42 results about "Light field microscopy" patented technology

A light-field microscopy system includes a light-field microscope, and an image processing system configured to receive images from the light-field microscope and perform a hybrid point-spread function (PSF) operation using a hybrid point-spread function that comprises numerical profiles that are adjusted by experimental positions acquired from an experimentally recorded PSF.

The invention provides a light field microscope three-dimensional image reconstruction system and method integrated with an event camera, and belongs to the technical field of optics. According to the invention, original light field data is synthesized into event data for training a network, the event data is input into an event image reconstruction network module capable of converting continuous event streams into a series of intensity images to obtain light field event images, and then three-dimensional reconstruction of a target area is obtained through a three-dimensional image reconstruction network module. And a new technical reference is provided for solving the problem of how to improve the three-dimensional reconstruction quality and resolution of the light field microscope.

A modality for compressed light field measurement to capture 3D fluorescent signals at kilohertz volume rate, is described. This modality, termed Squeezed Light field Microscopy (SQLFM), acquires the light field with an array of rotated 2D sub-aperture images. An anamorphic relay system applies anisotropic scaling, effectively "squeezing" the image along one axis. This allows the camera sensor to detect the light field with a letterbox-shaped region of interest (ROI). Each squeezed sub-aperture image is complementary to the others, facilitating successful 3D reconstruction from compressed measurement while preserving the nominal FOV and spatial resolution. SQLFM can achieve over a fivefold increase in acquisition rate compared to traditional LFM.

A device for light field microscopy, comprising: a two-dimensionally spatially resolving detector for detecting light radiated by a sample; a detection beam path having at least one microscope objective lens; and at least one multi-lens array for imaging the light radiated by the sample onto the detector, wherein the multi-lens array is arranged in a plane which is optically conjugated with respect to a rear focal plane of the microscope objective lens, or in the vicinity of such a plane; and a control and evaluation unit for activating the detector and for evaluating the measurement data from the detector. The device also includes an adjustable relay optical system in order to image the rear focal plane of the microscope objective lens into the plane in which the multi-lens array is arranged, or in the vicinity of this plane. A method for light field microscopy is also provided.

The present invention discloses a parallax-based automatic focusing and locking method for a light field microscope system, wherein the light field microscope system is used to shoot at a position of a sample defocus distance to obtain four-dimensional phase space data, and the parallax of each perspective of the four-dimensional phase space data relative to the central perspective is calculated using cross-correlation, and a two-dimensional defocus coefficient matrix of the light field microscope system is obtained based on the parallax and defocus distance calculation; the light field microscope system is used to shoot the sample at the current shooting distance to obtain the current two-dimensional parallax matrix of the sample; the two-dimensional parallax matrix S and the two-dimensional defocus coefficient matrix K are linearly fitted to obtain the current defocus amount of the sample; the current shooting distance is adjusted in real time according to the defocus amount until focusing and / or locking is completed. By analyzing the parallax characteristics of the light field image, the present invention can realize the rapid automatic focusing of the light field microscope system and the locking function during the long-term shooting process.

The invention discloses a method for performing three-dimensional positioning by using a light field microscope, which belongs to the field of light field microscope imaging, and comprises the following steps of: S1, performing light field imaging of an object by using the light field microscope, calibrating an observed original light field image and decoding the original light field image into a biplane parameterized four-dimensional format, therefore, a real polar plane image, namely a real EPI, is formed; s2, wave light forward direction models of spherical volumes located at different depths are used for simulating a set of light fields, a depth perception dictionary is constructed, and each atom is a synthetic polar plane image related to the specific depth, namely a synthetic EPI; s3, implementing three-dimensional positioning by executing a convolutional sparse coding model based on a depth perception dictionary and a dry friction inertial near-end gradient algorithm (IPGDF) on the observed polar plane image; according to the invention, the positioning precision and the positioning speed of three-dimensional positioning under the light field microscope are improved.

The invention provides a visual field depth expanding method based on sparse scanning of a light field microscope, which comprises the following steps of: performing sparse scanning on a sample in X, Y and Z axial directions by utilizing a micro-lens array light field imaging system, enabling the overlapping rate between adjacent visual fields to be lower than a preset value, and acquiring multi-visual-field two-dimensional original light field data; performing pixel rearrangement and deconvolution three-dimensional reconstruction on the two-dimensional original light field image of each view to obtain a reconstructed three-dimensional body of each view; calculating the theoretical overlapping region width in the XYZ axial direction according to the sparse scanning step length, and carrying out consistency calibration on the overlapping region in the XYZ axial direction of the adjacent visual fields; and carrying out recursive fusion on the calibrated reconstructed three-dimensional body along the X, Y and Z axial directions to obtain a full-view-field and full-depth-of-view three-dimensional body image. According to the invention, not only can a larger imaging depth range be provided, but also the image definition at different depth levels can be ensured, and the inherent depth-of-field limitation of a traditional optical system is overcome.

