415 results about "Fluorescence microscope" patented technology

A deep learning-based digital / virtual staining method and system enables the creation of digitally / virtually-stained microscopic images from label or stain-free samples. In one embodiment, the method of generates digitally / virtually-stained microscope images of label-free or unstained samples using fluorescence lifetime (FLIM) image(s) of the sample(s) using a fluorescence microscope. In another embodiment, a digital / virtual autofocusing method is provided that uses machine learning to generate a microscope image with improved focus using a trained, deep neural network. In another embodiment, a trained deep neural network generates digitally / virtually stained microscopic images of a label-free or unstained sample obtained with a microscope having multiple different stains. The multiple stains in the output image or sub-regions thereof are substantially equivalent to the corresponding microscopic images or image sub-regions of the same sample that has been histochemically stained.

The present invention relates to an image acquisition device and acquisition method for detecting a target material signal, which have improved sensitivity and uniformity compared to those of a conventional fluorescence microscope used for acquiring fluorescence images. The present invention provides the image acquisition device for detecting a target material signal, comprising: (a) a camera for detecting a fluorescence signal; (b) light sources that are disposed outside the camera at regular intervals in a circular shape; (c) a shielding guide that is positioned above the light sources and which transmit only vertical direction light of light emitted from the light sources; and (d) a prism that refracts the light passing through the shielding guide toward a sample.

The invention belongs to the field of biological pharmacy, and relates to a temperature-sensitive hydrogel, in particular to a temperature-sensitive hydrogel SV (at) HP based on hyaluronic acid as well as a preparation method and application of the temperature-sensitive hydrogel SV (at) HP. The temperature-sensitive hydrogel SV (at) HP entraps CS-siRNA nanoparticles and VEGF (at) PLGA microspheres, and is used for realizing microenvironment responsive drug delivery to treat intrauterine adhesion. Chlorogenic acid and TGF beta1 siRNA are self-assembled through disulfide bonds to form CS-siRNA nano-particles, the CS-siRNA nano-particles are used for treating intrauterine adhesion through gene and chemical therapy, and cell and animal experiments prove that the CS-siRNA nano-particles have a good effect of improving intrauterine adhesion. Through evaluation of a real-time fluorescence quantification method, a western blot method, a cell scratch experiment, laser confocal microscope observation, inverted fluorescence microscope observation, an in-vivo experiment and the like, the SV-coated HP gel has the effects of resisting inflammation, improving endometrial fibrosis, promoting IUA rat endometrial angiogenesis and repair, inhibiting intrauterine adhesion and improving the intrauterine environment.

The invention discloses an automatic focusing method, device and equipment for a fluorescence microscope. The method comprises the following steps: S10, acquiring an image acquired by the microscope and extracting a target cell contour in the image acquired by the microscope; s20, the contour of the target cell is identified, an internal area and an external area are obtained, the internal area is a cell main body part, and the external area is a part except the cell main body part; s30, the definition value of the internal area and the definition value of the external area are calculated respectively, weighting processing is carried out respectively, and the weight corresponding to the definition value of the internal area is larger than the weight corresponding to the definition value of the external area; s40, calculating a global definition score according to a preset formula; and S50, automatically adjusting the focal length of the microscope based on the global definition score. The definition values of the internal region and the external region are respectively calculated and weighted, and the detail definition of the cell main body part is preferentially considered, so that the interference of the halo effect on the focusing result is effectively reduced, and the focusing precision and the imaging quality are improved.

The invention discloses a biological experiment automatic touch-free adding system based on optical control and an operation method thereof, and relates to the technical field of biological experiment equipment, the system comprises the following components: an optical marking module, an optical imaging and detection module, an optical control module and an automatic control module; high automation of biological experiments is realized by integrating a plurality of modules including optical marking, optical imaging and detection, optical control and automatic control, efficient and stable marking of biological samples is ensured by utilizing specific marking of a quantum dot solution and combining accurate control of a microfluidic channel in the optical marking stage, and then, the accuracy of the biological samples is ensured. Through advanced equipment such as a high-resolution fluorescence microscope and an optical spectrum analyzer, the system can accurately obtain optical information of a marked biological sample, a solid foundation is provided for subsequent control and recognition, in the optical control stage, the optical tweezers system achieves accurate control over the biological sample in a touch-free mode, and the accuracy of the biological sample is improved. Pollution and damage possibly caused by traditional physical contact are avoided.

