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: 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 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 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 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 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: With the different methods of fluorescence correlation spectroscopy, physical and biological transport processes in or between cells in the microscopic range, for example diffusion processes, can be analyzed. For this purpose, correlations of the fluorescence measurement data are determined for different sample regions and mathematical transport models are adapted thereto. Erroneous fluorescence correlation analyses were previously identified on the basis of the properties of the adapted model function parameters and were discarded. The a-priori knowledge necessary for the identification had to be obtained in time-consuming series of tests. With the invention, sample properties can be determined in a simpler, quicker and more exact way from fluorescence correlations.

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 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 method for high-resolution 3D-localization microscopy of a sample having fluorescence emitters, in which the fluorescence emitters in the sample are excited to emit fluorescent radiation and the sample is displayed with spatial resolution in wide-field microscopy. Excitation is caused such that in reference to the spatial resolution at least some fluorescence emitters are isolated. A three-dimensional localization is determined in a localization analysis, which includes in the depth direction of the display a z-coordinate and a x-coordinate as well as a y-coordinate orthogonal in reference thereto, for each isolated fluorescence emitter showing a precision exceeding the local resolution. A table of localization imprecision is provided, which states the imprecision of the localization, regarding its z-coordinate as a function of the z-coordinate and a number of photons collected during imaging in the wide-field microscopy.

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: 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 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: 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: 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: With the different methods of fluorescence correlation spectroscopy, physical and biological transport processes in or between cells in the microscopic range, for example diffusion processes, can be analyzed. For this purpose, correlations of the fluorescence measurement data are determined for different sample regions and mathematical transport models are adapted thereto. Erroneous fluorescence correlation analyses were previously identified on the basis of the properties of the adapted model function parameters and were discarded. The a-priori knowledge necessary for the identification had to be obtained in time-consuming series of tests. With the invention, sample properties can be determined in a simpler, quicker and more exact way from fluorescence correlations.