Abstract: A method for illuminating samples in microscopic imaging methods, wherein a number m of different wavelengths ?i, with m>I and i=I, . . . , m, is selected for the illumination. For each of the wavelengths ?i a target phase function ??i(x, y, ?i) is predefined, wherein x and y denote spatial coordinates in a plane perpendicular to an optical axis z and each target phase function ??i(x, y, ?i) is effective only for the corresponding wavelength ?i. The target phase functions ??i are predefined depending on the structure of the sample and/or the beam shape and/or illumination light structure to be impressed on the light used for illumination. A total phase mask is then produced which realises all target phase functions ??i(x, y, ?i). This total phase mask is then illuminated simultaneously or successively with coherent light of wavelengths ?i such that the predefined structure of the illumination light is generated in the region of the sample.

Abstract: An illumination apparatus for illuminating a sample plane and a sample that is optionally arranged therein along an illumination beam path includes: a first light source (for outputting light of at least one first wavelength (?photo), a second light source for outputting light of at least one second wavelength (?exc), and a diffraction grating in the illumination beam path between the first and second light sources and the sample plane. Light of the first wavelength (?photo) is not diffracted by the diffraction grating, and light of the second wavelength (?exc) is diffracted due to the effect of the diffraction grating. The illumination apparatus can be used in a microscope.

Abstract: An optical arrangement for light beam shaping in a light microscope has a first and a second liquid crystal region, each of which has a plurality of independently switchable liquid crystal elements with which a phase of incident light is changeable in a settable manner. A first polarization beam splitter is arranged in such a way that incident light is split in a polarization-dependent manner into reflection light, which is reflected in the direction of the first liquid crystal region, and transmission light, which is transmitted in the direction of the second liquid crystal region. The first or a second polarization beam splitter is arranged such that the reflection light and transmission light are combined onto a common beam path after phase modulation by means of the liquid crystal regions.

Abstract: A single plane illumination microscope having an illumination optical system for illuminating a sample located on a sample carrier in a medium, and which is parallel to a planar reference surface. The sample is illuminated by a light sheet via an illumination light path. A detection optical system has a detection beam path. The optical axes of the illumination and detection optical systems each define an angle that is not equal to zero degrees along with the normal to the reference surface. A barrier layer system includes at least one layer of a given material having a given thickness and separates the medium from the illumination and detection optical systems. A base area of the barrier layer system is in contact with the region that is accessible for illumination and detection activities, said base area running parallel to the reference surface.

Abstract: An optical arrangement for light beam shaping for a light microscope has a first and a second liquid crystal region or lifting micromirror region, each of which has a plurality of independently switchable liquid crystal elements or mirrors with which a phase of incident light is changeable in a settable manner, an input-/output-coupling polarization beam splitter, a polarization beam splitter arranged between the input-/output-coupling polarization beam splitter and the liquid crystal regions or lifting micromirror regions such that the polarization beam splitter separates the light coming from the input-/output-coupling polarization beam splitter in a polarization-dependent manner into a first partial beam, which is directed to the first liquid crystal region or lifting micromirror region, and into a second partial beam, which is directed to the second liquid crystal region or lifting micromirror region, and that the polarization beam splitter combines the two partial beams returning from the liquid crystal

Abstract: A microscopy method, and related microscope, including producing illumination radiation and directing it at a focus. The illumination radiation is switched temporally between at least two modes, such that focus modulation is effected at which temporally varying and mutually different mode fields of the illumination radiation are produced in the focus. The focus is guided at least over regions of a sample to be examined, wherein detection radiation in the sample is or may be brought about by the illumination radiation in the focus at least at a point of origin. The detection radiation is captured in a manner assigned to the at least one point of origin. In addition to the illumination radiation, at least one disexcitation beam of rays of disexcitation radiation is directed at the focus. The disexcitation radiation prevents the detection radiation from being brought about in the region that is illuminated by the disexcitation radiation.

Abstract: An optical assembly that is designed for positioning in a beam path of a light microscope having means for providing structured illuminating light in a sample plane of the light microscope, so that structured illuminating light can be generated in different orientations. The optical assembly has an adjustable deflector in order to deflect an incident light bundle onto one of several beam paths in a selectable manner. Beam splitting devices are located in the beam paths in order to split the light bundle of the respective beam paths into partial light bundles, which are spatially separated from each other. Beam guides are provided for each of the partial light bundles, and guide the partial light bundles to a pupil plane. The beam guides are arranged in such a way that the partial light bundles that belong to the same beam path form a light spot pattern in the pupil plane; and that the light spot patterns of different beam paths in the pupil plane are different from each other.

