscopes have a respective optical axis. The respective optical axes do not coincide. The method provides a three-dimensional reference coordinate system being set; a carrier apparatus, that is embodied in the arrangement to receive and hold a specimen carrier is introduced into a specimen plane of the first microscope that is intersected by the optical axis and onto the optical axis of the first microscope; a reference point is set on the optical axis of the first microscope; the carrier apparatus is delivered to the further microscope, wherein the current coordinates of the reference point are continuously captured and compared to the coordinates of the optical axis of the at least one further microscope; and the reference point is brought onto the optical axis of the at least one further microscope.

Abstract: A method for examining a specimen via light sheet microscopy includes selecting several illumination wavelengths for the specimen. To structure the illumination light, a predefined phase distribution is impressed on the phase-selective element and a predefined aperture structure is impressed on an aperture in the aperture plane. The phase-selective element is then illuminated in an intermediate image plane in an illumination beam path with illumination light, which is structured by the phase-selective element. The structured illumination light is imaged into an aperture plane arranged downstream of the phase-selective element. The aperture structure is adapted such that the zero orders of the structured illumination light in the aperture plane are substantially removed. The specimen is illuminated with the structured light sheet in the light sheet plane. Light emitted by the specimen is detected in a detection direction which forms an angle different from zero with the light sheet plane.

Abstract: An optical transmission system configured to image a selected region of a sample arranged in a first medium in an object plane on or in a sample carrier, which includes plane-parallel plate, from the object plane into an intermediate image plane in a second medium. The plane-parallel plate is located between the optical transmission system and the sample during the imaging. The object plane and the intermediate image plane form an angle between 0° and 90° with an optical axis of the transmission system. The optical transmission system is positioned relative to region of the sample such that the sample is located within the focal length of the lens of the optical transmission system closest to the sample. The intermediate image plane and the object plane are located on the same side of the optical transmission system, and the intermediate image is a virtual image.

Abstract: An arrangement for light sheet microscopy that includes a means for scanning a sample volume with a light sheet, which includes an angle ??90° with the optical axis of an objective. The light sheet passes through the entire sample volume in the propagation direction, and the depth of field Sobj of the objective is less than the optical-axis depth T of this sample volume. An optical device, disposed downstream of the objective, increases the depth of field Sobj to a depth of field Seff?the depth T of this sample volume. The arrangement also includes a means for positioning the sample volume within the region of the depth of field Seff. A spatially resolving optoelectronic area sensor is disposed downstream of the optical device, and hardware and software are provided to generate sample-volume images from the electronic image signals output by the area sensor.

Abstract: A three-dimensional high-resolution localization microscopy method including illuminating a sample by excitation radiation to excite fluorescence markers in the sample to luminesce, and imaging the sample in an image frame via imaging optics along an imaging direction, wherein the image frame contains images of the luminescing fluorescence markers, and the imaging optics have a plane of focus and an optical resolution. The excitation step and imaging steps are repeated multiple times to generate a plurality of image frames, wherein the excitation steps are performed to isolate the images of the luminescing fluorescence markers in each image frame for at least some of the luminescing fluorescence markers. The location of the corresponding fluorescence marker is determined in each instance in the generated plurality of image frames from the isolated images of the luminescing fluorescence markers, and a highly resolved total image is generated from the locations determined in this way.