Abstract: Accelerated methods and apparatuses for three-dimensional microscopy with structured illumination, in which focal planes of the sample are focused and each focal plane is illuminated in a plurality of phases sequentially with structured illumination light and the sample light emitted by the sample is recorded in a respective individual image. A resulting image having a resolution that is increased with respect to the individual images is reconstructed from the individual images to produce a super-resolved image stack. By reconstructing a resulting image from individual images of two different focal planes by approximation methods, said resulting image represents a sample plane that is situated between said focal planes, an image stack can be produced in a shorter period with less stress on the sample.

Abstract: In a 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. An individual image is generated for each scanning position, in a diffraction-limited manner onto a detector.

Abstract: For the purposes of high-resolution scanning microscopy, a sample is excited by illumination radiation to emit fluorescent radiation in such a way that the illumination radiation is focused at a point in or on the sample, so as to form a diffraction-limited illumination spot. The point is imaged in a diffraction image on a detector in a diffraction-limited manner, wherein the detector has detector elements and a plurality of location channels which resolve a diffraction structure of the diffraction image. The sample is scanned with various scanning positions with an increment smaller than half the diameter of the illumination spot. An image of the sample with a resolution that is increased beyond a resolution limit of the image is generated from the data of the detector and from the scanning positions associated with these data.

Abstract: A microscope and method for high resolution scanning microscopy of a sample, having an illumination device, 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 is used for detecting the single image in the detection plane for various scan positions, with a location accuracy which, taking into account the reproduction scale in at least one dimension/measurement, is at least twice as high as a full width at half maximum of the diffraction-limited single image.

Abstract: A microscope and method for high resolution scanning microscopy of a sample, having an illumination device, 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 is used for detecting the single image in the detection plane for various scan positions, with a location accuracy which, taking into account the reproduction scale in at least one dimension/measurement, is at least twice as high as a full width at half maximum of the diffraction-limited single image.

Abstract: Microscope and method for high resolution scanning microscopy of a sample, wherein the sample is illuminated; at least one point spot or line spot, which is guided in a scanning manner over the sample, is imaged into a still image; wherein the spot is imaged in a diffraction limited manner into the still image with magnification, and the still image lies still in a plane of detection; the still image is detected for different scan 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 still image, so that a diffraction pattern of the still image is detected; the diffraction pattern of the still image is evaluated for each scan position, and an image of the sample is generated that has a resolution that is increased beyond the diffraction limit, wherein a detector array is provided that has pixels and is larger than the still image; and radiation of the still image from the plane of detecti

Abstract: A method for evaluating signals of fluorescence scanning microscopy with simultaneous excitation and detection of fluorescence in different focal planes of a sample by means of confocal laser scanning microscopy.

Abstract: A method and microscope for high-resolution 2D scanning microscopy of a sample, wherein the sample is illuminated with illumination radiation in such a way that the illumination radiation is focused in or on the sample to form a diffraction-limited illumination spot at a point. The point is imaged in a diffraction-limited manner into a diffraction image on a spatially resolving surface detector, wherein the surface detector has a spatial resolution that resolves a diffraction structure of the diffraction image. Neither an imaging point spread function nor an illumination point spread function is manipulated for producing an asymmetry. The point is displaced relative to the sample into different scanning positions. A 2D image of the sample is produced from the data of the surface detector and from the scanning positions assigned to said data. The 2D image has a resolution that is increased beyond a resolution limit for imaging.

Abstract: For the purposes of high-resolution scanning microscopy, a sample is excited by illumination radiation to emit fluorescent radiation in such a way that the illumination radiation is focused at a point in or on the sample, so as to form a diffraction-limited illumination spot. The point is imaged in a diffraction image on a detector in a diffraction-limited manner, wherein the detector has detector elements and a plurality of location channels which resolve a diffraction structure of the diffraction image. The sample is scanned with various scanning positions with an increment smaller than half the diameter of the illumination spot. An image of the sample with a resolution that is increased beyond a resolution limit of the image is generated from the data of the detector and from the scanning positions associated with these data.

Abstract: A method for high-resolution 3D-localization microscopy of a sample having fluorescence emitters, in which the fluorescence emitters are excited to emit fluorescent radiation and the sample is displayed with spatial resolution in wide-field microscopy. Excitation is caused such that at least some fluorescence emitters are isolated. A three-dimensional localization is determined in a localization analysis, which includes a z-coordinate, x-coordinate as well as a y-coordinate orthogonal thereto, for each isolated fluorescence emitter. A table of localization imprecision is provided. Localization imprecision being determined for each localized fluorescence emitter by accessing the table of localization imprecision.