The invention discloses a method and a device for detecting errors of a rotary table and a rotary shaft of an overall measurement light path connected upper rotary shaft type laser tracker, and belongs to the technical field of laser tracking measurement. The device comprises a microscope, a fluorescent microsphere, a substrate and a six-dimensional precision calibration displacement table, the microscope is located above the fluorescent microsphere, and the six-dimensional precision calibration displacement table is located above the substrate. The fluorescent microspheres are fixed on the substrate, and the substrate is fixed on the six-dimensional precise calibration displacement table. By combining the fluorescence microscope with the astigmatism imaging function, the precise displacement table, space coordinate calculation and other operations, the determination of the axis space linear equation of the upper and lower rotary tables of the whole measurement light path connected upper rotary shaft type laser tracker and the detection of the shaft radial runout in a complete rotation period are realized under one coordinate system; a necessary technical foundation is laid for realizing error compensation of the rotary table and the rotary shaft of the rotary shaft type laser tracker connected with an overall measurement light path and improving the tracking precision and the stability of the laser tracker.

The invention discloses an oral risk assessment method and system based on image recognition and a medium, and relates to the technical field of image recognition, and the method comprises the steps: carrying out the block segmentation of a user oral image, activating a lesion recognizer to recognize the features of an image block, carrying out the matching of an oral disease, and obtaining a corresponding flora type set; imaging the target flora by using a fluorescence microscope, and extracting fluorescence characteristics; and generating an oral risk decision of the target user in combination with the identified disease information and the fluorescence characteristics. Therefore, the technical effects of multi-dimensional image comprehensive analysis and evaluation accuracy and efficiency improvement are achieved.

A fluorescence microscopy inspection system includes light sources able to emit light that causes a specimen to fluoresce and light that does not cause a specimen to fluoresce. The emitted light is directed through one or more filters and objective channels towards a specimen. A ring of lights projects light at the specimen at an oblique angle through a darkfield channel. One of the filters may modify the light to match a predetermined bandgap energy associated with the specimen and another filter may filter wavelengths of light reflected from the specimen and to a camera. The camera may produce an image from the received light and specimen classification and feature analysis may be performed on the image.

The present application discloses methods for characterising vesicles. The method involves (1) a sample preparation step, comprising providing a test specimen with vesicles attached to a substrate, wherein the vesicles are labelled with one or more fluorescent probes; (2) an image acquisition step, comprising imaging said one or more fluorescent probes on the vesicles to generate image data; (3) an image processing step which identifies individual vesicles and constructs a feature vector containing characterising parameters for individual vesicles characterising parameters (including a morphological parameter) (4) a data transformation step to calculate modified feature vectors of lower dimensionality for individual vesicles; and (5) a characterisation step, which characterises the vesicles based on the modified feature vectors. The application also discloses methods for immobilising vesicles on a substrate, as well as substrates functionalised to capture vesicles.

The invention discloses an ANA detection image-text report intelligent generation method and system, and relates to the technical field of immunodetection, the system uses different fluorescent markers to mark ANA, signal sensitivity is enhanced, and detection accuracy is ensured. Then, collecting fluorescence signals of different wavelengths by adopting a fluorescence microscope and a multi-wavelength excitation light source, and obtaining dynamic change data of reaction intensity by combining a time-resolved fluorescence imaging technology to generate a multi-channel imaging data set; thirdly, noise removal, signal enhancement and image alignment are carried out through an image preprocessing technology, and data consistency is ensured; based on a multi-channel data set, a convolutional neural network is used for training and testing an initial model, an intelligent detection model is constructed, a fluorescence reaction mode is extracted, finally, a detailed ANA detection report is automatically generated by calculating a comprehensive scoring coefficient and comparing the comprehensive scoring coefficient with a preset threshold value, and an accurate immunodiagnosis basis is provided for doctors.

