The invention discloses a programmable annular LED illumination-based high efficiency quantitative phase microscopy imaging method, the proposed method comprising the following steps: the derivation of system optical transfer function in a partially coherent illumination imaging system; the derivation of phase transfer function with the weak object approximation under the illumination of tilted axially symmetric coherent point illumination source; the extension of illumination from an axially symmetric coherence point source to a discrete annular point source, and the optical transfer function can be treated as an incoherent superposition of each pair of tilted axially symmetric coherent point sources. The acquisition of raw intensity dataset; the implementation of deconvolution for quantitative phase reconstruction. The invention derives the system phase transfer function under the tilted axially symmetric point light source in the case of partially coherent illumination, and promotes the optical phase transfer function of the discrete annular point light source. The programmability characteristic of LED array enables the annular illumination aperture to be flexibly adjustable, being applicable to different microscopic objects with different numerical apertures, and improving the compatibility and flexibility of the system.

A system, including an optical imaging assembly configured to image a sample at an object plane to an image plane; an image sensor arranged at the image plane and configured to capture images of the sample for a field of view of the system; a light source configured to emit light having a wavelength, λ; a spatial light modulator (SLM) arranged to receive the light emitted from the light source and to provide a spatially modulated light pattern; one or more optical elements arranged to receive the spatially modulated light pattern from the SLM and to direct the spatially modulated light pattern to the image plane; and an electronic controller in communication with the image sensor and the spatial light modulator, the electronic controller being programmed to identify one or more targets in the field of view of the optical imaging assembly and to control the spatial light modulator to selectively direct light from the light source to the one or more targets identified by the electronic controller.

There is provided a measurement apparatus including a control unit configured to control an on/off state of illumination that does not contribute to acquisition of measurement data on the basis of an acquisition time period of the measurement data.

An in-line holographic microscope can be used to analyze on a frame-by-frame basis a video stream to track individual colloidal particles' three-dimensional motions. The system and method can provide real time nanometer resolution, and simultaneously measure particle sizes and refractive indexes. Through a combination of applying a combination of Lorenz-Mie analysis with selected hardware and software methods, this analysis can be carried out in near real time. An efficient particle identification methodology automates initial position estimation with sufficient accuracy to enable unattended holographic tracking and characterization.

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.

A microscope apparatus comprises: an illumination optical system that guides light from a light source to a sample; a detection unit that detects light from the sample; a detection optical system that has an objective lens and guides light from the sample to the detection unit; a mask that allows a portion of light from the sample and light from the light source to pass therethrough, and blocks the other portion; a mask-switching unit that changes mask patterns of the mask; a microscope control unit; and an information-processing device. The microscope control unit controls the mask-switching unit to change mask patterns. The information-processing device obtains information about the amount of movement of the focus position of the optical system including the objective lens when mask patterns are changed, and calculates the refractive index of the sample based on the obtained information about the amount of movement of the focus position.

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 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.