Abstract: A method and apparatus for light field microscopy, wherein the following method steps are performed: a) measuring an image data record of a sample using a light field arrangement; b) creating at least one partial data record from the image data record; c) reconstructing a three-dimensional object from the partial data record created in step b).

Abstract: A method of image evaluation when performing SIM microscopy on a sample includes: A) providing n raw images of the sample, which were each generated by illuminating the sample with an individually positioned SIM illumination pattern and imaging the sample in accordance with a point spread function, B) providing (S1) n illumination pattern functions, which each describe one of the individually positioned SIM illumination patterns, C) providing (S1) the point spread function and D) Carrying out an iteration method, which includes following iteration steps a) to e), as follows: a) providing an estimated image of the sample, b) generating simulated raw images, in each case by image processing of the estimated image using the point spread function and one of the n illumination pattern functions such that n simulated raw images are obtained, c) assigning each of the n simulated raw images to that of the n provided raw images which was generated by the illumination pattern that corresponds to the illumination patter

Abstract: A method for performing SIM microscopy on a sample, includes: generating n raw images of the sample, in each case by illuminating the sample using the same SIM illumination pattern albeit with an individual positioning for each raw image, wherein p orders of diffraction are assigned to the SIM illumination pattern, and generating an image of the sample from the n raw images. An image reconstruction is carried out using the orders of diffraction, wherein t highest orders of diffraction are suppressed during the image reconstruction and n=p?t applies.

Abstract: A description is given of a method for increasing resolution in microscopy, comprising providing at least one recorded sample image (22) which was generated by means of a microscope (2), providing a point spread function which characterizes an imaging behaviour of the microscope (2), and calculating a sample image with increased resolution from the recorded sample image (22), wherein the calculating is effected in an iteration process (S4) which repeatedly passes through an iteration loop (S4a; S4b) and which determines a correction image (24.0-24.

Abstract: The invention relates to a localization microscopy method for localizing signal sources. Here, at least once for each pixel of a detector, values of an error parameter are ascertained and stored in a calibration data record in a manner assigned to the relevant pixel. Captured image data are used to identify regions of origin of signal sources and fit a point spread function to the pixel values of the respective regions of origin. The respective signal source is localized on the basis of the point spread function. The pixel-specific error parameter of each pixel can be compared to a threshold. If the threshold is exceeded, these pixels are either ignored or replaced by means of interpolation when fitting the point spread function. In addition or as an alternative thereto, the real noise performance of the pixels is ascertained and corrected on the basis of derived pixel-specific error parameters.

Abstract: In a method for deconvolving image data, image data of an object are captured with a number n of confocal beam paths. The image data are converted into resultant image data by means of a point spread function. The resultant image data are deconvolved again in the frequency domain using a deconvolution function, wherein the deconvolution function contains the formation of at least a number n of sum terms and a Wiener parameter w. The results of the sum terms are stored in retrievable fashion; the Wiener parameter W is modified at least once proceeding from its original value and the deconvolution is carried out by means of the deconvolution function with the modified Wiener parameter w and the stored results of the sum terms.

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 method of image evaluation when performing SIM microscopy on a sample includes: A) providing n raw images of the sample, which were each generated by illuminating the sample with an individually positioned SIM illumination pattern and imaging the sample in accordance with a point spread function, B) providing (S1) n illumination pattern functions, which each describe one of the individually positioned SIM illumination patterns, C) providing (S1) the point spread function and D) Carrying out an iteration method, which includes following iteration steps a) to e), as follows: a) providing an estimated image of the sample, b) generating simulated raw images, in each case by image processing of the estimated image using the point spread function and one of the n illumination pattern functions such that n simulated raw images are obtained, c) assigning each of the n simulated raw images to that of the n provided raw images which was generated by the illumination pattern that corresponds to the illumination patter

Abstract: Method for super-resolution evaluation of microscope images illuminated in a structured manner and microscope having structured illumination. The resolution can be improved laterally by a factor of up to two using conventional linear structured Illumination (SIM). If a non-linear iterative method is used for the purpose of deconvolution, the achievable resolution can be improved beyond the theoretical limit. However, the known methods only achieve a small amount of increase. The novel method is intended to make improved resolution or improved contrast possible. If a PSF/OTF that is manipulated (individually for each order) in the same (or in a corresponding) way as the relevant order spatial frequency spectrum is used during the re-weighting in the spatial frequency domain (for the deconvolution), the actually achievable resolution can be nearly doubled in comparison with the conventional SIM, both in one-stage and in two-stage variants.

Abstract: A method for performing SIM microscopy on a sample, includes: generating n raw images of the sample, in each case by illuminating the sample using the same SIM illumination pattern albeit with an individual positioning for each raw image, wherein p orders of diffraction are assigned to the SIM illumination pattern, and generating an image of the sample from the n raw images. An image reconstruction is carried out using the orders of diffraction, wherein t highest orders of diffraction are suppressed during the image reconstruction and n=p?t applies.

