Abstract: A method can be used for correcting background signals in captured measurement values of analog detectors, wherein measurement values of an object captured over a reference time period are analyzed and characteristic values of captured background signals are determined. What is characteristic of this is that a threshold value is determined on the basis of at least one characteristic value and by applying a calculation specification; the threshold value is applied to captured measurement values of an analog detector, and only those measurement values which are greater than the threshold value are used for a subsequent signal evaluation. A microscope for carrying out the method according to the invention is also provided.

Abstract: The invention relates to an FCS method in which a sample that is to be measured and has fluorescent markers is illuminated with excitation radiation over a bleaching time in order to bleach selected fluorescent markers; and after bleaching has been carried out over at least one measurement period, FCS measurement data of the sample are acquired by illuminating the sample with excitation radiation and by detecting detection radiation brought about by the excitation radiation. The invention is characterized in that during the bleaching time, intensity values of fluorescence radiation that has been brought about by the excitation radiation which is directed at the sample for bleaching purposes are continuously or repeatedly acquired and compared with a threshold value, and the acquisition of the FCS measurement data is started when the threshold value has been reached.

Abstract: A microscopy method involves directing a focused beam of excitation radiation at an examination location of an object to be examined, to create an excitation volume in the object. An overview image is used to identify at least one structure of the object and define at least one of the identified structures as a reference structure. A spatial relationship is defined between the positions of the examination location and the reference structure. Detection radiation coming from the excitation volume is acquired as measurement values over an overall measurement duration, the overall measurement duration being subdivided into a plurality of measurement intervals. At least every second measurement interval is preceded by a comparison of the current position of the focused beam with a current position of the examination location. The positioning of the focused beam is corrected in the case of an inadmissible deviation of the current positions.

Abstract: Apparatus and method for capturing an image having a detection beam path for guiding detection radiation from a sample to a detector having a plurality of detector elements. The detector has no more than ten and, preferably, four or five detector elements; and an evaluation unit, which is configured to carry out an evaluation in accordance with the Airyscan method on the image data captured by means of the detector and which generates a high-resolution image.

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: 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 reducing image artifacts in images of a sample captured by scanning, wherein intensity values of at least two detection regions, denoted as pixels (Pxn), are captured along respectively one row (j) in a first scanning direction. A reconstructed image is produced on the basis of the captured intensity values. According to the invention, the intensity values of the reconstructed image are summed along the rows (j) respectively scanned by a certain pixel (Pxn) and a row sum is formed in each case. A correction value of the pixel (Pxn) is ascertained on the basis of the row sums formed thus and the correction value is applied to the intensity values, captured by means of the pixel (Pxn), of the reconstructed image, as a result of which a corrected image is obtained.

Abstract: A method for the high-resolution scanning microscopy of a specimen where the specimen is illuminated with illuminating radiation such that the illuminating radiation is focused to a diffraction-limited illuminating spot at a point in or on the specimen. The point is projected in a diffraction-limited manner in a diffraction image onto a flat panel detector having pixels. The flat panel detector, owing to the pixels thereof, have a spatial resolution which resolves a diffraction structure of the diffraction image. The point is shifted relative to the specimen into different scanning positions by an increment which is smaller than the diameter of the illuminating spot and a 3D image is generated. The pixels of the flat panel detector are divided into groups. A pre-calculated raw image is calculated for each group and are unfolded three-dimensionally to generate the image of the specimen.

Abstract: A method for high-resolution scanning microscopy of a sample, wherein the sample is illuminated with illumination light such that the illumination light is focused at a point in or on the sample into an illumination spot. The point is imaged into a diffraction image onto an area detector having detector elements. The area detector has a spatial resolution that resolves a diffraction structure of the diffraction image. The sample is here scanned line-wise in a grid made of rows and columns by displacing the point relative to the sample into different scanning positions with an increment width that is smaller than the diameter of the illumination spot. The area detector is read, and an image of the sample is generated from the data of the area detector and from the scanning positions assigned to said data, said image having a resolution that is increased beyond a resolution limit for imaging.

