A light sheet fluorescence microscope includes a light source configured to emit excitation light suitable for inducing fluorescent light emitted from a specimen, a detector configured to detect the fluorescent light from the specimen, and an optical system configured to illuminate the specimen with a light sheet formed from the excitation light, and to guide the fluorescent light from the illuminated specimen to the detector. The optical system includes an objective facing the specimen, the objective being configured to collect the fluorescent light emitted from the specimen. The light source is further configured to emit manipulation light suitable for photomanipulating the specimen. The optical system is further configured to direct the manipulation light through a spatially limited sub-area of an entrance pupil of the objective onto the specimen along a light propagation direction which is different from a light propagation direction of the light sheet.

Methods and systems for generating non-diffracting light sheets for multicolor fluorescence microscopy are disclosed. A method for generating a non-diffracting light patterned Bessel sheet comprises transmitting an input light beam through a Fourier transform lens the input light beam has a spatial intensity pattern at a first plane, and a Fourier plane is formed after the Fourier transform lens to obtain a first light beam; transmitting the first light beam through an annulus mask to obtain a second light beam; and transmitting the second light beam through an excitation objective lens to form a non-diffracting patterned light sheet. A method for generating a non-diffracting light line Bessel sheet comprises transmitting an input light beam at a first lane through an annulus mask to obtain a first light beam; and transmitting the first light beam through an excitation objective lens to form a non-diffracting Bessel light sheet.

A sample observation device includes: an emission optical system that emits planar light to a sample on an XZ plane; a scanning unit that scans the sample in a Y-axis direction so as to pass through an emission surface of the planar light; an imaging optical system that has an observation axis inclined with respect to the emission surface and forms an image of observation light generated in the sample; an image acquisition unit that acquires a plurality of pieces of XZ image data corresponding to an optical image of the observation light; and an image generation unit 8 that generates XY image data based on the plurality of pieces of XZ image data. The image generation unit extracts an analysis region of the plurality of pieces of XZ image data acquired in the Y-axis direction, integrates brightness values of at least the analysis region in a Z-axis direction to generate X image data, and combines the X image data in the Y-axis direction to generate the XY image data.

A method is presented for obtaining an optically-sectioned image of a sample. The method comprises: providing an illumination beam through an imaging lens such that the illumination beam is focused at a focal plane of the imaging lens; obtaining a plurality of images of the sample. Obtaining comprises providing the illumination beam at a plurality of lateral positions on the focal plane and obtaining each image at each lateral position of the illumination beam, such that an intensity of the illumination beam on a portion of the sample at the focal plane varies for each of the plurality of lateral positions. The method further comprises detecting, using a detector, signals collected via the imaging lens; and constructing the optically-sectioned image based on the plurality of images. The constructing comprises: obtaining a plurality of signal values from the portion of the sample from the plurality of images; evaluating a threshold for the portion; and evaluating a pixel value by integrating a fraction of the plurality of signal values based on the threshold.

A single-particle localization microscope, including an optical system configured to illuminate a sample region with a sequence of light patterns having spatially different distributions of illumination light adapted to cause a single particle located in the sample region to emit detection light, a detector configured to detect a sequence of intensities of the detection light emerging from the sample region in response to the sequence of illuminating light patterns, and a processor configured to determine, based on the sequence of intensities of the detection light, an arrangement of potential positions for locating the particle. The processor further illuminates the sample region with at least one subsequent light pattern, causes detection of at least one subsequent intensity, and decides, based on the at least one subsequent intensity of the detection light, which one of the multiple potential positions represents an actual position of the particle in the sample region.

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.

A SPIM-microscope (Selective Plane Imaging Microscopy) and a method of operating the same having a y-direction illumination light source and a z-direction detection light camera. An x-scanner generates a sequential light sheet by scanning the illumination light beam in the x-direction. An electronic zoom is provided that is adapted to change the scanning length in the x-direction independently of a focal length of the illumination light beam and a size of the light sheet in the y-direction and in the z-direction, wherein the number of image pixels in x-direction is maintained unchanged by the electronic zoom independently of the scanning length in x-direction that has been selected.

A super resolution technique, intended mainly for fluorescence microscopy, acquires the three-dimensional position of an emitter, through a hybrid method, including a number of steps. In a first step the two-dimensional position of an emitter is acquired, using a technique, named in this application as an Abbe's loophole technique. In this technique a doughnut, or a combination of distributions, having a zero intensity at the combined center of the distributions, is projected onto the sample containing the emitter, under conditions wherein the doughnut null is moved towards the emitter to reach a position in which the emitter does not emit light. In a second step, an axial measurement is obtained using a 3D shaping method, characterized by the fact that the emitted light is shaped by an additional optical module creating a shape of the light emitted by the emitter, this shape being dependent of the axial position and means to retrieve the axial position from the shape.

The present invention relates to a method of reading out information from a data carrier and to a data carrier utilizing the concept of structured-illumination microscopy or saturated structured-illumination microscopy.

Provided is a light detection device having a laser light source, a splitting unit, a first modulation unit, a second modulation unit, a first detection unit and a second detection unit that detect light, and a control unit.