Abstract: A method for determining roughness data and/or topography data of surfaces in material microscopy, particularly from flat samples, based on a shearing polarization interferometrical sequence with a microscopic “TIC” module (“Total Interference Contrast Module”) of a microscope, wherein the method can be carried out both polychromatically and monochromatically. At least two tilted wave fronts are generated, which after reflection or transmission on a sample generate two images of said sample in the form of fringe patterns, said images being offset relative to one another and interfering with one another, from which roughness values and height topographies of the surface of the sample are determined by application of image evaluation.

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 preparing for and carrying out the microscopic recording of image stacks of a sample from various orientation angles, wherein the sample is retained in a sample retainer defining a sample coordinate system. The sample retainer can be translated and rotated in a space spanned by a detection coordinate system. In a first embodiment, for at least two different orientation angles, a projection image is recorded in the detection coordinate system and a first sample volume and a second sample volume are determined in the sample coordinate system. Then the intersection volume of the two sample volumes is determined in the sample coordinate system. Parameters necessary for image stack recording are determined for all other orientation angles based on the intersection volume.

Abstract: Examination of a microscopic specimen is described. Height information for a respective plurality of lateral regions of the specimen is obtained from each of multiple specimen recordings, in which the height information of each specimen recording is limited to a respective height measurement range and the height measurement ranges of different specimen recordings are different. An overall image is calculated from the specimen recordings, in which overall image height information of the different specimen recordings is combined.

Abstract: The invention relates to a device for microscopy, with at least one light source for providing illumination light, with a detection unit for detecting light radiated back from a sample, with a microscopy optical unit for guiding illumination light onto the sample and for guiding light radiated back from the sample in the direction of the detection unit and with, arranged in an illumination beam path, an excitation mask (40) for providing structured illumination. The device is characterized in that the excitation mask is a spatially structured filter, which is transparent to light with a first physical property and which impresses spatial structure onto light with a second physical property that is different from the first physical property. The invention moreover relates to a method for microscopy.

Abstract: A microscope including an imaging objective for imaging a sample on a detector and means for illuminating the sample with a light sheet in the focus plane of the imaging objective. The illumination means includes an illumination source which emits coherent light, and Bessel optics which generate at least two plane waves from the light beam and give propagation directions for the plane waves. The propagation direction of each of the plane waves encloses an acute angle with the focus plane in each instance, the magnitude of the acute angle being identical for each of the plane waves, so that the plane waves undergo constructive interference in the focus plane so that a light sheet is generated. Similarly, the illumination means can also include an optical element by which a rotationally symmetric Bessel beam is generated from the light beam for dynamic generation of a light sheet.

Abstract: The invention relates to a light microscope comprising a polychromatic light source for emitting illumination light in the direction of a sample, focussing means for focussing illumination light onto the sample, wherein the focussing means, for generating a depth resolution, have a longitudinal chromatic aberration, and a detection device, which comprises a two-dimensional array of detector elements, for detecting sample light coming from the sample. According to the invention, the light microscope is characterized in that, for detecting both confocal portions and non-confocal portions of the sample light, a beam path from the sample to the detection device is free of elements for completely masking out non-confocal portions. In addition, the invention relates to a method for image recording using a light microscope.

Abstract: A microscopy device, particularly for use in an imaging fluorescence lifetime microscopy method is provided. The microscopy device comprises an illumination means for generating an illumination beam, an imaging detector for spatially resolved acquisition of an emission radiation emitted by an object to be examined, an illumination beam path between the illumination means and the object to be examined, and a detection beam path between the object to be examined and the detector. The illumination beam path comprises illumination optics which are designed to generate a light sheet of illumination radiation extending transverse to the axis of the illumination beam path, wherein the axis of the detection beam path is oriented substantially perpendicular to a section plane of the light sheet and of the object to be examined. The illumination means comprise a pulsed laser.

Abstract: A family of microscopes include an illumination device which produces a planar light sheet along an illumination axis of an illumination beam path and a transverse axis normal to the illumination axis. A detection device detects light emitted from the sample region along an axis of detection of a detection beam path. The illumination and detection axes as well as the transverse axis and the axis of detection being oriented relative each other at an angle unequal to zero. A light sheet generator also produces rotationally symmetrical light and includes structure and control for rapidly scanning the sample region along the transverse axis. The illumination device includes a second light sheet generator having a first astigmatically active optical element with at least one astigmatic lens for producing a static sheet of light. Selection elements used to select either the first or the second light sheet or both together to produce the sheet of light.