The invention discloses a large-depth-of-field snapshot type light field microscope and a preparation method thereof, and belongs to the technical field of photoelectric devices and microscopic imaging, and the preparation method comprises the following steps: designing a large-depth-of-field curved surface micro lens array; performing femtosecond laser direct writing preparation on the curved surface micro lens array; assembling and integrating the curved micro-lens array; preparation and image calibration of a snapshot light field microscope; according to the method, plane configuration is replaced by the curved-surface micro-lens array to enhance parallax, a traditional spherical lens is replaced by a specially designed lens surface type to expand the focal depth, the problem of baseline limitation of a traditional plane micro-lens array is fundamentally solved, the imaging depth of field and the depth perception capability of the optical microscope are greatly improved, and the imaging precision of the optical microscope is improved. And the imaging resolution of the diffraction limit level is reserved. The high-precision preparation of the special function surface type curved surface microlens array provided by the invention can be realized by utilizing a femtosecond laser additive manufacturing technology, and the preparation of the large-field-depth snapshot type light field microscope is finally realized by integrating a customized mechanical adapter and a commercial objective lens.

The invention discloses an automatic Gaussian light sheet center positioning method based on a confocal scanning light field microscope, and relates to the technical field of optical imaging, and the method comprises the steps: setting an initial offset center of a line scanning module based on a target sample, and adjusting the confocal scanning light field microscope; shooting a two-dimensional light field image of the target sample through the adjusted confocal scanning light field microscope, and performing two-dimensional slicing on the two-dimensional light field image according to a preset interval to obtain sub-view-angle images; on the basis of the sub-view-angle images, full-view-field longitudinal energy distribution is obtained, and the full-view-field longitudinal energy distribution is corrected; and calibrating the initial offset center based on the full-view-field longitudinal energy distribution before and after correction. Compared with manual calibration, parameters do not need to be adjusted manually, and automatic Gaussian line light center position positioning and calibration of different systems and different samples are achieved.

The present invention discloses a Fourier light field microscope super-resolution imaging method and system. The method comprises the following steps: (1) inputting a single multi-view light field image with a low angular sampling rate obtained by a Fourier light field microscope into an angle difference network to obtain multiple single-view light field images with a high angular sampling rate; (2) reconstructing the multiple single-view light field images with a high angular sampling rate obtained in step (1) into three-dimensional images on a perspective basis, and superimposing them in the same order as the multiple single-view images in the training sample sampling data of the angle difference network to obtain the Fourier light field microscope super-resolution imaging. The present invention provides a Fourier light field microscope super-resolution imaging method, which interpolates more perspective images in the actual captured light field image through a deep learning algorithm without generating crosstalk, and simultaneously obtains a light field PSF that matches the light field image, and then uses a three-dimensional reconstruction algorithm to obtain a high-quality reconstruction result.

A method for evaluating measurement data from a light field microscope, and an apparatus for light field microscopy wherein the following steps are carried out: a) at least one sample position to be analyzed is selected in a sample; b) images of the sample, which each contain a set of partial images, are recorded at a sequence of recording times using a light field microscope; c) the positions corresponding to the selected sample positions are determined in the partial images of the images recorded in step b); d) the image signals are extracted from at least some of the partial images at the positions determined in partial step c); e) an integrated signal is obtained for a certain recording time by virtue of the image signals extracted for a certain position from partial images of this recording time in step d) being integrated to form the integrated signal; and f) a time profile of the integrated signal is obtained by virtue of step e) being carried out for a plurality of recording times.

This efficiently provides a wide range of transfer functions for light-field microscopy. [Solution] A sample volume to be imaged with multiple sub-images (1~12xy) from different capture angles (Φ) is captured, and for the sub-images (1~12xy) intended for 3D reconstruction, the respective point image distribution function (PSF) is identified. The sub-images (1~12xy) are examined for optical symmetry, the symmetry is captured, and stored along with a calculation rule. The calculation rule can start with the point image distribution function (PSF) and / or processed point image distribution function of the first sub-image (1~12) and generate the corresponding point image distribution function (PSF) and / or processed point image distribution function of further sub-images (2x~12xy) that are symmetric with respect to the first sub-image (1~12), and the point image distribution function (PSF) or processed point image distribution function of further sub-images (2x~12xy) is generated only when necessary.

