Abstract: An arrangement for microscopy, having an illumination optical unit with an illumination objective for illuminating a specimen situated on a specimen carrier in a specimen region of a specimen plane via an illumination beam path. An optical axis of the illumination objective lies in a plane which includes an illumination angle that differs from zero with the normal of a specimen plane, in respect of which the specimen carrier is aligned, and the illumination is implemented in the plane. Further, a detection optical unit with a detection objective is located in a detection beam path. The optical axis of the detection objective includes a detection angle that differs from zero with the normal of the specimen plane. The illumination objective and/or the detection objective comprises an illumination correction element arranged in the beam path and/or a detection correction element.

Abstract: A microscope including an illumination objective with a first optical axis, embodied to produce a light sheet, and a detection objective with a second optical axis, embodied to detect light coming from the specimen plane. The illumination objective and the detection objective are aligned relative to one another and the specimen plane so that the first and second optical axes intersect in the specimen plane and include a substantially right angle therebetween. The optical axes each include an angle which differs from zero with a reference axis directed orthogonal to the specimen plane. An overview illumination apparatus for wide-field illumination of the specimen plane, includes an illumination optical unit with a third optical axis. The characterizing feature is that the detection objective is provided to detect both light from the light sheet and light from the illumination optical unit. A method is also provided for operating a light sheet microscope.

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 method for examining a specimen via light sheet microscopy includes selecting several illumination wavelengths for the specimen. To structure the illumination light, a predefined phase distribution is impressed on the phase-selective element and a predefined aperture structure is impressed on an aperture in the aperture plane. The phase-selective element is then illuminated in an intermediate image plane in an illumination beam path with illumination light, which is structured by the phase-selective element. The structured illumination light is imaged into an aperture plane arranged downstream of the phase-selective element. The aperture structure is adapted such that the zero orders of the structured illumination light in the aperture plane are substantially removed. The specimen is illuminated with the structured light sheet in the light sheet plane. Light emitted by the specimen is detected in a detection direction which forms an angle different from zero with the light sheet plane.

Abstract: A method for operating a microscopy arrangement, and a microscopy arrangement, having a first microscope and at least one further microscope, wherein each of the microscopes have a respective optical axis. The respective optical axes do not coincide. The method provides a three-dimensional reference coordinate system being set; a carrier apparatus, that is embodied in the arrangement to receive and hold a specimen carrier is introduced into a specimen plane of the first microscope that is intersected by the optical axis and onto the optical axis of the first microscope; a reference point is set on the optical axis of the first microscope; the carrier apparatus is delivered to the further microscope, wherein the current coordinates of the reference point are continuously captured and compared to the coordinates of the optical axis of the at least one further microscope; and the reference point is brought onto the optical axis of the at least one further microscope.

Abstract: With the method of the invention a wavelength-dependent refractive index of a specimen medium, which is examined with a light microscope, is determined. With the light microscope, a specimen measurement at the specimen medium which has an unknown refractive index is performed, wherein illumination light is radiated to the specimen medium and detection light coming from the specimen medium is measured. With the specimen measurement, a specimen measurement focus position of the illumination and/or detection light is measured. Using a mathematical model, in which a focus position of illumination and/or detection light is defined in dependence of a refractive index of a medium, the refractive index of the specimen medium is derived from the specimen measurement focus position. Furthermore, a light microscope for carrying out the method is described.

Abstract: A method for examining a specimen via light sheet microscopy includes selecting several illumination wavelengths for the specimen. To structure the illumination light, a predefined phase distribution is impressed on the phase-selective element and a predefined aperture structure is impressed on an aperture in the aperture plane. The phase-selective element is then illuminated in an intermediate image plane in an illumination beam path with illumination light, which is structured by the phase-selective element. The structured illumination light is imaged into an aperture plane arranged downstream of the phase-selective element. The aperture structure is adapted such that the zero orders of the structured illumination light in the aperture plane are substantially removed. The specimen is illuminated with the structured light sheet in the light sheet plane. Light emitted by the specimen is detected in a detection direction which forms an angle different from zero with the light sheet plane.

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: An optical transmission system configured to image a selected region of a sample arranged in a first medium in an object plane on or in a sample carrier, which includes plane-parallel plate, from the object plane into an intermediate image plane in a second medium. The plane-parallel plate is located between the optical transmission system and the sample during the imaging. The object plane and the intermediate image plane form an angle between 0° and 90° with an optical axis of the transmission system. The optical transmission system is positioned relative to region of the sample such that the sample is located within the focal length of the lens of the optical transmission system closest to the sample. The intermediate image plane and the object plane are located on the same side of the optical transmission system, and the intermediate image is a virtual image.