Abstract: A method for the high-resolution scanning microscopy of a specimen where the specimen is illuminated with illuminating radiation such that the illuminating radiation is focused to a diffraction-limited illuminating spot at a point in or on the specimen. The point is projected in a diffraction-limited manner in a diffraction image onto a flat panel detector having pixels. The flat panel detector, owing to the pixels thereof, have a spatial resolution which resolves a diffraction structure of the diffraction image. The point is shifted relative to the specimen into different scanning positions by an increment which is smaller than the diameter of the illuminating spot. The flat panel detector is read, and, from the data of the flat panel detector and from the scanning positions assigned to these data, a 3D image of the specimen is generated.

Abstract: The invention relates to an image capturing apparatus and method having a detection beam path for guiding detection radiation from a sample to a detector having a plurality of detector elements. The detector has no more than ten and, in particular, four or five detector elements; and an evaluation unit is present, which is configured to carry out an evaluation in accordance with the Airyscan method on the image data captured by means of the detector and which generates a high-resolution image.

Abstract: A method for high-resolution scanning microscopy of a sample, wherein the sample is illuminated with illumination light such that the illumination light is focused at a point in or on the sample into an illumination spot. The point is imaged into a diffraction image onto an area detector having detector elements. The area detector has a spatial resolution that resolves a diffraction structure of the diffraction image. The sample is here scanned line-wise in a grid made of rows and columns by displacing the point relative to the sample into different scanning positions with an increment width that is smaller than the diameter of the illumination spot. The area detector is read, and an image of the sample is generated from the data of the area detector and from the scanning positions assigned to said data, said image having a resolution that is increased beyond a resolution limit for imaging.

Abstract: The invention relates to a method for reducing image artifacts in images of a sample captured by scanning, wherein intensity values of at least two detection regions, denoted as pixels (Pxn), are captured along respectively one row (j) in a first scanning direction. A reconstructed image is produced on the basis of the captured intensity values. According to the invention, the intensity values of the reconstructed image are summed along the rows (j) respectively scanned by a certain pixel (Pxn) and a row sum is formed in each case. A correction value of the pixel (Pxn) is ascertained on the basis of the row sums formed thus and the correction value is applied to the intensity values, captured by means of the pixel (Pxn), of the reconstructed image, as a result of which a corrected image is obtained.

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 method for correcting colors of a color reproduction of a digital microscope and a digital microscope are described. In a first step of a method according to the invention, a color image of a sample that is to be examined under the microscope is recorded. When the recording is performed, wavelength-dependent properties of a microscope illumination unit that illuminates the sample are determined in order to describe a state of the microscope illumination unit, in that settings selected at the microscope illumination unit are captured. A set of correction values is determined, which is associated with a state of the microscope illumination unit that is selected in accordance with the state of the microscope illumination unit determined when the recording is performed. In a further step, the colors of the recorded color image of the sample are corrected by applying the correction values of the previously determined set.

Abstract: The present invention relates firstly to a method for producing a microscopic image with an extended depth of field by means of a microscope. In a step of the method, a plurality of microscopic frames of a specimen are acquired from different focus positions, for example with different dimensions of a distance between the specimen and an objective lens of the microscope. In another step, the plurality of microscopic frames are processed so as to form a microscopic image with an extended depth of field. According to the invention, the focus position is continuously changed during the acquisition of at least some of the microscopic frames at a variable speed or with variable acceleration. The invention further relates to a microscope with an objective lens for optically imaging a specimen.

Abstract: A method for correcting colors of a color reproduction of a digital microscope and a digital microscope are described. In a first step of a method according to the invention, a color image of a sample that is to be examined under the microscope is recorded. When the recording is performed, wavelength-dependent properties of a microscope illumination unit that illuminates the sample are determined in order to describe a state of the microscope illumination unit, in that settings selected at the microscope illumination unit are captured. A set of correction values is determined, which is associated with a state of the microscope illumination unit that is selected in accordance with the state of the microscope illumination unit determined when the recording is performed. In a further step, the colors of the recorded color image of the sample are corrected by applying the correction values of the previously determined set.