Abstract: The invention relates to a microscope having an excitation beam path for guiding excitation light, having a laser light source for providing a laser light beam as excitation light and having a scanning apparatus for aligning and moving a focused laser light beam in the entrance pupil of an illumination objective; wherein the laser focus is directed into an entrance point that is offset with respect to the optical axis of the illumination objective; and also having a detection beam path for guiding detection light, comprising a microlens array having a focal plane for generating partial imaged presentations and a detector arranged in the focal plane of the microlens array for capturing the partial imaged presentations. In addition, an evaluation unit for evaluating the captured image signals of the detector in accordance with light-field technology is present. The invention additionally relates to a method for microscopic image generation.

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: A method for high-resolution scanning microscopy of a sample in which a sample is illuminated at a point in or on the sample by means of illumination radiation. The point is imaged along an optical axis and according to a point spread function into a diffraction image on a spatially resolving surface detector that comprises detector pixels in which a diffraction structure of the diffraction image is resolved. The point is displaced relative to the sample in at least two scanning directions and pixel signals are read from the detector pixels in various scanning posi- tions, wherein the pixel signals are respectively assigned to that scanning position at which they were read out and adjacent scanning positions overlap one another and are disposed according to a scanning increment. An image of the sample having a resolution that is increased beyond a resolution limit of the imaging is generated from the read pixel signals and the assigned scanning positions, wherein a deconvolution is carried out.

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 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: A fluctuation-based fluorescence microscopy method, comprising influencing a point-spread function of the imaging of a sample emitting fluorescence radiation using an optical device in dependence on a parameter such that a point emitter is imaged into a representation with two image lobes. The relative positions of the lobes depend on the position of the point emitter relative to the focal plane. Synthetic pixels, smaller than detector pixels, are generated; for each synthetic pixel, pairs of pixel groups are defined among pixels of the detector based on the influencing of the point spread function. Each pair is assigned to an individual value of the parameter. In each frame and for each synthetic pixel, a signal correlation is ascertained and allocated as image brightness to the synthetic pixel for the parameter specification. Subframes for each frame are produced from the synthetic pixels, and a high-resolution sample image is produced from the subframes.

Abstract: The invention relates to a localization microscopy method for localizing signal sources. Here, at least once for each pixel of a detector, values of an error parameter are ascertained and stored in a calibration data record in a manner assigned to the relevant pixel. Captured image data are used to identify regions of origin of signal sources and fit a point spread function to the pixel values of the respective regions of origin. The respective signal source is localized on the basis of the point spread function. The pixel-specific error parameter of each pixel can be compared to a threshold. If the threshold is exceeded, these pixels are either ignored or replaced by means of interpolation when fitting the point spread function. In addition or as an alternative thereto, the real noise performance of the pixels is ascertained and corrected on the basis of derived pixel-specific error parameters.

Abstract: In a method for deconvolving image data, image data of an object are captured with a number n of confocal beam paths. The image data are converted into resultant image data by means of a point spread function. The resultant image data are deconvolved again in the frequency domain using a deconvolution function, wherein the deconvolution function contains the formation of at least a number n of sum terms and a Wiener parameter w. The results of the sum terms are stored in retrievable fashion; the Wiener parameter W is modified at least once proceeding from its original value and the deconvolution is carried out by means of the deconvolution function with the modified Wiener parameter w and the stored results of the sum terms.

Abstract: The invention relates to a method for high-resolution scanning microscopy of a sample M which a) the sample is illuminated at a point in or on the sample by means of illumination radiation, b) the point is imaged along an optical axis and according to a point spread function into a diffraction image on a spatially resolving surface detector that comprises detector pixels in which a diffraction structure of the diffraction image is resolved, c) the point is displaced relative to the sample in at least two scanning directions and pixel signals are read from the detector pixels in various scanning positions, wherein the pixel signals are respectively assigned to that scanning position at which they were read out and adjacent scanning positions overlap one another and are dis posed according to a scanning increment, d) an image of the sample having a resolution that is increased beyond a resolution limit of the imaging is generated from the read pixel signals and the assigned scanning positions, wherein a deconvo

Abstract: A fluctuation-based fluorescence microscopy method, comprising influencing a point-spread function of the imaging of a sample emitting fluorescence radiation using an optical device in dependence on a parameter such that a point emitter is imaged into a representation with two image lobes. The relative positions of the lobes depend on the position of the point emitter relative to the focal plane. Synthetic pixels, smaller than detector pixels, are generated; for each synthetic pixel, pairs of pixel groups are defined among pixels of the detector based on the influencing of the point spread function. Each pair is assigned to an individual value of the parameter. In each frame and for each synthetic pixel, a signal correlation is ascertained and allocated as image brightness to the synthetic pixel for the parameter specification. Subframes for each frame are produced from the synthetic pixels, and a high-resolution sample image is produced from the subframes.