Abstract: The invention relates to a method for high-resolution luminescence microscopy of a specimen marked with marker molecules, and to a luminescence microscope for performing the method, wherein the marker molecules can be excited to emit luminescence radiation. The method for luminescence microscopy comprises the excitation and imaging of marker molecules and the transmission of a trigger time and a position of the specimen. An optical recording device images the marker molecules in a capture area and transmits data from the imaging to an image capture circuit. The recording device transmits a time for the imaging to a signal former as a trigger time; the trigger time is then transmitted to a data recorder.

Abstract: A method for wavelength-resolving and high spatial resolution fluorescence microscopy in which fluorescence labels in a sample are repeatedly excited to emit fluorescence radiation and frames including images of isolated labels are produced with a microscope. The positions of the images of the isolated fluorescing labels are localized with a localization precision exceeding the optical resolution of the imaging beam path of the microscope. The imaging beam path of the microscope has a diffractive element which, during the imaging, diffracts the image of the sample comprising the isolated fluorescing labels into a first diffraction order so that each frame contains the first diffraction order images of the isolated fluorescing labels. A parameter of the first diffraction order images of the isolated fluorescing labels is evaluated and an indication of the wavelength of the isolated fluorescing labels is derived from this evaluated parameter.

Abstract: A microscope for high-resolution imaging of a sample in a depth direction and transversely thereto has an excitation beam path for illuminating a sample,—an imaging beam path with an objective and two detectors,—and a phase element. The phase element is situated in a pupil of the imaging beam path and has a different influence on two halves of the pupil cross section. The imaging beam path is split into two partial imaging beam paths downstream of the pupil as seen in the imaging direction, which partial imaging beam paths each lead to one of the two detectors. The two partial imaging beam paths have imaging lengths that differ by a specific wavelength difference such that the two detectors record images of the sample from two different focal planes, which are spaced apart by a distance in the depth direction.

Abstract: The invention relates to a high-resolution microscope and to a method for determining the two- or three-dimensional positions of objects. The microscope and method includes the following: (a) The vertical (Z) position of imaged particles or molecules being determined from the orientation and shape thereof by means of an anamorphic lens, preferably a cylindrical lens, in the imaging, (b) the detection beam path being split into at least two partial detection beam paths having different optical path lengths, which are detected at an offset on a detector, (c) activation or switchover being performed by means of a multi-photon excitation process, preferably two-photon excitation.

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: An optical arrangement for positioning in a beam path of a light microscope, has an optical carrier, on which a first set of optical assemblies for generating structured illumination light of different orientations is arranged. The optical arrangement includes an adjustable deflection device provided for selectably deflecting a light beam to one of the optical assemblies and for deflecting one light beam coming from said optical assembly into the direction of a sample that is to be examined. The invention further relates to a light microscope having an optical arrangement according to the invention.

Abstract: An optical arrangement for being positioned in a beam path of a light microscope, having at least a first and a second optical assembly for providing structured illumination light from incident light. The optical arrangement provides for light to be guided over different beam paths to the various optical assemblies and in the direction of a sample. Electronic control means are provided and designed to illuminate, in each case, a beam path from the different beam paths to different optical assemblies at a point in time, in that at least a first beam combination mirror is provided for guiding light coming from various optical assemblies to a common beam path in the direction of a sample. The first beam combination mirror has reflective areas on which only light from one of the two optical assemblies is incident and has the light-permeable areas of the beam combination mirror in which only light from the other of the optical assemblies is incident.

Abstract: A microscope for raster-free, confocal imaging of a sample arranged in a sample space has an illumination arrangement comprising a light source group having light sources which can be switched on individually, a detector arrangement, a pinhole arrangement which comprises a pinhole array and which has a plurality of pinhole elements which are adjacent to one another, wherein there is one pinhole element provided for each light source, and optics which irradiate each pinhole element with radiation of an individual light source of the light source group and confocally illuminate an individual spot located in the sample space, wherein one of the individual spots is associated with each pinhole element, and the individual spots are adjacent to one another in the sample space with respect to an incidence direction of the radiation, and the optics image the individual spots through the pinhole arrangement confocally on the detector arrangement.

Abstract: A microscope including a sample carrier configured to support a sample. Excitation light illuminates the sample via an excitation beam path, Detection light from the sample is guided to detection means via a detection beam oath. Through an objective arranged along the optical axis, excitation light is guided in direction of the sample carrier and detection light coming from the sample is guided in direction of the detection means. Beam-splitting means separate excitation light and detection light. Also provided are means for generating a light sheet from excitation tight, and means for illuminating the sample with this light sheet. The light sheet lies in a plane at a nonzero angle to the optical axis. The means for illuminating the sample include an optical-deflecting device arranged on or at the sample carrier, which deflects excitation light from the objective into the plane of the light sheet via an optically active surface.

