Abstract: The invention relates to the high-resolution spectrally selective scanning microscopy of a sample. The sample is excited with illumination radiation in order to emit fluorescence radiation such that the illumination radiation is bundled into an illumination spot in or on the sample. The illumination spot is diffraction-limited in at least one spatial direction and has a minimum extension in said spatial direction. Fluorescence radiation emitted from the illumination spot is imaged into a diffraction image lying on an image plane in a diffraction-limited manner and is detected with a spatial resolution which resolves a structure of a diffraction image of the fluorescence radiation emitted from the illumination spot. The illumination spot is moved into different scanning positions relative to the sample in increments which are smaller than half the minimum extension of the illumination spot.

Abstract: A method for the optical detection of an illuminated specimen, wherein the illuminating light impinges in a spatially structured manner in at least one plane on the specimen and several images of the specimen are acquired by a detector in different positions of the structure on the specimen. An optical sectional image and/or an image with enhanced resolution is then calculated. The method includes generating a diffraction pattern in the direction of the specimen in or near the pupil of the objective lens or in a plane conjugate to the pupil. A phase plate with regions of varying phase delays is dedicated to the diffraction pattern in or near the pupil of the objective lens or in a plane conjugate to said pupil, and different phase angles of the illuminating light are set.

Abstract: A light scanning microscope with an illumination module switchable between an illumination with m number of spots and an illumination with n number of spots, a deflecting unit which moves the m or n spots in a predetermined sample region, and a detector module for confocal and spectrally resolved detection of the sample radiation. The detector module has a confocal diaphragm unit, a splitting unit which is arranged downstream of the confocal diaphragm unit, a detector, and an imaging unit which images the partial beams on the detector in a spatially separated manner. The confocal diaphragm unit is switchable between a confocal diaphragm with exactly m apertures for m-spot illumination and a confocal diaphragm with n apertures for n-spot illumination. The splitting unit has a first beam path for m-spot illumination and a second beam path for n-spot illumination. The splitting unit is switchable between the two beam paths.

Abstract: An arrangement and method for light sheet microscopy. The arrangement has an illumination apparatus for producing a light sheet for illuminating a stripe of a specimen, and has a detection apparatus for detecting fluorescence radiation emitted by the specimen. The recording speed of the arrangement is increased by an illumination apparatus which is configured to produce at least one further light sheet that is arranged parallel to a first light sheet for illuminating a further stripe of the specimen, and advantageously by a detection apparatus which is configured for the simultaneous detection of the fluorescence radiation excited by the light sheets that are arranged parallel to one another.

Abstract: A method and a configuration for the depth-resolved optical detection of a specimen, in which a specimen or a part of the specimen is scanned by means of preferably linear illumination. The illumination of the specimen is periodically structured in the focus in at least one spatial direction. Light coming from the specimen is detected and images of the specimen are generated. At least one optical sectional image and/or one image with enhanced resolution is calculated through the specimen. Images are repeatedly acquired and sectional images are repeatedly blended while changing the orientation of the linear illumination relative to the specimen and/or spatial intervals between lines exposed to detection light from the illuminated specimen region are generated for the line-by-line non-descanned detection on an area detector or a camera and/or, during a scan, light is further deflected upstream of the detector through the line in the direction of the scan of the specimen.

Abstract: A method for high-resolution 3D fluorescence microscopy, wherein fluorescence emitters in a sample are repeatedly excited to emit fluorescence, and still images are produced of the sample by means of a microscope. The microscope has an imaging beam path with an optical resolution and a focal plane, wherein the fluorescence emitters are excited to emit fluorescence in such a manner that at least a subset of the fluorescence emitters is isolated in each still image so that the images of these fluorescence emitters can be separated in the still images within the optical resolution. The positions of the fluorescence emitters are localized in the generated still images, from the images of the isolated fluorescence emitters, with a location accuracy exceeding the optical resolution, and a high-resolution composite image is generated therefrom.