Abstract: A method for high-resolution 3D-localization microscopy of a sample having fluorescence emitters, in which the fluorescence emitters are excited to emit fluorescent radiation and the sample is displayed with spatial resolution in wide-field microscopy. Excitation is caused such that at least some fluorescence emitters are isolated. A three-dimensional localization is determined in a localization analysis, which includes a z-coordinate, x-coordinate as well as a y-coordinate orthogonal thereto, for each isolated fluorescence emitter. A table of localization imprecision is provided. Localization imprecision being determined for each localized fluorescence emitter by accessing the table of localization imprecision.

Abstract: A method for operating a scanning microscope and for determining point spread functions, with which sample images are recorded with the scanning microscope. The method includes scanning a sample with at least one illuminating light beam; recording at least one sample image with a detector device during a scan by the illuminating light beam; and comprising the point spread function, with which a sample image is recorded, from the at least one sample image. A detector device having receiving elements is used, where the distance between the receiving elements is smaller than a diffraction disk that generates a sample point on the detector device. Detector signals, generated by means of the receiving elements, are read out for each of the different positions of the illuminating light beam on the sample, as a result of which the scanning of the sample allows the detector signals, which are read out, to generate a plurality of sample images.

Abstract: A method for high-resolution 3D-localization microscopy of a sample having fluorescence emitters, in which the fluorescence emitters are excited to emit fluorescent radiation and the sample is displayed with spatial resolution in wide-field microscopy. Excitation is caused such that at least some fluorescence emitters are isolated. A three-dimensional localization is determined in a localization analysis, which includes a z-coordinate, x-coordinate as well as a y-coordinate orthogonal thereto, for each isolated fluorescence emitter. A table of localization imprecision is provided. Localization imprecision being determined for each localized fluorescence emitter by accessing the table of localization imprecision.

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 evaluating signals of fluorescence scanning microscopy with simultaneous excitation and detection of fluorescence in different focal planes of a sample by means of confocal laser scanning microscopy.

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: A microscopy high-resolution scanning method, including exciting a sample with illumination radiation focused at a point to form a diffraction-limited illumination spot so as to emit fluorescence radiation. The point is imaged in a diffraction image on a spatially resolving two-dimensional detector. The sample is scanned at scanning positions with increments that are smaller than half the diameter of the spot. An image of the sample with a resolution increased beyond a resolution limit of the image is generated from the data of the two-dimensional detector and the scanning positions. To discriminate between at least two predetermined wavelength ranges in the fluorescence radiation of the sample, Airy disks corresponding to the wavelength ranges are generated on the two-dimensional detector, the Airy disks being offset laterally from one another such that the diffraction image consists of the mutually offset Airy disks. The Airy disks are evaluated when generating the sample image.

Abstract: A microscopy method for producing a high-resolution image of a sample which includes furnishing the sample with a marker that emits statistically flashing luminescence radiation after excitation, or using a sample that has molecules that emit statistically flashing luminescence radiation after excitation. The sample is excited to luminescence in such a manner that the marker/molecules emit luminescence radiation flashing at a flash rate, wherein—the illumination is structured in such a manner that the flash rate varies locally and—the sample is repeatedly illuminated in different illumination states of the structured illumination. The luminescing sample is repeatedly imaged on a detector in each of the different illumination states.

Abstract: A microscopy method for generating a high-resolution image (55) of a sample (2) comprising the following steps: a) the sample (2) is provided with a marker, which upon excitation emits statistically flashing luminescent radiation, or a sample (2) is used which upon excitation emits locally distributed, statistically flashing luminescent radiation; b) the sample (2) is excited to luminescence by means of structured illumination, wherein the sample is repeatedly illuminated in at least nine different illumination conditions (0.01-0.09) of the structured illumination by realizing at least three rotary positions, and at least three displacement positions per rotary position of the structured illumination; c) the luminescing sample (2) is repeatedly displayed in each of the different illumination conditions on a flat panel detector having pixels, such that an image sequence (44.01-44.09) is obtained for each of the different illumination conditions (0.01-0.

Abstract: A light microscope having a specimen plane, in which a specimen to be examined is positioned, having a light source to emit illuminating light, having optical imaging means to convey the illuminating light into the specimen plane, having a first scanning means, with which an optical path of the illuminating light and the specimen can be moved relative to each other to produce an illumination scanning movement of the illuminating light relative to the specimen, having a detector means to detect specimen light coming from the specimen and having electronic means to produce an image of the specimen based on the specimen light detected by the detector means at different specimen regions. A second scanning means is present, with which it can be adjusted which specimen region can be imaged on a determined detector element.