Abberior Instruments Gmbh

Max-planck-gesellschaft Zur F&#xf6;rderung Der Wissenschaften E. V.

Lars Kastrup

Benjamin Harke

Matthias Reuss

Christian Wurm

Gerald Donnert

Roman Schmidt

Andreas Schoenle

Stefan W Hell

Vladimir Belov

Katrin Willig

Kirill Kolmakov

Johanna Wildanger

Shamil Nizamov

Heiko Schill

Volker Westphal

Joachim Fischer

Matthias Henrich

Winfried Willemer

Andreas Schönle

Joern Heine

Vladimir N Belov

Jörn Heine

Christian A Wurm

The invention is directed to a method for recording a motion trajectory of an individual particle in a sample and to a light microscope performing this method. Starting from an at least approximately known initial position, the particle is scanned with an intensity distribution of a scanning light comprising a local intensity minimum. When illuminated with the scanning light, the particle to be tracked generates a detectable light signal, from the intensity of which updated coordinates of the particle are calculated. According to the invention, the scanning is terminated when a second measured variable detected in parallel satisfies a termination criterion.

Method and microscope for recording trajectories of individual particles in a sample

A method for punctiform illumination of a sample with a MINFLUX microscope has the sample being sequentially illuminated at illumination points of a predefined or predefinable illumination point pattern. The lateral extent of the illumination point pattern is smaller than the longest wavelength of the illumination light. The illumination points are always illuminated exclusively with a time offset. A distinct individual light source is assigned to each illumination point of the illumination point pattern. Each illumination point is illuminated by the focus of an illumination light bundle of the individual light source.

Method and device for illuminating a sample in a microscope in points

The present disclosure is directed to a method of disturbance correction and to a laser scanning microscope carrying out this method. Specifically, it is directed to an image recording method according to the MINFLUX principle, in which a spatially isolated fluorescence dye molecule is illuminated at a sequence of scan positions by an intensity distribution with a local intensity minimum, and the number of fluorescence photons emitted by the fluorescence dye molecule is detected at each of the scan positions. The location of the molecule is determined with a high spatial resolution from the scan positions and the numbers of fluorescence photons. A disturbance is captured when illuminating the fluorescence dye molecule and detecting the fluorescence light, said disturbance being considered in corrective fashion when determining the location of the fluorescence dye molecule.

Method of disturbance correction, and laser scanning microscope having disturbance correction

An apparatus for detecting movements of a sample with respect to an objective includes imaging optics which include the objective, which have an image plane and which are configured to image light from at least one reference object that is connected to the sample arranged in front of the objective into reference object images in the image plane. The apparatus further includes a camera which is arranged in the image plane of the imaging optics and which is configured to record the reference object images at consecutive points in time, and an optical device arranged between the objective and the camera in a plane that is Fourier-conjugated with respect to the image plane. The optical device is configured to mask out low spatial frequencies of reference object images which the imaging optics image into the image plane.

Detecting movements of a sample with respect to an objective

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

Method and light microscope for a high-resolution examination of a sample

A bandpass filter for light has variable lower and upper cut-off wavelengths. The bandpass filter comprises an areal long-pass filter defining the variable lower cut-off wavelength and an areal short-pass filter defining the variable upper cut-off wavelength. The long-pass filter has different lower cut-off wavelengths in different first area regions which follow to one another in a first direction, and the short-pass filter has different upper cut-off wavelengths in different second area regions which follow to one another in a second direction. The long-pass filter and the short-pass filter are connected in series and spatially fixed relative to one another. The first direction and the second direction are oriented crosswise to one another.

Bandpass filter for light having variable lower and upper cut-off wavelengths

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.

Method of aligning a laser-scanning fluorescence microscope and laser-scanning fluorescence microscope having an automatic aligning system

In order to generate rasterized images of a sample, a pixel size of image points of a rasterized image is set and photons emitted out of the sample which were detected, and for each of which a position of an effective local excitation of the sample for emitting the respective detected photon has been recorded are assigned to that image point of the rasterized image into which the position of the effective local excitation recorded for the respective detected photon falls. To set the pixel size of the image points to an optimized pixel size, the positions of the effective local excitation of the sample for emitting the detected photons are evaluated.

Method, device and laser scanning microscope for generating rasterized images

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.

Method of using a high resolution laser scanning microscope and high resolution laser scanning microscope

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&#xb0;. 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.

Device for separately modulating the wave fronts of two components of a light beam and microscope comprising the device

The invention relates to novel fluorescent dyes with phosphorylated hydroxymethyl groups, a method for preparing the same as well as to their use in imaging techniques. The fluorescent dyes are coumarin, rhodamine or BODIPY dyes having of one of the following general formulae I-III: wherein W=OP(O)YYor P(O)YY, where Yand Yindependently denote any of the following residues: OH, O(&#x2212;), ORand OR, NHRand NHR, NRRand NRR, ORand NHR, ORand NRR, NHRand NRR; and any salt thereof.

Fluorescent dyes with phosphorylated hydroxymethyl groups and their use in light microscopy and imaging techniques

In an optical assembly having a light source which provides two optically different light components with essentially planar wavefronts on an optical axis, wherein the light components differ at least in their wavelength; in the case of an objective lens which projects the two optically different light components into a projection space; and in the case of an optical component which is arranged on the optical axis and has an plane through which the wavefronts of the two light components pass and in which at least two different areas of the optical component with different dispersion behaviors n(&#x3bb;) abut against one another in the lateral direction with respect to the optical axis; the optical component causes phase shifts of the wavefronts of the two light components, wherein the phase shift of the wavefronts of the one light component differs by at least one quarter of the wavelength of that light component between the two different areas, and wherein the phase shift of the wavefronts of the other light component does not differ between the two different areas, such that an intensity distribution of the one light component in the projection space by interference with itself differs from an intensity distribution of the other light component in the projection space.

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Goettingen, Germany

Lars Kastrup