Abberior Instruments Gmbh

Fondazione Istituto Italiano Di Tecnologia

Benjamin Harke

Lars Kastrup

Matthias Reuss

Christian Wurm

Gerald Donnert

Andreas Schoenle

Roman Schmidt

Alberto Diaspro

Winfried Willemer

Giuseppe Vicidomini

Andreas Schönle

Joern Heine

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

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

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 methods of high-resolution imaging a structure of a sample, the structure being marked with fluorescence markers, the sample is subjected to a light intensity distribution including an intensity maximum of focused fluorescence excitation light to selectively scan partial areas of interest of the sample. Fluorescence light emitted out of the sample is registered and allocated to a respective location of the light intensity distribution in the sample. The subjection of the sample to at least one part of the light intensity distribution is terminated at each location of the light intensity distribution, if at least one criterion of the following criteria is met: (a) a predetermined maximum light amount of the fluorescence light emitted out of the sample has been registered, and (b) a predetermined minimum light amount of the fluorescence light emitted out of the sample has not been registered within a predetermined period of time.

Methods of high-resolution imaging a structure of a sample, the structure being marked with fluorescence markers

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

Method of optical microscopy by scanning a sample containing an excitable species, the method comprising:

Stimulated emission-depletion (STED) microscopy based on time gating of excitation beam and synchronous detection of fluorescence emission

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.

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Goettingen, Germany

Benjamin Harke