1272 results about "Immunofluorescence" patented technology

Clinical information, molecular information and/or computer-generated morphometric information is used in a predictive model for predicting the occurrence of a medical condition. In an embodiment, a model predicts whether a patient is likely to have a favorable pathological stage of prostate cancer, where the model is based on features including one or more (e.g., all) of preoperative PSA, Gleason Score, a measurement of expression of androgen receptor (AR) in epithelial and stromal nuclei and/or a measurement of expression of Ki67-positive epithelial nuclei, a morphometric measurement of a ratio of area of epithelial nuclei outside gland units to area of epithelial nuclei within gland units, and a morphometric measurement of area of epithelial nuclei distributed away from gland units. In some embodiments, quantitative measurements of protein expression in cell lines are utilized to objectively assess assay (e.g., multiplex immunofluorescence (IF)) performance and/or to normalize features for use within a predictive model.

The present invention provides methods and compositions for isolating and detecting rare cells from a biological sample containing other types of cells. In particular, the present invention includes a debulking step that uses a microfabricated filters for filtering fluid samples and the enriched rare cells can be used in a downstream process such as identifies, characterizes or even grown in culture or used in other ways. The invention also include a method of determining the aggressiveness of the tumor or of the number or proportion of cancer cells in the enriched sample by detecting the presence or amount of telomerase activity or telomerase nucleic acid or telomerase expression after enrichment of rare cells. This invention further provides an efficient and rapid method to specifically remove red blood cells as well as white blood cells from a biological sample containing at least one of each of red blood cells and white blood cells, resulting in the enrichment of rare target cells including circulating tumor cells (CTC), stromal cells, mesenchymal cells, endothelial cells, fetal cells, stem cells, non-hematopoietic cells etc from a blood sample. The method is based upon combination of immuno-microparticles (antibody coated microparticles) and density-based separation. The final enriched target cells can be subjected to a variety of analysis and manipulations, such as flowcytometry, PCR, immunofluorescence, immunocytochemistry, image analysis, enzymatic assays, gene expression profiling analysis, efficacy tests of therapeutics, culturing of enriched rare cells, and therapeutic use of enriched rare cells. In addition, depleted plasma protein and white blood cells can be optionally recovered, and subjected to other analysis such as inflammation studies, gene expression profiling, etc.

Apparatus, methods, and computer-readable media are provided for segmentation, processing (e.g., preprocessing and/or postprocessing), and/or feature extraction from tissue images such as, for example, images of nuclei and/or cytoplasm. Tissue images processed by various embodiments described herein may be generated by Hematoxylin and Eosin (H&E) staining, immunofluorescence (IF) detection, immunohistochemistry (IHC), similar and/or related staining processes, and/or other processes. Predictive features described herein may be provided for use in, for example, one or more predictive models for treating, diagnosing, and/or predicting the occurrence (e.g., recurrence) of one or more medical conditions such as, for example, cancer or other types of disease.

The invention discloses a method and an automatic instrument device for enriching and extracting target cells and separating single cells by using a positive magnetic bead method and performing immunity and molecular biology identification and analysis on single cells. By the method and the instrument device, the on/off of a capture magnet and a release magnet is controlled by various methods to complete target cell searching, capturing, cleaning and releasing operation once or for multiple times, so that the target cell detection sensitivity and stability are improved. When the capture magnet searches and captures the target cells, the search line is circular, square, comb-shaped, S-shaped or U-shaped. In addition, the captured substances are filtered, most free micro magnetic beads are removed, the purity of the product is further improved and the product can be used as a good experimental material. By combining a special filter and the adsorption of the capture magnet, more than 95 percent of free micro magnetic beads can be effectively removed. Meanwhile, the types of the single cells are identified and biological characteristics of the single cells are analyzed by an immunofluorescence staining method and a method in molecular biology, and effective biological indexes are provided for clinical diagnosis and treatment of cancers.

The invention discloses a magnetic fluorescent kit for rapidly detecting microbes as well as a preparation method and a use method thereof. The kit comprises two components: 1) immunomagnetic microspheres specifically bound with the microbes to be detected; and 2) immunofluorescent microspheres specifically bound with the microbes to be detected. The preparation method comprises the following steps: (1) preparation of the immunomagnetic microspheres; and (2) preparation of the immunofluorescent microspheres. The use method comprises the following steps: (1) adding the sample to be detected, lyophilized powder of the immunomagnetic microspheres and the immunofluorescent microspheres to a buffer solution; (2) ensuring the surfaces of the identified microbes to have antigenic determinants simultaneously bound with the immunomagnetic microspheres and the immunofluorescent microspheres; (3) enriching the microbes bound with the immunomagnetic microspheres through magnetic separation of the immunomagnetic microspheres; and (4) qualitatively and quantitatively judging the microbes by measuring the fluorescence intensity of the immunofluorescent microspheres bound with the separated and enriched microbes. The kit, the preparation method and the use method have the advantages of rapidness, quantitative property and wide scope of application.

The invention discloses immunofluorescence chromatography test paper for CRP (C-reaction protein)/SAA (Serum amyloid A protein) quantitative combined detection, aiming at providing a test strip capable of realizing quantitative combined detection on the content of CRP/SAA in human peripheral blood and quantitative combined detection on the content of CRP/SAA in venous blood. The immunofluorescence chromatography test paper is technically characterized by comprising a box with a cover, wherein a detection test strip and a blood diluent bottle are arranged in the box, the detection test strip comprises a bottom lining, the bottom lining is provided with a nitrocellulose membrane, one end of the nitrocellulose membrane is connected with a fluorescent microsphere marked antibody fixation pad, the fluorescent microsphere marked antibody fixation pad is connected with a sample pad, the other end of the nitrocellulose membrane is connected with an absorption pad, the fluorescent microsphere marked antibody fixation pad is coated by a CRP monoclonal antibody and an SAA monoclonal antibody, a CRP detection line coated by a CRP monoclonal antibody, an SAA detection line coated by an SAA monoclonal antibody, and a quality control line coated by a goat-anti-mouse IgG polyclonal antibody are arranged on the nitrocellulose membrane in parallel. The immunofluorescence chromatography test paper belongs to the technical field of biological medicines.