Abstract: Microscope and method for detecting sample light, having at least one illuminating beam which is partially phase-modulated with a modulation frequency along the cross section thereof and a microscope objective for intensity-modulated focusing of the illuminating beam into a sample. The microscope has a detection beam path that has at least one demodulator. At least one electro-optical modulator (EOM) is used for phase modulation of at least a part, preferably half, of the illuminating beam, or different portions or halves of the illuminating beam are modulated differently, preferably anti-phase, by anti-phase control of piezoelectric elements, or acousto-optical modulators for splitting into a plurality of partial beam paths. Optic elements are provided for partial phase modulation of the excitation beam.

Abstract: A microscope or use of a microscope with at least one illumination beam that is phase-modulated along its cross section partially with a modulation frequency, in which an advantageously local excitation of a transition, advantageously a fluorescence excitation in a sample, is performed by means of a pump beam of a first wavelength and the transition is induced and detected advantageously by means of a second beam, advantageously of a second wavelength, or a local heating is generated that is read out from the detection signals by means of a detection unit, advantageously an infrared camera.

Abstract: A method for wavelength-selective and high spatial resolving fluorescence microscopy. In a specimen fluorescence emitters are repeatedly excited and specimen frames are produced with a microscope. The fluorescence emitters are excited to emit fluorescence radiation such that at least a sub-set is isolated in each frame and the positions of the isolated fluorescence emitters are localized with a localization precision exceeding the optical resolution and a high-resolution complete image is produced.

Abstract: A microscope 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 over the sample and of imaging the point or linear spot into a diffraction-limited, static single image below am imaging scale in a detection plane. A detector device for detecting the single image in the detection plane for various scan positions, with a spatial resolution which, taking into account the imaging 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. The amplitude and/or phase of a wavefront influenced by the sample is detected with spatial resolution by means for wavefront detection, and wherein the influence of the sample on the phase is determined by means of a wavefront sensor.

Abstract: A method for high-resolution 3D fluorescence microscopy, wherein fluorescence emitters in a sample are repeatedly excited to emit fluorescence, and still images are produced of the sample by means of a microscope. The microscope has an imaging beam path with an optical resolution and a focal plane, wherein the fluorescence emitters are excited to emit fluorescence in such a manner that at least a subset of the fluorescence emitters is isolated in each still image so that the images of these fluorescence emitters can be separated in the still images within the optical resolution. The positions of the fluorescence emitters are localized in the generated still images, from the images of the isolated fluorescence emitters, with a location accuracy exceeding the optical resolution, and a high-resolution composite image is generated therefrom.

Abstract: A microscope, preferably a laser scanning microscope, with at least one illuminating beam, which in a partial area along the cross-section thereof, is phase-modulated with a modulation frequency. A microscope objective is provided for focusing the illumination beam onto a sample. The microscope further has a detection beam path and at least one demodulation means, wherein a pulsed illumination beam is present. In the illumination beam path upstream of the microscope objective, a first polarization beam splitter is provided, which generates at least first and second partial beam paths that have differing, preferably adjustable, optical paths. A combination element, such as a second pole splitter, for rejoining the partial beams is provided. In one partial beam path, a phase element is provided, which has at least two areas having differing phase interferences.

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 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: The invention relates to a high resolution microscope for three-dimensionally determining the position of objects, in particular individual fluorophores, and preferably for the high spatial resolution luminescence microscopy of a sample, which is marked with marker molecules that can be activated or switched using a signal such that they can be induced to emit certain luminescent radiation only in the activated state. The object is represented by means of an imaging system, preferably the microscope lens, on a surface detector consisting of individual detector elements.

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: In a sample, fluorescence emitters are repeatedly excited to emit fluorescence, and still images are produced of the sample by means of a microscope. At least a subset of the fluorescence emitters is isolated in each still image. The positions of the fluorescence emitters are localized in the still images with a location accuracy exceeding the optical resolution. A high-resolution composite image is generated therefrom. An adaptive mirror is arranged in the imaging beam path, and is adjusted in such a manner that it produces an astigmatism when at least one of the still images is produced. As a result, still images with astigmatism are captured. Depth position information for the fluorescence emitters is derived from the rotational asymmetry. The adaptive mirror is additionally adjusted in such a manner that it does not produce any astigmatism when some of the still images are produced.