The invention relates to a method for providing a transfer function in light field microscopy, in which a sample volume to be imaged is determined using a plurality of sub-images; determining a corresponding point image transfer function for the sub-image, and distributing the point image transfer function to the sub-image; checking the symmetry of the basic optical technical data of the sub-image; the confirmed symmetry is detected and saved along with a calculation rule that allows a point image transfer function and / or a processed point image transfer function of a further sub-image symmetrical to the first sub-image to be generated from the point image transfer function and / or the processed point image transfer function of the first sub-image. The invention also relates to a reconstruction method for virtually reconstructing a sample volume detected using a plurality of sub-images from different detection angles in light field microscopy.

A lightfield imaging system is described herein. The lightfield imaging system includes a display device for displaying three-dimensional (3D) video images of a volume of interest and a lightfield microscope assembly. The lightfield microscope assembly includes a microscope housing, a Multiple Angle Capture (MAC) sensor array including a plurality of sensors mounted to the microscope housing, an objective lens assembly mounted to the microscope housing and configured to direct light rays from a volume of interest to each of the sensors, and a microscope controller coupled to the display device and the sensors. The microscope controller includes a memory device for storing computer-executable instructions thereon and one or more processors for executing the computer-executable instructions for performing an algorithm for rendering 3D video images of the volume of interest onto the display device.

The invention discloses a focusing light field microscopic imaging system for a three-dimensional microscopic particle image velocity measurement system and F Compared with the non-focusing light field microscopy system, the focusing light field microscopy system proposed in the present invention significantly improves the spatial resolution of the area near the focal plane of the system objective lens by changing the relative positions of the microlens array and the CCD sensor and the microscope tube lens, and analyzes and proposes a focusing light field microscopy system based on the structural characteristics of the focusing light field camera and the infinity-corrected microscopy system. F Number matching method; The system and method provided by the present invention overcome the problems of velocity vector measurement error and poor resolution of neighboring focal plane areas introduced by aberrations caused by the measuring body being far away from the focal plane of the objective lens in a three-dimensional microscopic particle image velocimetry system based on non-focusing light field imaging, and provide a technical basis for the assembly and debugging of a focusing light field microscopic imaging system used in a three-dimensional microscopic particle image velocimetry system.

The invention discloses a large-view-field light field microscopic image distortion correction method and system, and belongs to the technical field of computer vision, and the method comprises the steps: obtaining a calibration image, and extracting a center coordinate of a central micro lens as a feature point; constructing an ideal circle center grid and an actual circle center grid according to the feature points, and obtaining grid coordinates; the two-dimensional nonlinear distortion model comprises an external parameter matrix, an internal parameter matrix, radial distortion and tangential distortion, and elements in the external parameter matrix comprise image rotation angle parameters and translation parameters; elements in the internal reference matrix comprise scaling parameters and optical center position parameters; substituting the normalized ideal circle center grid coordinates and the actual circle center grid coordinates into the two-dimensional nonlinear distortion model, and solving to obtain distortion parameters of the light field microscope; and performing correction and remapping based on the obtained distortion parameters. According to the method and the system, the distortion correction precision of the large-view-field light field microscopic image is improved.

An apparatus and method for light field microscopy. The apparatus has a light source for emitting excitation light, an excitation beam path for guiding the excitation light onto and into a sample, a two-dimensionally spatially resolving detector for detecting emission light emitted by the sample as a consequence of the irradiation by the excitation light, and a detection beam path having a microscope objective and a multi-lens array for guiding the emission light onto the two-dimensionally spatially resolving detector. The two-dimensionally spatially resolving detector being arranged in the focal plane of the multi-lens array or in a plane optically conjugate thereto, the excitation beam path being configured to illuminate only a portion of the sample in a field of view of the detection beam path with excitation light, with a device, in particular a scanner, being present for variable positioning of the illuminated portion of the sample in the field of view of the detection beam path and with a variable stop device being present. The variable stop device being configured to restrict an effective field of view of the detection beam path on the basis of the position of the illuminated portion in the field of view. The stop device is an electronic stop device and / or the stop device is arranged in an intermediate image plane of the detection beam path upstream of the multi-lens array.

The invention relates to a method for providing an overview image of an object to be imaged in light-field microscopy, in which image data of at least one image of a region of the object is detected by means of a detection optics and a detector; and from the image data, reconstructing a plurality of z-planes of the region to be detected in a computational manner, using a portion of the detected image data for reconstruction, and / or reducing the number of z-planes to be reconstructed. A z-section around and above and below the focal plane is defined along the z-direction. A channel is assigned to each z-section, and image data of the structure of the object detected in the respective z-section is stored in correspondence with the respective channel. According to the invention, a control command is generated by the control device, by means of which the output device is controlled, the structure being output in accordance with the associated channel in each case with a different, however, graphic feature specific to the channel. The invention also relates to a light field microscope for carrying out the method according to the invention.

