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.

Configurations are disclosed for a health system to be used in various healthcare applications, e.g., for patient diagnostics, monitoring, and/or therapy. The health system may comprise a light generation module to transmit light or an image to a user, one or more sensors to detect a physiological parameter of the user's body, including their eyes, and processing circuitry to analyze an input received in response to the presented images to determine one or more health conditions or defects.

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 method, an arrangement for microscopy and a microscope for three-dimensional imaging in microscopy, in which aberrations of a specimen detection radiation coming from a specimen are corrected in a detection beam path by means of a correction element and the corrected specimen detection radiation is captured in a spatially resolved form. The inventions are distinguished by the fact that a best-possible correction setting of the correction element, with which aberrations occurring at the time are reduced as much as possible, is determined; and, on the basis of the best-possible correction setting, a flawed correction setting is determined, a setting with which aberrations occurring lead to an asymmetric point spread function of the specimen detection radiation.

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.

Apparatus and method for capturing an image having a detection beam path for guiding detection radiation from a sample to a detector having a plurality of detector elements. The detector has no more than ten and, preferably, four or five detector elements; and an evaluation unit, which is configured to carry out an evaluation in accordance with the Airyscan method on the image data captured by means of the detector and which generates a high-resolution image.

Systems and methods for passive multi-directional illumination in light-sheet fluorescence imaging and microscopy are disclosed herein. An elliptical light shaping diffuser is placed in the illumination path between the source of a light-sheet and the illuminated sample. The light-sheet is diffused anisotropically along two directions perpendicular to its propagation direction, eliminating stripe artifacts in obtained images. The method includes converting a light-sheet into an elliptically diffuse light-sheet by passing it through an elliptical light shaping diffuser, illuminating a sample with the elliptically diffuse light-sheet. The system includes a light-sheet source, an elliptical light shaping diffuser adapted to convert the light-sheet into an elliptically diffuse light-sheet to illuminate the sample, typical microscopy optics and lenses, and image capturing elements.

A system for illuminating a microscopy specimen includes an illumination source configured to emit a light that travels along an illumination path to illuminate the microscopy specimen placed on an optical detection path of an optical microscope. The system also includes optical elements in the illumination path and configured to at least in part transform the light from the illumination source into a light sheet illuminating the microscopy specimen. The optical elements include an electronically tunable lens configured to vary a focal distance of the electronically tunable lens to dynamically vary a position of a waist of the light sheet illuminating the microscopy specimen. The optical elements include a deflector configured to vertically move the light sheet to illuminate the microscopy specimen at different horizontal planes.

Some implementations of the disclosure relate to an imaging system, including: a sample holder to support a sample container having multiple sample locations; an optical stage having; an assembly comprising one or more actuators physically coupled to the sample holder to tilt the sample holder relative to the optical stage during imaging of the multiple sample locations to focus the optical stage onto a current sample location; a first light source to project a first pair of spots on the sample container; and a controller to control, based on a sample tilt determined from a first separation measurement of the first pair of spots from one or more images taken by an image sensor at one or more of the sample locations, the one or more actuators to tilt the sample holder along a first direction of the imaging or a second direction substantially perpendicular to the first direction.