The invention discloses a microscopic image integer pixel alignment method based on sub-pixel edge offset detection, and relates to the field of microscopic image processing. The problems that an existing sub-pixel registration technology depends on iterative optimization or a complex deformation model, and real-time processing is difficult; sensor sampling errors and mechanical translation errors need to be compensated through a high-resolution camera or a precise displacement table, and the system cost is high. According to the invention, through sub-pixel-level offset compensation, sub-pixel alignment can be realized without depending on a high-precision displacement table, and the hardware cost is remarkably reduced. The method comprises the following steps: extracting image edge feature points by using an edge detection algorithm, fitting a gray centroid to calculate a sub-pixel offset, and counting an average offset; generating an affine transformation matrix according to the average offset, resampling the image, and aligning pixel values to an integer coordinate grid; and finally, cutting the image edge according to the interpolation algorithm type to eliminate the ghost. The method is also suitable for the application fields of fluorescence microscope multi-view splicing, super-resolution reconstruction and the like.

A deep learning-based digital / virtual staining method and system enables the creation of digitally / virtually-stained microscopic images from label or stain-free samples. In one embodiment, the method of generates digitally / virtually-stained microscope images of label-free or unstained samples using fluorescence lifetime (FLIM) image(s) of the sample(s) using a fluorescence microscope. In another embodiment, a digital / virtual autofocusing method is provided that uses machine learning to generate a microscope image with improved focus using a trained, deep neural network. In another embodiment, a trained deep neural network generates digitally / virtually stained microscopic images of a label-free or unstained sample obtained with a microscope having multiple different stains. The multiple stains in the output image or sub-regions thereof are substantially equivalent to the corresponding microscopic images or image sub-regions of the same sample that has been histochemically stained.

The present invention discloses a method and system for fusing visible light images and fluorescence images, and relates to the field of medical image processing technology. The method comprises: performing image registration on a visible light camera and a near-infrared camera to obtain registration parameters; synchronously acquiring visible light images and fluorescence images, preprocessing the received fluorescence images, and aligning the pixels of the visible light images and the preprocessed fluorescence images according to the registration parameters; performing pixel fusion on the aligned visible light images and fluorescence images to generate a multispectral image; and sequentially merging the fused multispectral images to generate a corresponding video sequence. The present invention adopts offline camera registration, and utilizes the registration parameters to efficiently and accurately align visible light images and fluorescence images. By enhancing the preprocessing, a high-quality fluorescence image can be generated. The fusion weight is calculated based on the edge intensity value and the fluorescence intensity value, so that the weighted fused multispectral image can simultaneously maintain edge and fluorescence intensity information, and is suitable for medical equipment such as fluorescence microscopes / endoscopes.

A method for automatically ascertaining illumination brightnesses to be adjusted of at least two light sources for exciting at least one respective fluorophore in a sample to be imaged in a fluorescence microscope includes separately controlling, in terms of illumination brightness, each of the at least two light sources, detecting an image intensity of a microscopically imaged sample with at least two detectors, and automatically ascertaining the illumination brightnesses to be adjusted of the at least two light sources in such a way that a predefined setpoint of a signal-to-noise ratio is reached for each fluorophore. In order to ascertain the illumination brightnesses of the at least two light sources, cross-talk of a detector for different emission spectra of the fluorophores and / or cross-excitation of a fluorophore for different illumination spectra of the light sources are / is taken into account.

The invention discloses a denoising method based on R2U-Net and an adaptive noise addition model, and the method comprises the steps: obtaining an original noise image from a fluorescence microscope, carrying out the preprocessing, and reducing the noise level of the original noise image through median filtering; building a noise adding module to add noise to the image, wherein the noise adding comprises regional noise adding and global noise adding; the method comprises the following steps: establishing an R2U-Net network, and introducing a residual learning and recursion unit on the basis of U-Net; the R2U-Net network is trained, a new noise image is generated through self-supervised learning and serves as input of the R2U-Net network, the R2U-Net network is used for predicting a denoising result and optimizing the network, and the optimized R2U-Net network is used for denoising an image to be processed. According to the method, the noise in the fluorescent microscopic image can be dynamically and accurately removed without a large amount of annotation data, the detail information of the image is reserved, and the image quality is improved.