Abstract: In a high-resolution spectrally selective scanning microscopy of a sample, the sample is excited with illumination radiation in order to emit fluorescence radiation such that the illumination radiation is bundled into an illumination spot in or on the sample. The illumination spot is diffraction-limited in at least one spatial direction and has a minimum extension in said spatial direction. Fluorescence radiation emitted from the illumination spot is imaged into a diffraction image lying on an image plane in a diffraction-limited manner and is detected with a spatial resolution which resolves a structure of a diffraction image of the fluorescence radiation emitted from the illumination spot. The illumination spot is moved into different scanning positions. An individual image is generated for each scanning position, in a diffraction-limited manner onto a detector.

Abstract: In high-resolution scanning microscopy, a sample is excited by illumination radiation to emit fluorescence radiation in such a way that the illumination radiation is focused at a point in or on the sample to form a diffraction-limited illumination spot. 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 structure of the diffraction image. The sample is scanned by means of different scanning positions with an increment of less than half the diameter of the illumination spot. An image of the sample is generated from the data of the surface 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 light microscope comprises: a structuring optical unit comprising a waveguide chip for providing a structured illumination; an input selection device for variably directing light to one of several inputs of the waveguide chip; the waveguide chip further comprising a light guide path following each of the inputs; each light guide path divides into several path divisions; and each path division leads to one output of the wave-guide chip. The outputs of the waveguide chip can be arranged at a pupil plane of the light microscope, and an exit direction of light from the outputs is transverse to a plane defined by the waveguide chip. A method for providing structured illumination light using the light microscope is also described.

Abstract: An arrangement for increasing resolution of a laser scanning microscope has a simplified adjustment and lower susceptibility to errors. The pupil beam from the laser scanning microscope is coupled into a shortened common path interferometer, to make wavefronts of a pupil image mirrored at at least one axis and wavefronts of an unchanged pupil image interfere. The area of a pupil from the pupil beam is split into two complementary portions P and Q producing two partial beams separately supplied to at least one beam deflection means by total-internal reflection along the common path interferometer. The light of the interferometer branches from transmitted light of the one interferometer branch and reflected light of the other interferometer branch is made to interfere at a partly transmissive beam splitter layer to cause constructive interference C and destructive interference D of the wavefronts from the two different portions P and Q of the pupil.

Abstract: An arrangement for TIRF microscopy, having an illumination optical unit with an illumination objective for illuminating a specimen on a specimen carrier in a specimen plane via an illumination beam path. An optical axis of the illumination objective includes an illumination angle that differs from zero with the normal of the specimen plane. A detection optical unit with a detection objective in a detection beam path includes a detection angle that differs from zero between an optical axis thereof and the normal of the specimen plane. A transition element between the specimen carrier and both objectives is arranged both in the illumination beam path and in the detection beam path. The transition element corrects aberrations that arise on account of the passage through media with different refractive indices of radiation to be detected and/or radiation for illuminating the specimen.

Abstract: A method for operating a light microscope with structured illumination includes: providing illumination patterns by means of a structuring device which splits impinging light into at least three coherent beam parts which correspond to a ?1., 0., and +1. diffraction orders of light; generating different phases of the illumination patterns by setting different phase values for the beam parts with phase shifters; and recording at least one microscope image for each of the illumination patterns and calculating a high resolution image from the microscope images. Phase shifters are provided not only for the beam parts of the ?1. and +1. diffraction orders but also at least one phase shifter for the beam part of the 0. diffraction order. At least two different phase values ?0 are set with the at least one phase shifter for the 0. diffraction order to provide a plurality of illumination patterns with different phases.

Abstract: A single plane illumination microscope having an illumination optical system for illuminating a sample located on a sample carrier in a medium, and which is parallel to a planar reference surface. The sample is illuminated by a light sheet via an illumination light path. A detection optical system has a detection beam path. The optical axes of the illumination and detection optical systems each define an angle that is not equal to zero degrees along with the normal to the reference surface. A barrier layer system includes at least one layer of a given material having a given thickness and separates the medium from the illumination and detection optical systems. A base area of the barrier layer system is in contact with the region that is accessible for illumination and detection activities, said base area running parallel to the reference surface.

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 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: 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: 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, 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.