Abstract: The present specification relates to a method for light microscopic examination of a sample (6), in particular by means of laser scanning or MINFLUX microscopy, in which a drift of the sample (6) or of an object in a sample (6) with respect to the light microscope (26) is detected and, if necessary, corrected. In particular, the present specification relates to a corresponding method for examining the sample (6) using laser scanning or MINFLUX microscopy. For this purpose, reference markers (8, 13) are located in the sample, the position of which is repeatedly determined according to the MINFLUX principle in order to determine the drift.

Abstract: The invention relates to a detection device (2) for a laser scanning microscope, the detection device (2) having a light inlet (4), at least one filter module (14) and at least one spatially resolving detector (22) and being configured to guide light from the light inlet (4) to the filter module (14) and from there to the spatially resolving detector (22), at least one filter module (14) being designed as a continuous filter module with two continuously tunable filter elements (16), and at least one compensator element (26) being arranged optically downstream of the continuous filter module (14), by means of which a focal position of light on the spatially resolving detector (22) can be adjusted.

Abstract: A fluorescence microscope (10) includes a sample illumination beam path including a source (9) for illumination light, a first wave front modulator (24) for providing the focused illumination light (8) with a central intensity minimum, a beam splitter (26) and a second adjustable wave front modulator (34) arranged in a pupil plane (30) of an objective (20). A first detection beam path section including the second wave front modulator (34) and a telescope (11) and ending at the beam splitter (26) coincides with the sample illumination beam path. A separate second detection beam path section includes a detector (38) for luminescence light from a sample. The telescope (11) images a first pupil (31) formed in the pupil plane (30) in a smaller second pupil (32), and transfers a beam of the illumination light (8) collimated in the second pupil (32) into an expanded beam collimated in the first pupil (31).

Abstract: An apparatus for selectively shaping phase fronts of a first light beam that is incident along an optical axis and that has a first linear input polarization direction running orthogonal to the optical axis comprises birefringent optical material arranged in all or all bar one of at least three different partial areas that follow to one another in a direction around the optical axis. The optical material is arranged such that a phase of the first light beam is delayed differently in the different partial areas to an extent that increases from partial area to partial area over a round around the optical axis, whereas a phase of a second light beam that is incident along the optical axis and that has a second linear input polarization direction orthogonal to the first linear input polarization direction and to the optical axis is not delayed differently in the different partial areas.

Abstract: In a first step of a method of microscopically recording images of samples extending in three dimensions, a first sectional image that is parallel to an optical axis of a microscope objective lens is recorded by scanning a sample with a focused excitation light distribution in a sectional area parallel to the optical axis of the microscope objective, wherein the excitation light distribution is corrected by a correction device according to initial adjustment values for adjustment parameters of an aberration correction function. In a second step, the first sectional image is evaluated. In a third step, new adjustment values for the adjustment parameters are defined. In a fourth step, further image data are recorded by scanning the sample with the focused excitation light distributions, wherein the excitation light distribution is corrected by the correction device according to the new adjustment values for the adjustment parameters of the aberration correction function.

Abstract: For setting a laser-scanning fluorescence microscope to a correct alignment in which an intensity maximum of excitation light and an intensity minimum of fluorescence inhibition light coincide in a focal area of an objective lens, a structure in a sample marked with a fluorescent dye is scanned with the intensity maximum of the excitation light to generate first and second pictures of the sample, the first picture corresponding to a higher and the second picture corresponding to a lower intensity of the fluorescence inhibition light. A spatial offset of a first image of the structure in the first picture with regard to a second image of the structure in the second picture is calculated; and the intensity maximum of the excitation light is shifted with regard to the intensity minimum of the fluorescence inhibition light in the direction of the offset calculated to set the microscope to the correct alignment.

Abstract: A fluorescence microscope (10) comprises a sample illumination beam path including a source (9) for illumination light, a first wave front modulator (24) for providing the focused illumination light (8) with a central intensity minimum, a beam splitter (26) and a second adjustable wave front modulator (34) arranged in a pupil plane (30) of an objective (20). A first detection beam path section including the second wave front modulator (34) and a telescope (11) and ending at the beam splitter (26) coincides with the sample illumination beam path. A separate second detection beam path section includes a detector (38) for luminescence light from a sample. The telescope (11) images a first pupil (31) formed in the pupil plane (30) in a smaller second pupil (32), and transfers a beam of the illumination light (8) collimated in the second pupil (32) into an expanded beam collimated in the first pupil (31).