Abstract: A microscope and method for high resolution scanning microscopy of a sample, having: an illumination device for the purpose of illuminating the sample, an imaging device for the purpose of scanning at least one point or linear spot across the sample and of imaging the point or linear spot into a diffraction-limited, static single image below a reproduction scale in a detection plane. A detector device for detecting the single image in the detection plane for various scan positions is also provided. An evaluation device for the purpose of evaluating a diffraction structure of the single image for the scan positions is provided. The detector device has a detector array which has pixels and which is larger than the single image.

Abstract: In a microscope for high resolution scanning microscopy of a sample, said microscope comprising—an illumination device for illuminating the sample, —an imaging device for scanning at least one point spot or line spot across the sample and for imaging the point spot or line spot into a diffraction-limited, stationary single image with magnification into a detection plane, —a detector device for detecting the single image in the detection plane for different scanning positions with a spatial resolution, which, taking into consideration the magnification, is at least twice as high as a full width at half maximum of the diffraction-limited single image, —an evaluation device for evaluating a diffraction pattern of the single image for the scanning positions from data of the detector device and for generating an image of the sample, said image having a resolution that is increased beyond the diffraction limit, provision is made for—the detector device to have a detector array, which has pixels and is larger than t

Abstract: The invention relates to a device and a method for imaging a sample (2) arranged in an object plane (1). Such a device comprises an optical relay system (3) which images an area of the sample (2) from the object plane (1) into an intermediate image plane (4). Here, the object plane (1) and the intermediate image plane (4) with an optical axis (5) of the relay system (3) include an angle different from 90°. The optical relay system (3) is composed of several lenses. The device also comprises an optical imaging system (6) with an objective, the optical axis (7) of which lies perpendicularly on the intermediate image plane (4) and which is focused on the intermediate image plane (4), with the result that the object plane (1) can be imaged undistorted onto a detector (8).

Abstract: In a microscope for high resolution scanning microscopy of a sample, said microscope comprising—an illumination device for illuminating the sample, —an imaging device for scanning at least one point spot or line spot across the sample and for imaging the point spot or line spot into a diffraction-limited, stationary single image with magnification into a detection plane, —a detector device for detecting the single image in the detection plane for different scanning positions with a spatial resolution, which, taking into consideration the magnification, is at least twice as high as a full width at half maximum of the diffraction-limited single image, —an evaluation device for evaluating a diffraction pattern of the single image for the scanning positions from data of the detector device and for generating an image of the sample, said image having a resolution that is increased beyond the diffraction limit, provision is made for—the detector device to have a detector array, which has pixels and is larger than t

Abstract: An optical device comprises a light source and a detector, and also a sample holder, which is configured to fix an object in the optical path of light. A scanning optical unit is configured, for a multiplicity of scanning positions, in each case selectively to direct light incident from different angular ranges from the object onto the detector. On the basis of a three-dimensional light field represented by corresponding measurement data of the multiplicity of scanning positions, a spatially resolved imaging of the object is generated, said imaging comprising at least two images from different object planes of the object.

Abstract: A microscope including a sample carrier configured to support a sample. Excitation light illuminates the sample via an excitation beam path, Detection light from the sample is guided to detection means via a detection beam path. Through an objective arranged along the optical axis, excitation light is guided in direction of the sample carrier and detection light coming from the sample is guided in direction of the detection means. Beam-splitting means separate excitation light and detection light. Also provided are means for generating a light sheet from excitation light, and means for illuminating the sample with this light sheet. The light sheet lies in a plane at a nonzero angle to the optical axis. The means for illuminating the sample include an optical-deflecting device arranged on or at the sample carrier, which deflects excitation light from the objective into the plane of the light sheet via an optically active surface.

Abstract: A microscope including a sample carrier configured to support a sample. Excitation light illuminates the sample via an excitation beam path. Detection light from the sample is guided to detection means via a detection beam path. Through an objective arranged along the optical axis, excitation light is guided in direction of the sample carrier and detection light coming from the sample is guided in direction of the detection means. Beam-splitting means separate excitation light and detection light. Also provided are means for generating a light sheet from excitation light, and means for illuminating the sample with this light sheet. The light sheet lies in a plane at a nonzero angle to the optical axis. The means for illuminating the sample include an optical-deflecting device arranged on or at the sample carrier, which deflects excitation light from the objective into the plane of the light sheet via an optically active surface.