This disclosure provides microscope-based systems and methods that significantly improve image contrast and have better vertical / horizontal resolution. This disclosure uses confocal microscopy in replacement of conventional fluorescent microscopy, for example, adding a spinning disk or a confocal microscopy scanning unit on the prior invention. Using a spinning disk vastly improves the speed of image acquisition (allowing for imaging of fast dynamic processes and live specimens), and considerably reduces photo damage. Disclosed herein are some embodiments to demonstrate how to set up the spinning disk microscopy system for image-guided illumination and photolabeling.

The invention discloses a fixing reagent and method for double-color fluorescence detection of an anti-NMDAR antibody and application, the fixing reagent comprises paraformaldehyde and a mixture of methanol and acetone, and the mixture comprises 10-25% by volume of methanol, 75-90% by volume of acetone, 10-25% by volume of paraformaldehyde, 10-25% by volume of paraformaldehyde and 10-25% by volume of paraformaldehyde. The fixing effect is better by adopting paraformaldehyde with the mass-volume ratio concentration of 4% and a mixture of methanol with the volume fraction of 15% and acetone with the volume fraction of 85% in two steps. By adopting the fixing liquid and the fixing method, the antigenicity of NMDAR cannot be damaged, obvious green fluorescence cells can be seen in a positive sample, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed by using a fluorescence microscope.

The invention belongs to the field of molecular sieve material characterization, and particularly relates to a method for analyzing aluminum atom pairs in a ZSM-5 molecular sieve based on super-resolution fluorescence microscopic imaging, which comprises the following steps: activating a ZSM-5 molecular sieve sample; contacting the obtained sample with probe molecules; washing the ZSM-5 molecular sieve sample, and removing unbound or nonspecifically bound probe molecules; imaging the marked sample by using a super-resolution fluorescence microscope to obtain an image sequence containing a single fluorescent molecule emission event; processing the image sequence, determining a space coordinate of each fluorescence event, and reconstructing a super-resolution fluorescence image exceeding an optical diffraction limit; and based on the space coordinates of the fluorescent dots in the super-resolution fluorescence image, carrying out statistics to obtain the position, surface density or distribution information of the aluminum atom pairs in the ZSM-5 molecular sieve. According to the method disclosed by the invention, nanoscale spatial resolution, direct imaging and quantitative analysis of Al-pair in ZSM-5 (Zeolite Socony Mobil-5) can be realized.

2.1. To increase the resolution of quasi-confocal line scanning microscopes with a main beam splitter for coupling the illumination and detection beam paths, the "re-scan" method involves descanning the sample light in the detection beam path by the illuminating beam deflection unit. After descanning, the sample light is scanned again in the detection beam path by moving it across the sensor behind the confocal aperture by a second beam deflection unit in a direction transverse to the linear focus. To increase resolution in the longitudinal direction, the illumination line can be structured. This optical arrangement has the disadvantage of significant light losses due to the large number of optical interfaces that must be passed through. 2.2. A microscope in which the illuminating beam deflection unit is optically positioned between the light source and the main beam splitter, allowing the sample light to reach the sensor away from the illuminating beam deflection unit (non-descanned), is more light-sensitive and flexible. A control unit, in one operating mode, moves the sample light beam across the sensor using only the second beam deflection unit. Depending on the operating mode, synchronization with the first beam deflection unit may be provided. 2.3. Due to its high light sensitivity, the microscope is particularly suitable for fluorescence microscopy.

The invention discloses an image recognition analysis method and system for heart macrophage inflammation regulation and control, and relates to the technical field of macrophage inflammation regulation and control recognition, and the method comprises the following steps: observing and obtaining a cell image through a fluorescence microscope, and preprocessing the cell image to obtain a preprocessed image; constructing a cell recognition deep learning model, and training the cell recognition deep learning model by extracting fluorescence features of macrophages and gray features of inflammatory cells; carrying out statistics on macrophages and inflammatory cells in the inflammation area, and outputting a regulation and control inspection result; the method is used for solving the problem that the judgment of doctors on the condition of cardiac inflammation is easily affected due to insufficient recognition accuracy of macrophages and inflammatory cells in the existing macrophage inflammation regulation and control recognition technology.