Abstract: A method for determining roughness data and/or topography data of surfaces in material microscopy, particularly from flat samples, based on a shearing polarization interferometrical sequence with a microscopic “TIC” module (“Total Interference Contrast Module”) of a microscope, wherein the method can be carried out both polychromatically and monochromatically. At least two tilted wave fronts are generated, which after reflection or transmission on a sample generate two images of said sample in the form of fringe patterns, said images being offset relative to one another and interfering with one another, from which roughness values and height topographies of the surface of the sample are determined by application of image evaluation.

Abstract: A method for SPIM microscopy, wherein the sample is moved continuously, and a plurality of images are taken at time intervals by means of a detection arrangement during the movement. The image capture duration or exposure time is dimensioned such that the movement path of the sample lies within a predetermined resolution range of the detection objective. The speed of the sample movement is determined and set by the image capture duration or exposure time and/or the distortion of the point spread function generated by the sample movement of the sample. The image blur is corrected computationally by the respective image capture duration and the movement speed. A sharp image is generated in this way. The actual optical section thickness of the light sheet is determined from the light sheet thickness, and the movement speed is determined therefrom and from user settings.

Abstract: A microscope including an illumination device for a light sheet having an approximately planar extension along an illumination axis of an illumination beam path with a transverse axis to the illumination axis. Light emitted from the sample region on axis of detection the illumination axis and the axis of detection as well as the transverse axis and the axis of detection being oriented relative at an non-zero angle. The illumination device includes structure deflecting light to different beam path and thus produces an additional sheet of light, together illuminating the sample region on common illumination axis, and with switches between beam paths. Detection device has a detection lens system for light from by the sample region. The switches include a rapidly switching element with a time <10 ms, a predetermined integration of the surface detector synchronized so the sample region is illuminated twice during integration.

Abstract: The invention is directed to method for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement. According to the invention, the method mentioned above has the following method steps: a sample is introduced into a transparent medium, preferably agarose gel, which is initially liquid; the medium is changed from the liquid state to the solid state, wherein the sample is fixated within the medium, but the transparency of the medium is retained; the solidified medium is positioned in the microscope arrangement in such a way that the sample enclosed therein is situated in the detection area of the objective. Further, a device is proposed for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement.

Abstract: A sample holding system for a microscope, including a sample chamber which has an upper opening and is filled with an immersion liquid, and in which a sample embedded in a transparent embedding compound is placed in a holder. Translatory movement of the sample in relation to a detection objective of the microscope, and rotating the sample about an essentially vertical rotational axis in a plane forming an angle different from zero with the optical axis of the detection objective is included. Rotating the sample includes a rotational drive provided with a magnetic coupling or a belt drive and/or toothed wheel rotational drive arranged above the sample chamber.

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: The invention is directed to an optical arrangement for photomanipulation of a sample comprising a sample holder for receiving the sample, an illumination device comprising an illumination light source and an illumination beam path for illuminating the sample with a light sheet. It further comprises a detection device for detecting light that is radiated from the sample and imaging optics which image the sample on the detection device by means of an imaging objective in an imaging beam path, wherein the light sheet is substantially planar in the focus of the imaging objective, and wherein the imaging objective has an optical axis which intersects the plane of the light sheet at an angle different from zero. Further, the arrangement also has means for photomanipulation of the sample.

Abstract: The invention is directed to a sample holder for a microscope. The sample holder comprises a sample chamber which is filled with an immersion liquid and in which a sample is located. The sample chamber has an upper opening. It further comprises means for translating the sample relative to a detection objective of the microscope, and means for rotating the sample around an axis of rotation extending in a substantially horizontal plane which encloses an angle other than zero degrees with the optical axis of the detection objective. In a sample holder of this kind, the sample is embedded in a transparent embedding medium having at least partially a greater solidity than the immersion liquid. Further, the sample chamber has means for horizontally supporting the embedded sample against the effect of gravity.

Abstract: The invention is directed to an optical arrangement with a light source for emitting a light bundle and with optical elements for transforming this light bundle into the shape of a light sheet, particularly suitable for illuminating individual planes of a three-dimensional specimen in selective plane illumination microscopy (SPIM). According to the invention, means are provided for varying the cross section of the light sheet, for varying the length of the light sheet and/or for influencing the direction in which individual beam components extending within the light sheet are directed to the specimen substance. This makes it possible to adapt the geometry of the light sheet to the illumination requirements for observing one and the same specimen plane with a plurality of different objectives and, if required, to reduce shadows occurring within the observed specimen plane as a result of the illumination.