Abstract: An apparatus for selectively shaping phase fronts of a first light beam that is incident along an optical axis and that has a first linear input polarization direction running orthogonal to the optical axis comprises birefringent optical material arranged in all or all bar one of at least three different partial areas that follow to one another in a direction around the optical axis. The optical material is arranged such that a phase of the first light beam is delayed differently in the different partial areas to an extent that increases from partial area to partial area over a round around the optical axis, whereas a phase of a second light beam that is incident along the optical axis and that has a second linear input polarization direction orthogonal to the first linear input polarization direction and to the optical axis is not delayed differently in the different partial areas.

Abstract: A high resolution laser scanning microscope has beam shaping elements configured to shape a beam of fluorescence inhibiting light which is directed into a back aperture of an objective connected to form an intensity minimum delimited by intensity maxima of the fluorescence inhibiting light in a focus of the objective. A plurality of optical elements including the objective and the beam shaping elements are arranged in a beam path of the beam to the focus. Using the microscope includes removing or exchanging or altering or adding at least one of the optical elements arranged in the beam path of the beam of fluorescence inhibiting light, and compensating a variation of polarization varying properties of the plurality of the optical elements, that is caused by removing or exchanging or altering or adding the at least one optical element, by adapting the beam shaping elements to the variation.

Abstract: For setting a laser-scanning fluorescence microscope to a correct alignment in which an intensity maximum of excitation light and an intensity minimum of fluorescence inhibition light coincide in a focal area of an objective lens, a structure in a sample marked with a fluorescent dye is scanned with the intensity maximum of the excitation light to generate first and second pictures of the sample, the first picture corresponding to a higher and the second picture corresponding to a lower intensity of the fluorescence inhibition light. A spatial offset of a first image of the structure in the first picture with regard to a second image of the structure in the second picture is calculated; and the intensity maximum of the excitation light is shifted with regard to the intensity minimum of the fluorescence inhibition light in the direction of the offset calculated to set the microscope to the correct alignment.

Abstract: A high resolution laser scanning microscope has beam shaping elements configured to shape a beam of fluorescence inhibiting light which is directed into a back aperture of an objective connected to form an intensity minimum delimited by intensity maxima of the fluorescence inhibiting light in a focus of the objective. A plurality of optical elements including the objective and the beam shaping elements are arranged in a beam path of the beam to the focus. Using the microscope includes removing or exchanging or altering or adding at least one of the optical elements arranged in the beam path of the beam of fluorescence inhibiting light, and compensating a variation of polarization varying properties of the plurality of the optical elements, that is caused by removing or exchanging or altering or adding the at least one optical element, by adapting the beam shaping elements to the variation.

Abstract: A scanner head for high-resolution scanning fluorescence microscopy comprises a first connector to connect the scanner head to a light microscope, second connectors to connect the scanner head to a light source and a fluorescence light detector, a beam shaper to shape a first part of the light from the light source into a first light intensity distribution in the focus of the light microscope comprising an intensity minimum surrounded by intensity maxima and a second part of the light into a second light intensity distribution in the focus of the light microscope comprising an intensity maximum at the location of the intensity minimum of the first light intensity distribution four tilting mirrors configured scan a sample with the light beam, and a deflector to deflect the fluorescence light to the second optical waveguide port.

Abstract: A device comprises two polarization-selective optical elements for separately modulating wave fronts of two components of a collimated light beam, which are transversally polarized in orthogonal directions. The two polarization-selective optical elements are first and second partial areas of one spatial light modulator (SLM) diffracting the light beam in backward direction. A mirror arranged between the first and second partial areas of the SLM reflects the light beam coming from the first partial area towards the second partial area. A wave plate arranged between the first partial area and the second partial area of the SLM rotates the polarization directions of both components of the light beam by 90°. The mirror reflects the first and second components of the light beam as parallel bundles of light rays resulting in a lateral offset between the first and second components of the light beam behind the second partial area of the SLM.