Abstract: An arrangement for light sheet microscopy that includes a means for scanning a sample volume with a light sheet, which includes an angle ??90° with the optical axis of an objective. The light sheet passes through the entire sample volume in the propagation direction, and the depth of field Sobj of the objective is less than the optical-axis depth T of this sample volume. An optical device, disposed downstream of the objective, increases the depth of field Sobj to a depth of field Seff?the depth T of this sample volume. The arrangement also includes a means for positioning the sample volume within the region of the depth of field Seff. A spatially resolving optoelectronic area sensor is disposed downstream of the optical device, and hardware and software are provided to generate sample-volume images from the electronic image signals output by the area 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: An arrangement and method for light sheet microscopy. The arrangement has an illumination apparatus for producing a light sheet for illuminating a stripe of a specimen, and has a detection apparatus for detecting fluorescence radiation emitted by the specimen. The recording speed of the arrangement is increased by an illumination apparatus which is configured to produce at least one further light sheet that is arranged parallel to a first light sheet for illuminating a further stripe of the specimen, and advantageously by a detection apparatus which is configured for the simultaneous detection of the fluorescence radiation excited by the light sheets that are arranged parallel to one another.

Abstract: The invention relates to a light sheet microscope which comprises an illumination apparatus, which generates coherent illumination light for several illumination wavelengths, a beam-shaping module (8) for generating a light sheet from illumination light, an illumination objective (10) for the illumination of a specimen (1) with the light sheet and a detection objective (12) for imaging light which is emitted by the specimen onto a laminar detector, wherein the optical axis of the detection objective (12) forms an angle different from 0° and 180° with the optical axis of the illumination objective (10).

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: The invention relates to a device and a method for imaging a sample (2) arranged in an object plane (1). Such a device comprises an optical relay system (3) which images an area of the sample (2) from the object plane (1) into an intermediate image plane (4). Here, the object plane (1) and the intermediate image plane (4) with an optical axis (5) of the relay system (3) include an angle different from 90°. The optical relay system (3) is composed of several lenses. The device also comprises an optical imaging system (6) with an objective, the optical axis (7) of which lies perpendicularly on the intermediate image plane (4) and which is focused on the intermediate image plane (4), with the result that the object plane (1) can be imaged undistorted onto a detector (8).

Abstract: A microscope including an illumination objective with a first optical axis, embodied to produce a light sheet, and a detection objective with a second optical axis, embodied to detect light coming from the specimen plane. The illumination objective and the detection objective are aligned relative to one another and the specimen plane so that the first and second optical axes intersect in the specimen plane and include a substantially right angle therebetween. The optical axes each include an angle which differs from zero with a reference axis directed orthogonal to the specimen plane. An overview illumination apparatus for wide-field illumination of the specimen plane, includes an illumination optical unit with a third optical axis. The characterizing feature is that the detection objective is provided to detect both light from the light sheet and light from the illumination optical unit. A method is also provided for operating a light sheet microscope.

Abstract: The invention relates to a method for determining height information of a sample, and to a scanning microscope. The method comprises the following steps: generating an illumination spot; illuminating the sample with the illumination spot; capturing an image of a reflection of the illumination spot at the sample; evaluating the lateral distribution of the image; determining the height information from the lateral distribution; wherein the illumination spot has a three dimensional illumination pattern and/or the image in a detection beam path has a three dimensional detection pattern. The scanning microscope is characterized in that an illumination device (07) and/or a detector device comprise(s) a means for generating a three dimensional pattern with a change in the lateral intensity distribution that is asymmetrical along the optical axis, and an evaluation device is configured to determine height information from the detector signal.

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 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 method for the three-dimensional imaging of a sample in which image information from different depth planes of the sample is stored in a spatially resolved manner, and the three-dimensional image of the sample is subsequently reconstructed from this stored image information is provided. A reference structure is applied to the illumination light, at least one fluorescing reference object is positioned next to or in the sample, images of the reference structure of the illumination light, of the reference object are recorded from at least one detection direction and evaluated. The light sheet is brought into an optimal position based on the results and image information of the reference object and of the sample from a plurality of detection directions is stored. Transformation operators are obtained on the basis of the stored image information and the reconstruction of the three-dimensional image of the is based on these transformation operators.