To automatically three-dimensionally track an object in a microscope system using a light field camera.SOLUTION: A light field three-dimensional tracking control device 110 controls a motor-driven stage 90 movable in XYZ directions to three-dimensionally track an object observed by a light field camera 10 through an objective lens 70 even if the object is in a sample holder 80 mounted on the motor-driven stage 90, and comprises: a light field image analysis unit 30 that analyzes a light field image obtained by imaging the object with the light field camera 10 in a time-continuous manner to track a three-dimensional spatial position of the object in the light field image; and a stage control unit 90 that controls the movement in the XYZ directions of the motor-driven stage 90 so that a relative positional relation between the objective lens 70 and the three-dimensional spatial position of the object tracked by the light field image analysis unit 30 is maintained constant.SELECTED DRAWING: Figure 2

The invention relates to a light-field microscope and a light-field microscopy image capturing method, including the steps of providing an illumination radiation from at least one light source; modulating the illumination radiation by virtue of adjusting the intensity distribution thereof; generating a dynamic light sheet in a sample space, in which a sample to be imaged can be arranged; scanning a region of the sample to be imaged using the light sheet; collecting a detection radiation and detecting image signals using light-field technology. The illumination radiation is modulated by virtue of the light source being controlled and an intensity of the illumination radiation provided by the light source being altered, especially in a controlled fashion over time. Image recordings of the sample are triggered for different positions of the light sheet.

The invention provides a confocal light field microscope, which comprises an objective lens, a dichroic mirror, a microlens array positioned on a conjugate rear pupil plane of the objective lens, a mask positioned on a rear focal plane of the microlens array, a double-sided scanning galvanometer and a reflector group, the double-sided scanning galvanometer comprises a first reflecting surface and a second reflecting surface which are respectively positioned on the front and back surfaces, the first reflecting surface is used for reflecting the exciting light introduced by the dichroic mirror to an imaging object in front of the objective lens, and the reflecting mirror group reflects the excited exciting light to the second reflecting surface so as to be reflected to the photosensitive element through the second reflecting surface; wherein the angle of the double-sided scanning galvanometer is changed, so that the exciting light scans the imaging object through the first reflecting surface and the excited light enters the photosensitive element through the second reflecting surface. According to the confocal light field microscope provided by the invention, the influence of scanning distortion can be eliminated during scanning through a double-sided scanning mechanism, and a series of problems caused by a synchronization error of two times of scanning are avoided.

A method provides an overview image of an object in light field microscopy, in which image data of at least one image of a region of the object are acquired by a detection optical unit and a detector and, on the basis of the image data, a number of z planes (Zn) of the region are computationally reconstructed. Z sections (uhF, F, ohF) around and above and below the focal plane (F) are defined in a z direction and a channel is assigned to each of the z sections (Zn) and image data of structures of the object detected in the respective z sections (Zn) are stored assigned to the respective channel. By a controller, control commands by which an output device is activated are generated, the structures being output according to the respectively assigned channels with different, but channel-specific graphical characteristics. A light field microscope carries out the method.

The application discloses an automatic positioning Gauss light sheet center method based on a confocal scanning light field microscope, and relates to the technical field of optical imaging, and comprises the following steps: based on a target sample, setting an initial offset center of a line scanning module, and adjusting a confocal scanning light field microscope; through the adjusted confocal scanning light field microscope, shooting a two-dimensional light field graph of the target sample, and carrying out two-dimensional slicing on the two-dimensional light field graph according to a preset interval to obtain a sub-view angle graph; based on the sub-view angle graph, obtaining a full-view field longitudinal energy distribution, and correcting the full-view field longitudinal energy distribution; based on the full-view field longitudinal energy distribution before and after correction, calibrating the initial offset center. Compared with manual calibration, the application does not need to manually adjust parameters, and realizes automatic Gauss line light center position positioning and calibration of different systems and different samples.

A method for automatically determining whether to perform denoising includes the following steps: S1: scanning a sample using a light field microscopy system to obtain a light field microscopic image containing 4-dimensional information, where the sample is a sample to be 3D reconstructed; S2: rearranging the light field microscopic image to obtain a multi-view rearranged image; S3: calculating a cleanliness score of the multi-view rearranged image; and determining whether to perform denoising process on the multi-view rearranged image, based on the cleanliness score of the multi-view rearranged image.

