In a method for processing microscope images, at least one microscope image is provided as input image for an image processing algorithm. An output image is created from the input image by means of the image processing algorithm. The creation of the output image comprises adding low-frequency components for representing solidity of image structures of the input image to the input image, wherein the low-frequency components at least depend on high-frequency components of these image structures and wherein high-frequency components are defined by a higher spatial frequency than low-frequency components. A corresponding computer program and microscope system are likewise described.

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.

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.

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.

An image display method includes the following operations (a) to (e). The (a) is of obtaining a plurality of two-dimensional images by two-dimensionally imaging a specimen, in which a plurality of objects to be observed are present three-dimensionally in the specimen, at a plurality of mutually different focus positions. The (b) is of obtaining image data representing a three-dimensional shape of the specimen. The (c) is of obtaining a three-dimensional image of the specimen based on the image data. The (d) is of obtaining the two-dimensional image selected from the plurality of two-dimensional images or a two-dimensional image generated to be focused on the plurality of objects based on the plurality of two-dimensional images as an integration two-dimensional image. The (e) is of integrating the integration two-dimensional image obtained in the (d) with the three-dimensional image obtained in the (c) and displaying an integrated image on a display unit.

A method for measuring a position of a fluorophore includes configuring a set of compact light distributions, the set having at least one member, each light distribution characterized by a center, so that there is substantially zero intensity at the center of the set of compact light distributions. The method additionally includes moving the set of compact light distributions in relation to a set of hypothesized positions of the fluorophore, detecting, in a plurality of locations corresponding to the hypothesized set of positions, a set of images; and estimating the position of the fluorophore, by determining from the set of images a set of parameters describing the position of the fluorophore using an inverse problem method.

A computer-implemented method for determining a value for at least three setting parameters by means of an input unit in the form of a graphical user interface with an input cursor positionable in an input area is provided. At least two of the at least three setting parameters can be set independently of one another. The method includes determining a position of the input cursor within the input area; determining a coordinate of the position of the input cursor for each particular setting parameter of the at least three setting parameters as a function of a distance in each case of the position of the input cursor from at least one coordinate origin allocated to the particular setting parameter; and determining a value for each particular setting parameter of the at least three setting parameters as a function of the coordinate determined for each particular setting parameter.

Systems, methods, and apparatus for differential phase contrast microscopy by transobjective differential epi-detection of forward scattered light are provided. In some embodiments, a microscope objective comprises: a housing with mounting threads at a second end; optical components defining an optical axis, comprising: an objective lens mounted at a first end, configured to collect light from a sample placed in a field of view, the plurality of optical components create a pupil plane at a first distance along the optical axis at which rays having the same angle of incidence on the objective lens converge at the same radial distance from the optical axis; a photodetector within the housing offset from the optical axis at a second distance along the optical axis; and another photodetector within the housing at second distance along the optical axis and offset from the optical axis in the opposite direction from the first photodetector.

Laser emission based microscope devices and methods of using such devices for detecting laser emissions from a tissue sample are provided. The scanning microscope has first and second reflection surfaces and a scanning cavity holding a stationary tissue sample with at least one fluorophore/lasing energy responsive species. At least a portion of the scanning cavity corresponds to a high quality factor (Q) Fabry-Pérot resonator cavity. A lasing pump source directs energy at the scanning cavity while a detector receives and detects emissions generated by the fluorophore(s) or lasing energy responsive species. The second reflection surface and/or the lasing pump source are translatable with respect to the stationary tissue sample for generating a two-dimensional scan of the tissue sample. Methods for detecting multiplexed emissions or quantifying one or more biomarkers in a histological tissue sample, for example for detection and diagnosis of cancer, or other disorders/diseases are provided.

An arrangement for increasing resolution of a laser scanning microscope has a simplified adjustment and lower susceptibility to errors. The pupil beam from the laser scanning microscope is coupled into a shortened common path interferometer, to make wavefronts of a pupil image mirrored at at least one axis and wavefronts of an unchanged pupil image interfere. The area of a pupil from the pupil beam is split into two complementary portions P and Q producing two partial beams separately supplied to at least one beam deflection means by total-internal reflection along the common path interferometer. The light of the interferometer branches from transmitted light of the one interferometer branch and reflected light of the other interferometer branch is made to interfere at a partly transmissive beam splitter layer to cause constructive interference C and destructive interference D of the wavefronts from the two different portions P and Q of the pupil.