Abstract: A microscope for microscopy having a detection beam path and an illumination beam path. The microscope includes a filter wheel device, which is arranged in the detection beam path and/or illumination beam path and has a filter wheel, wherein the filter wheel is mounted to rotate about an axis, and wherein the filter wheel is divided into segments. Filters forming a first part-circle are arranged in a first part of the segments, such that said filters are consecutively introduced into the detection beam path or illumination beam path when the filter wheel is rotated. A camera records images at a predefined frequency. The filter wheel device includes a motor-driven shaft that rotates the filter wheel at a predefined rotation frequency. The microscope also comprises a control system for synchronising the image-recording frequency and rotation frequency based on the filters arranged on the filter wheel.

Abstract: An optical microscope includes a first mask that has transmission regions that are separated from one another for the simultaneous generation of a plurality of illumination light beams from illumination light, for example, a first scanning device for generating a scanning motion of the illumination light beams and a sample holder. The optical microscope also includes a second mask with transmission regions separated from one another, which transmission regions are smaller than the transmission regions of the first mask in order to clip the illumination light beams, such that, through the scanning motion of the first scanning device, each of the illumination light beams can be successively passed onto different transmission regions of the second mask, and a second scanning device is provided for generating a scanning motion between the clipped illumination light beams and the sample holder. A method for examining a microscopic sample is also provided.

Abstract: A microscope and a method of microscopy that uses structured illumination, involving imaging a grid structure or periodic light distribution on a sample, wherein by displacing the image of the grid structure, imaging is carried out under different phase positions, and a high-resolution sample image is calculated from the recorded images, characterized in that the grid structure or light distribution is generated by using at least two phase grids arranged one in front of the other, and different orientations of the light distribution perpendicular to the illumination direction are generated by displacing the phase grids relative to one another, with displacement, imaging and calculation being carried out for different orientations.

Abstract: A microscopy method for producing a high-resolution image of a 2-dimensional sample. The method includes exciting statistically blinking fluorophores in a sample by irradiating the sample with illumination radiation, repeatedly imaging the sample onto a spatially resolving detector in an image field that covers only a part of the sample to thereby obtain a frame sequence, generating an image from the frame sequence, the image having a spatial resolution increased beyond the optical resolution limit using a cumulant function, moving the position of the image field on the sample at least once and repeating the imaging and generating steps to obtain one image for each position of the image field, and combining the resultantly obtained images to form a complete image of the sample.

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 blinking luminescent radiation, or a sample (2) is used which upon excitation emits locally distributed, statistically blinking 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 (.01-.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 (.01-.

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.

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 method for generating an image of a sample by a microscopy method including varying local resolution, wherein at least two of the following microscopy methods are combined: laser scanning microscopy, a microscopy method wherein the sample is excited to luminescence by structured line or wide area illumination, and a first microscopy image is generated from the images thus obtained, having increased local resolution greater than the optical resolution of the image, a further microscopy method according to the PAL principle, by which a second microscopy image is generated, indicating geometric locations of marker molecules emitting luminescent radiation at an increased local resolution relative to the optical resolution, and a further microscopy method, wherein the sample is marked using marking molecules suitable for the STED, ESA, or RESOLFT technique, and a third microscopy image is generated of STED, ESA, or RESOLFT, wherein the obtained images are superimposed.

Abstract: Method for enhancing the resolution of a microscope during the detection of an illuminated specimen and a microscope for carrying out the method, wherein in a first position, an illumination pattern is generated on the specimen, the resolution of which is preferably within the range of the attainable optical resolution of the microscope or higher, wherein a relative movement, preferably perpendicular to the direction of illumination, from a first into at least one second position of the illumination pattern on the specimen is generated at least once between the detection and the illumination pattern with a step width smaller than the resolution limit of the microscope and detection and storage of the detection signals take place both in the first and in the second position.

Abstract: The invention relates to a method for operating a microscope in which excitation light is focused on, or beamed to, different points of a specimen, in which an intensity of the excitation light is point-specifically varied and in which an intensity of the light reflected by said specimen in at least one spectral range is measured point-specifically and quantitatively. The method according to the invention is characterized in that the intensity and/or a spectral composition of the excitation light beamed to a specific point of said specimen is automatically adjusted by a regulating device on the basis of information previously gained from measured data of said specimen concerning an estimated or actual intensity of the light reflected in the spectral range by said point such that an integral of the intensity of the light reflected in the spectral range by this point during a pixel dwell time is within a predefined value interval. The invention also relates to a microscope.

Abstract: Method for spatially high-resolution luminescence microscopy in which label molecules in a sample are activated to emit luminescence radiation comprising activating only a subset of the label molecules in the sample, wherein activated label molecules have a distance to the closest activated molecules that is greater or equal to a length which results from a predetermined optical resolution, detecting the luminescence radiation, generating a frame from the luminescence radiation, identifying the geometric locations of the label molecules with a spatial resolution increased above the predetermined optical resolution, repeating the steps and forming a combined image, and controlling the acquisition of the several frames by evaluating at least one of the frames or a group of the frames and modifying at least one variable for subsequent repetitions of the steps of generating frames for combining into an image.