The present invention discloses a three-dimensional microscale velocity field measurement method, device, and storage medium. The three-dimensional microscale velocity field measurement method includes: obtaining a particle light field image under light field microscopy imaging; calculating a reconstructed particle distribution of the obtained particle light field image within a measurement volume; obtaining a particle center distribution of the reconstructed particle distribution; calculating a low-resolution three-dimensional displacement field within the measurement volume using a cross-correlation algorithm based on the reconstructed particle distribution; using the calculated low-resolution three-dimensional displacement field as the predicted displacement, performing motion compensation on the obtained particle center distribution to obtain a shifted particle center distribution; and performing cross-validation matching on the obtained shifted particle center distribution to obtain a high-resolution three-dimensional microscale velocity field. Based on the low-resolution three-dimensional displacement field obtained by the cross-correlation algorithm, the present invention uses a cross-validation matching algorithm to match two frames of particles. This method can accurately track individual particles at high concentrations and effectively improve the resolution of the three-dimensional microscale velocity field.

This invention provides a light field microscopy imaging system based on rare-earth upconversion materials, comprising: a first light source, a second light source, a third light source, a spatial filtering module, a first dichroic mirror, a second dichroic mirror, a third dichroic mirror, an aperture, a multi-channel filtering module, an objective lens, a tube lens, a microlens array, and a sensor. Light of different wavelengths emitted from the first, second, and third light sources is transmitted through the spatial filtering module to the corresponding first, second, and third dichroic mirrors. The first, second, and third dichroic mirrors are spatially aligned and beam-combined through the aperture. The combined three-color light sources are focused into the objective lens to achieve wide-field illumination. The rare-earth upconversion material-labeled sample is excited and transmitted through the multi-channel filtering module to the tube lens to form an image. The microlens array is located at the focal point of the tube lens, and the sensor is located at the focal length of the microlens array for recording the three-color imaging of the sample.

The present invention discloses a multi-color synchronous reconstruction method for a light field microscopy system, belonging to the fields of computer vision and digital image processing. The method includes: A1: extracting spectral information from multi-wavelength fluorescence light field images and performing spectral correction according to the wavelength response function of the image acquisition system; A2: separating single-wavelength light field images and performing reconstruction one by one using the single-wavelength light field microscopy reconstruction algorithm; A3: coloring the single-wavelength reconstruction results and fusing all the reconstruction results according to the principle of linear superposition. The present invention interprets the synchronous multi-color light field reconstruction method as three sub-methods: spectral information extraction and correction, monochromatic reconstruction, and image coloring and fusion, and proposes a reliable multi-color reconstruction method that is easy to implement and expand, which is of great significance for promoting the research of neuroscience and the development of synchronous multi-color light field microscopy technology.

The present invention discloses a method, device and medium for calibrating the step size of a two-dimensional scanning module in a scanning light field imaging system, which relates to the technical field of light field microscopy imaging. The method includes: collecting two-dimensional original scanning light field data with an initial scanning step size and rearranging it into four-dimensional phase space data to obtain a multi-view rearranged image; selecting the middle view of all views in the multi-view rearranged image to obtain a non-scanning center view sub-image; calculating a sub-pixel displacement matrix of all non-scanning center view sub-images relative to the first non-scanning center view sub-image; calculating an average displacement amount according to the sub-pixel displacement matrix, and calculating a step size adjustment coefficient according to the average displacement amount; updating the correct scanning step size according to the step size adjustment coefficient. The present invention calibrates the scanning step size of two-dimensional scanning imaging with higher speed and higher precision, and improves the spatial resolution of scanning imaging.

A method for automatically determining whether to perform denoising includes the following steps: S1: scanning a sample using a light field microscopy system to obtain a light field microscopic image containing 4-dimensional information, where the sample is a sample to be 3D reconstructed; S2: rearranging the light field microscopic image to obtain a multi-view rearranged image; S3: calculating a cleanliness score of the multi-view rearranged image; and determining whether to perform denoising process on the multi-view rearranged image, based on the cleanliness score of the multi-view rearranged image.

The present invention discloses a method for automatically judging whether to perform denoising, comprising the following steps: S1: scanning a sample using a light field microscopy system to obtain a light field microscopy image with 4-dimensional information; wherein the sample is a sample to be 3D reconstructed; S2: rearranging the light field microscopy image to obtain a multi-view rearranged image; S3: calculating a cleanliness score of the multi-view rearranged image; judging whether to perform denoising on the multi-view rearranged image based on the cleanliness score of the multi-view rearranged image. The present invention can automatically judge whether to perform denoising, which is not only efficient but also more objective in judgment results compared to manual judgment.