The invention discloses a method for characterizing the degree of fusion of new and old asphalt based on fluorescence microscopy, and the process is as follows: using a confocal microscope to observe the surface of a wood simulated aggregate coated with homogenous SBS modified asphalt, obtaining fluorescence images at different depths, calculating an average fluorescence intensity curve, and fitting a fluorescence intensity attenuation correction function; observing the mixed asphalt on the surface of the wood simulated aggregate under the confocal microscope; scanning from the outermost layer to the innermost layer of the wood simulated aggregate, and outputting a plurality of fluorescence images of the new and old asphalt in the asphalt film thickness layer; performing noise reduction processing on the fluorescence image, obtaining a concentration change curve of the new asphalt and correcting it using an attenuation correction function; calculating a concentration attenuation change curve of the new asphalt, judging the influence of noise and the asphalt fusion layer, retaining the layer position with a fitting coefficient less than or equal to 95%, and calculating the concentration attenuation change curve after removing the influence of noise and fluorescence signal attenuation; and defining the degree of fusion of the mixed asphalt by an area method.

A method of analyzing a mixed fluorescence response of a plurality of fluorophores in a microscopic sample includes reconstructing individual fluorescence responses from a mixed fluorescence response using spectral un-mixing based on reference emission spectra for fluorophores to be reconstructed, and a procedure for determining and validating reference emission spectra including providing a plurality of image acquisition settings for a sequence of images of the sample equal to, or greater than, the plurality of fluorophores and including an illumination setting for each image, acquiring the sequence of images using the plurality of image acquisition settings and storing each image together with the corresponding illumination setting, determining candidate reference emission spectra for the fluorophores to be reconstructed from the sequence of images of the sample using one or more reference emission spectra determination algorithms, and conditionally using the candidate reference emission spectra as the reference emission spectra in the spectral un-mixing.

The invention discloses a detection method of mycobacteria. The detection method comprises the following steps: S1, constructing a fusion protein gp23RD-GFP; s2, a target protein is obtained from the fusion protein gp23RD-GFP; s3, mixing the target protein with the mycobacterium culture, and incubating; s4, preparing a microscopic examination sample; s5, carrying out microscopic examination by using a fluorescence microscope; according to the method, a functional truncated body of phage TM4 receptor binding protein gp23 is fused with a green fluorescent protein to construct a fusion protein gp23RD-GFP, the fusion protein gp23RD-GFP is purified to obtain a target protein, the target protein and a mycobacterium culture are mixed and incubated, and finally the incubated mycobacterium culture is subjected to microscopic examination sample preparation. The detection mode is simple and rapid, and support can be provided for the joint development of the subsequent mycobacterium detection technology.

Devices, systems, and methods are provided for in vivo fluorescence imaging. Disclosed herein is an implantable miniature fluorescence imager on a chip having a custom imaging array with angle selective gratings, fiber optics, or microcollimators for image deblurring, and optical filters that can be tuned to image fluorescence from multiple fluorophores simultaneously. Power is supplied by an on-chip power source or transmitted to the chip from an external transducer such as an ultrasound transducer, electromagnetic transducer, inductive transducer, or radiofrequency transducer. Wireless communication may be provided by electromagnetic or ultrasound links to the device. The function of a fluorescence microscope is provided in a millimeter-scale device that can be readily implanted in tissue and used to image fluorescently labeled cells in vivo. The small size of the fluorescence imager makes possible sustained in vivo imaging with real-time monitoring of multiple cell types within Shifting the dynamic diseased tissue or a tumor.

A system and method for analyzing mechanical properties of biological tissues combines light-sheet fluorescence microscopy with mechanical perturbation capabilities. The system integrates a MEMS-based compression device, high-speed light-sheet scanner, and digital image correlation analysis to characterize viscoelastic properties of three-dimensional cellular structures. The system enables quantification of spatially-resolved mechanical characteristics and cellular responses through volumetric strain analysis, correlating mechanical properties with fluorescence-based cellular markers. Implementation with maternal-fetal interface models demonstrates capabilities in analyzing tissue mechanics during cellular invasion processes. The system provides applications in studying mechanical interactions in biological processes including embryonic development, cancer metastasis, and wound healing. The disclosed methodology enables unprecedented analysis of mechanical property dynamics at cellular interfaces in three-dimensional tissue environments.