Abstract: A device comprises two polarization-selective optical elements for separately modulating wave fronts of two components of a collimated light beam, which are transversally polarized in orthogonal directions. The two polarization-selective optical elements are first and second partial areas of one spatial light modulator (SLM) diffracting the light beam in backward direction. A mirror arranged between the first and second partial areas of the SLM reflects the light beam coming from the first partial area towards the second partial area. A wave plate arranged between the first partial area and the second partial area of the SLM rotates the polarization directions of both components of the light beam by 90°. The mirror reflects the first and second components of the light beam as parallel bundles of light rays resulting in a lateral offset between the first and second components of the light beam behind the second partial area of the SLM.

Abstract: A scanner head for high-resolution scanning fluorescence microscopy comprises a first connector to connect the scanner head to a light microscope, second connectors to connect the scanner head to a light source and a fluorescence light detector, a beam shaper to shape a first part of the light from the light source into a first light intensity distribution in the focus of the light microscope comprising an intensity minimum surrounded by intensity maxima and a second part of the light into a second light intensity distribution in the focus of the light microscope comprising an intensity maximum at the location of the intensity minimum of the first light intensity distribution four tilting mirrors configured scan a sample with the light beam, and a deflector to deflect the fluorescence light to the second optical waveguide port.

Abstract: In a STED fluorescence light microscope pulses of excitation light (3) are applied to a sample, which excite fluorescent entities contained in the sample for fluorescence, and which are focused on at least one focal area. Further, de-excitation light (12) is applied to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave. Fluorescence light emitted by the excited fluorescent entities in the sample is registered after each pulse of the excitation light (3) and overlapping with applying the de-excitation light (13) with high temporal resolution between consecutive pulses of the excitation light (3).

Abstract: A fluorescence light scanning microscope (2) comprises a light source providing excitation light (8) for exciting a fluorophore in a sample to be imaged for spontaneous emission of fluorescence light, and suppression light (7) for suppressing spontaneous emission of fluorescence light by the fluorophore on a common optical axis (4), the suppression wavelength differing from the excitation wavelength; an objective (19) focusing both the excitation (8) and the suppression (7) light to a focus point; a detector (21) detecting fluorescence light (11) spontaneously emitted by the fluorophore; and a chromatic beam shaping device (1) arranged on the common optical axis (4), and including a birefringent chromatic optical element (3) adapted to shape a polarization distribution of the suppression light (7) such as to produce an intensity zero at the focus point, and to leave the excitation light such as to produce a maximum at the focus point.

Abstract: In a STED fluorescence light microscope pulses of excitation light (3) are applied to a sample, which excite fluorescent entities contained in the sample for fluorescence, and which are focused on at least one focal area. Further, de-excitation light (12) is applied to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave. Fluorescence light emitted by the excited fluorescent entities in the sample is registered after each pulse of the excitation light (3) and overlapping with applying the de-excitation light (13) with high temporal resolution between consecutive pulses of the excitation light (3).

Abstract: A fluorescence light scanning microscope (2) comprises a light source providing excitation light (8) for exciting a fluorophore in a sample to be imaged for spontaneous emission of fluorescence light, and suppression light (7) for suppressing spontaneous emission of fluorescence light by the fluorophore on a common optical axis (4), the suppression wavelength differing from the excitation wavelength; an objective (19) focusing both the excitation (8) and the suppression (7) light to a focus point; a detector (21) detecting fluorescence light (11) spontaneously emitted by the fluorophore; and a chromatic beam shaping device (1) arranged on the common optical axis (4), and including a birefringent chromatic optical element (3) adapted to shape a polarization distribution of the suppression light (7) such as to produce an intensity zero at the focus point, and to leave the excitation light such as to produce a maximum at the focus point.

Abstract: An oscillating motor includes a cylinder which is filled with hydraulic medium and which has at least one rib on its inside wall, whereby a motor shaft with at least one vane is supported inside the cylinder with freedom to oscillate, a sleeve concentric to the motor shaft, and a pressure-preloaded sealing arrangement inside each of at least two ring-shaped spaces between the cylinder and the motor shaft, which sealing arrangements seal the working chambers formed by the cylinder and its rib, the motor shaft, and the vane together with the cylinder covers at the ends. The ring-shaped spaces are connected to each other by a pressure equalization channel, and an axial groove, which forms the pressure equalization channel, is present in the contact area between the motor shaft and the sleeve.