Abstract: In a microscope for high resolution scanning microscopy of a sample, said microscope comprising—an illumination device for illuminating the sample, —an imaging device for scanning at least one point spot or line spot across the sample and for imaging the point spot or line spot into a diffraction-limited, stationary single image with magnification into a detection plane, —a detector device for detecting the single image in the detection plane for different scanning positions with a spatial resolution, which, taking into consideration the magnification, is at least twice as high as a full width at half maximum of the diffraction-limited single image, —an evaluation device for evaluating a diffraction pattern of the single image for the scanning positions from data of the detector device and for generating an image of the sample, said image having a resolution that is increased beyond the diffraction limit, provision is made for—the detector device to have a detector array, which has pixels and is larger than t

Abstract: A method for high-resolution PAL microscopy, wherein a sample field is imaged on a detector surface of a detector, the sample field is imaged into an image field which is smaller than the detector surface, and the image field on the detector surface is shifted, so that the same sample field is imaged in different positions located adjacent to one another on the image field in order to determine information about changes in the sample field.

Abstract: A microscope which makes possible a spectrally-flexible excitation and detection of fluorescence in an economical manner. For this purpose, means for frequency conversion are arranged in the common beam path and a filter for excitation light is arranged in addition to the main beam splitter in the detection beam path. The frequency conversion achieves a spectral delimitation between illumination light, which is emitted by the light source, and excitation light which brings about fluorescence excitation in the specimen. Because the frequency conversion takes place in the common beam path after the main beam splitter, it is possible for both a spatial separation of illumination light, and excitation light and fluorescent light (detection light) emitted by the specimen, to be carried out in an economical manner at the main beamsplitter according to spectral bands because of the spectral difference between illumination light and excitation light.

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: In structured illumination microscopy, the multiple recording of images with different phase positions of the structuring requires a high stability in the optical arrangement and sample throughout the entire measuring process. Also, the structuring must be projected into the sample in a highly homogeneous manner. The current invention optimizes recording of individual images in order to achieve the best possible resolution in the result image even in problematic samples. An optimization of this kind can be carried out in different ways, for example, by determining an optimal adjustment for at least one illumination parameter or recording parameter or by pulsed illumination such that an excitation from a triplet state of the fluorescent dye to a higher triplet state is reduced, or by illuminating the sample with depletion light for depopulating a triplet state of the fluorescent dye, which reduces bleaching.

Abstract: An apparatus for imaging a sample arranged in a first medium in an object plane. The apparatus includes an optical transmission system which images the sample in the object plane in an intermediate image in an intermediate image plane. The object plane and the intermediate image plane form an angle not equal to 90° with an optical axis of the transmission system. The apparatus further comprises an optical imaging system having an objective. The object plane may be imaged on a detector without distortion. The optical transmission system is symmetrical with respect to a pupil plane, the object plane, and the intermediate image plane to satisfy the Scheimpflug condition. The intermediate image lies in a second medium having a refractive index virtually identical to that of the first medium. A lens group of a subsystem arranged closest to the sample or intermediate image comprises at least one catadioptric assembly.

Abstract: A method for wavelength-resolving and high spatial resolution fluorescence microscopy in which fluorescence labels in a sample are repeatedly excited to emit fluorescence radiation and frames including images of isolated labels are produced with a microscope. The positions of the images of the isolated fluorescing labels are localized with a localization precision exceeding the optical resolution of the imaging beam path of the microscope. The imaging beam path of the microscope has a diffractive element which, during the imaging, diffracts the image of the sample comprising the isolated fluorescing labels into a first diffraction order so that each frame contains the first diffraction order images of the isolated fluorescing labels. A parameter of the first diffraction order images of the isolated fluorescing labels is evaluated and an indication of the wavelength of the isolated fluorescing labels is derived from this evaluated parameter.