The application belongs to the technical field of biology and specifically relates to a method for analyzing glycans on single extracellular vesicles in a fluid sample. The method uses magnetic nanoparticles to capture and manipulate EVs, based on the specific affinity of lectins for specific glycan structures, uses polydisperse microdroplets to separate the captured EVs, realizes local signal amplification in the droplets through enzymatic reaction, adopts fluorescence microscopy to collect images of the droplets, and thus realizes identification and analysis of the glycan components on single EVs. The preparation process of the microdroplets formed by the oscillating emulsification method of the application makes the entire detection process more simple and fast, does not require any microfluidic chip and special device, and can be completed in a centrifugal tube, and all the microdroplets are generated in parallel and quickly, and the amount of microdroplets can be flexibly expanded. Moreover, compared with the traditional ELISA, the method has high sensitivity, low detection limit and small sample consumption.

The invention discloses an online optical clear solution for three-dimensional structure imaging of a large-volume biological sample, which is characterized in that the solution comprises iohexol, urea, 1, 2-propylene glycol, dimethyl sulfoxide, 3-[3-(cholamidopropyl) dimethylamino] propanesulfonic acid inner salt and water. The invention further discloses a three-dimensional imaging method for the large-volume biological sample. The method comprises the following steps: carrying out transparentizing treatment on the biological sample by using the online optical transparentizing solution, and carrying out three-dimensional imaging on the transparentized biological sample by using a fluorescence microscope. The fluorescence microscope comprises a confocal microscope and / or a light sheet microscope. When the online optical transparent solution is used for imaging, the transparent depth of a surface sample is large, the single imaging depth can be increased, and the cutting thickness can be improved, so that the cutting frequency is reduced, and finally the data acquisition efficiency of a large-volume sample is improved.

The invention discloses a ctEVs separation and analysis method with time resolution capability and application thereof. The ctEVs separation and analysis method comprises the following steps: S1, culturing cells by adopting glycometabolism markers; or injecting the animal by adopting a glycometabolism marker; step S2, manufacturing a fishbone-shaped micro-fluidic chip modified by streptavidin; s3, separating and analyzing the new tumor source extracellular vesicles by the micro-fluidic chip, and co-incubating the extracellular vesicles and the aptamer probe co-modified by Biotin and DBCO to realize biotinylation of the new tumor source extracellular vesicles; the aptamer probe is a heterologous multivalent aptamer probe; step S4, capturing biotin-labeled target extracellular vesicles by adopting a chip; and S5, acquiring a chip image by using a fluorescence microscope, and analyzing the concentration of the target extracellular vesicles in the sample according to a fluorescence signal. The method can be used for screening tumor circadian rhythm markers and the like.

The invention discloses a serum migration body identification method which comprises the following steps: extracting a serum migration body from serum by performing multiple centrifugal treatment on the serum; carrying out immunofluorescence staining treatment on the serum migration body to obtain an immunofluorescence stained serum migration body; and carrying out imaging treatment on the serum migration body subjected to immunofluorescence staining by utilizing a super-resolution fluorescence microscope SIM to identify the serum migration body. Whether an extracellular vesicle in serum is a migration body or not is identified from the two aspects of biochemistry and shape, so that the defect that the serum migration body is identified only from the shape in the prior art is overcome.

The invention discloses an electrochemical microscopic imaging analysis method based on a fluorescence signal, which comprises the following steps of: constructing a fluorescence response electrochemical signal instrument by combining electrochemistry with a fluorescence microscope, and observing the redox potential of K3 [Fe (CN) 6] by taking a terpyridyl ruthenium (Ru (bpy) 32 +) / potassium ferricyanide (K3 [Fe (CN) 6]) fluorescence system as a research object. Influences of electrochemical behaviors on fluorescence signals are deeply explored, and the relationship between fluorescence signal changes and electric signal changes is revealed. According to the present invention, the combination of the electrochemical technology and the fluorescence signal is achieved, the disadvantage of the poor spatial-temporal resolution of the electrochemiluminescence is made up, the new dimension capable of being regulated and controlled by the electrical parameter is provided for the fluorescence imaging, and the Faraday current and the non-Faraday current can be completely reflected by the fluorescence signal. Therefore, the method has wide application value in the aspects of exploring the conductivity of the nano material and the like.