Abstract: A method for high-resolution luminescence microscopy of a sample marked with marking molecules that can be activated to excite particular luminescent radiation, including: repeated activation of a subset of the marking molecules to emit luminescent radiation; repeated imaging of the sample along a depth direction and with a predetermined optical resolution; and producing images from the repeated imaging. Locations of the marking molecules are determined with a spatial resolution that is increased above the predetermined optical resolution. Activation of the marking molecules can be through radiation introduced into multiple regions, each extending along a plane substantially perpendicular to the depth direction. The regions can be arranged so that the regions are behind one another and overlap only partially. Separate images of the sample may be recorded for activation in each of the regions in order to obtain depth information relating to the marking molecules from the separate images.

Abstract: A microscopy method for producing a high-resolution image of a 2-dimensional sample. The method includes exciting statistically blinking fluorophores in a sample by irradiating the sample with illumination radiation, repeatedly imaging the sample onto a spatially resolving detector in an image field that covers only a part of the sample to thereby obtain a frame sequence, generating an image from the frame sequence, the image having a spatial resolution increased beyond the optical resolution limit using a cumulant function, moving the position of the image field on the sample at least once and repeating the imaging and generating steps to obtain one image for each position of the image field, and combining the resultantly obtained images to form a complete image of the sample.

Abstract: The invention relates to a method and a microscope for generating a microscopic image, wherein a) the sample is illuminated in each case by the microscope lens using the TIRF method; and b) the sample is illuminated in a structured fashion in different displacement positions of the structure. The sample light of the method according to a) and b) is detected in each case for generating an image of at least once sample region, wherein the sample images generated according to a) and b) are set off against one another, preferably multiplied, and the result is stored for generating a new sample image.

Abstract: The invention allows a quantitative evaluation of images acquired by microscope having fewer errors and is applicable in connection with high-resolution methods, particular at a high speed. A microscope image is analyzed in which the intensity distributions of fluorescence events have in each instance a diffraction-dependent extent which corresponds to an extent of a point spread function of the microscope and are arranged so as to be spatially non-overlapping, or at least predominantly spatially non-overlapping, in that at least one counter is initialized for every region to be analyzed in the microscope image, at least one fluorescence event is identified in a region to be analyzed in the microscope image, and the counter corresponding to the relevant region is incremented for each fluorescence event identified in the region. The counting results in a dramatic improvement in the signal-to-noise ratio at a high evaluation speed.

Abstract: A microscope system comprises a microscope for data acquisition and a computing device configured to control the microscope during data acquisition and/or to perform data processing of raw data captured by the microscope. The computing device is coupled to an optical output device. The microscope and the computing device are configured to perform the data acquisition and/or data processing based on values that are respectively set for each one of a plurality of adjustable parameters. The computing device selectively outputs graphics data via the optical output device as a function of an adjustable parameter selected from the plurality of adjustable parameters. The output graphics data are assigned to the selected adjustable parameter and represent an affect of the selected adjustable parameter on at least one step of a procedure upon which the data acquisition and/or the data processing is based.

Abstract: A method for generating an image of a sample by a microscopy method including varying local resolution, wherein at least two of the following microscopy methods are combined: laser scanning microscopy, a microscopy method wherein the sample is excited to luminescence by structured line or wide area illumination, and a first microscopy image is generated from the images thus obtained, having increased local resolution greater than the optical resolution of the image, a further microscopy method according to the PAL principle, by which a second microscopy image is generated, indicating geometric locations of marker molecules emitting luminescent radiation at an increased local resolution relative to the optical resolution, and a further microscopy method, wherein the sample is marked using marking molecules suitable for the STED, ESA, or RESOLFT technique, and a third microscopy image is generated of STED, ESA, or RESOLFT, wherein the obtained images are superimposed.

Abstract: Method for enhancing the resolution of a microscope during the detection of an illuminated specimen and a microscope for carrying out the method, wherein in a first position, an illumination pattern is generated on the specimen, the resolution of which is preferably within the range of the attainable optical resolution of the microscope or higher, wherein a relative movement, preferably perpendicular to the direction of illumination, from a first into at least one second position of the illumination pattern on the specimen is generated at least once between the detection and the illumination pattern with a step width smaller than the resolution limit of the microscope and detection and storage of the detection signals take place both in the first and in the second position.

Abstract: Method for spatially high-resolution luminescence microscopy in which label molecules in a sample are activated to emit luminescence radiation comprising activating only a subset of the label molecules in the sample, wherein activated label molecules have a distance to the closest activated molecules that is greater or equal to a length which results from a predetermined optical resolution, detecting the luminescence radiation, generating a frame from the luminescence radiation, identifying the geometric locations of the label molecules with a spatial resolution increased above the predetermined optical resolution, repeating the steps and forming a combined image, and controlling the acquisition of the several frames by evaluating at least one of the frames or a group of the frames and modifying at least one variable for subsequent repetitions of the steps of generating frames for combining into an 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.