The invention relates to a quantum-dot-labeled immunofluorescence multi-antigen simultaneous detection method, belonging to the detection technical field. The method of the invention comprises the steps that the objects, i.e., the antibodies of the antigen to be detected are respectively connected to the quantum dot and the nano-particle like magnetic nano-particle by using the fluorescence characteristics of the quantum dot, like multiple-wavelength excitation, high-strength fluorescence emission, narrow emission peak, symmetrical peak shape, and stable luminescence; the antigen to be detected and an excessive amount of quantum-dot-labeled antibody are added; by immune reaction, the objects , i.e., the antibodies of the antigen to be detected, are jointed with the antibodies fixed on the quantum dot and the nano-particle like magnetic nano-particle to result in the joint of the quantum dot and the nano-particle like magnetic nano-particle, so as to form the composition of the antigen and the antibody, i.e., complex; the complex and the unreacted quantum-dot-labeled substance, i.e., free substance are separated, the intensity of fluorescence signal of the free substance is detected, and quantitative detection is carried out for the antigen to be detected. The invention has high sensitivity and wide detection range, and can simultaneously detect multiple antigens with simple operation and at a low cost.

The invention discloses a micro-fluidic chip immunofluerescence rapid detection kit as well as preparation and detection methods thereof. The kit comprises a micro-fluidic chip; the micro-fluidic chip comprises one or more detection units; the detection units are arranged on a radius axis of the micro-fluidic chip and are arranged in a divergence manner by taking the micro-fluidic chip as a center point; and a sample feeding groove, a fluorescence probe groove, a reaction detection groove, a waste liquid groove and microchannels which are mutually connected are sequentially arranged on the detection units from the center point of the chip to the edge. By using the kit, the dosage of a reagent is small, the integration degree is high, and a reaction systems is a homogeneous phase; and all the reagents are solidified in the chip in advance, and antigen-antibody reaction is carried out in a relatively closed space, so that crossed contamination and environmental pollution do not occur, and the kit has the advantages of high efficiency, high flux, high sensitivity, rapidness, sensitiveness, stability and the like.

This invention presents improved methodology for identification of nucleated cells in flow cytometric analysis when immunofluorescent dyes are also used. Briefly, in the method nucleic acids are stained with a fluorescent dye which can then be used to identify the nucleated cells by measurement of fluorescence on a flow cytometer. The improvement presented by this invention is the use of a saturating (or near saturating) amount of a nucleic acid dye, or mixture of dyes, which gives low fluorescence at excitation conditions, so as not to greatly interfere with the signals of the immunofluorescent dyes.

The invention relates to a method for screening an aptamer specifically bound with an alpha-fetoprotein by using a capillary electrophoresis technology. The method comprises the following steps of designing and establishing a random oligonucleotide library; enriching oligonucleotides which can be bound with alpha-fetoprotein by combining a capillary electrophoresis technology and an SELEX (Systematic Evolution of Ligands by Exponential Enrichment) screening method; and cloning, sequencing, and carrying out capillary electrophoresis analysis and immunofluorescence verification to finally obtain oligonucleotides which can be specifically bound with the alpha-fetoprotein, namely the aptamer. The invention provides an efficient and specific aptamer screening method. The method has the advantages that the screening time is shortened by means of the advantage of the capillary electrophoresis technology, and the screening operation can be finished through four rounds of circulation; and meanwhile, the non-specific screening caused by a matrix effect is reduced.

The invention discloses a new method for detecting non-humoral rare karyotes. Non-humoral rare karyotes in a biological body fluid are obtained through many means, and the combined use of the antibody of a humoral cell surface marker and the specific probe of the non-humoral rare karyotes is carried out to discriminate the enriched rare karyotes. Compared with commercial discriminating methods existing in the present market, the method disclosed in the invention well combines a fluorescent in situ hybridization (FISH) method for processing the non-humoral rare karyotes with the immunofluorescence dyeing for processing humoral rare karyotes, realizes the systemic associative processing of a same specimen sheet, and can realize the intelligible observation of two processing results.

The invention discloses a sealing and stabilizing agent for a microporous board. In terms of 100ml water, 0.1-3g of protein, 3-20g of saccharide, 0.01-1g of organic polymer, 0.01-0.05ml of nonionic surfactant, 0.01-0.1g of preservative and 10-100mmol of buffer solution are contained. The sealing and stabilizing agent disclosed by the invention is researched and designed on account of the characteristic that antigen or antibodies coated on a stationary phase are easy to inactivate. The sealing and stabilizing agent includes not only sealing components but also organic components and inorganic components, wherein the sealing components can be used for effectively sealing excessive loci on a sealing board, and the organic components and the inorganic component can be used for effectively stabilizing the antigen or the antibodies, so that the sealing purpose and the stabilizing purpose can be organically combined together through one-step operation. The sealing and stabilizing agent disclosed by the invention can be applied to immunological detection methods such as enzymelinked immunosorbent assay, chemiluminesent immunoassay, time resolved fluoroimmunoassay and the like, wherein the microporous board serves as the stationary phase; and the sealing and stabilizing agent can be used for effectively stabilizing the activity of the antigen or the antibodies coated on a stabilizing board.

A method of preparing an antigen-immobilized immuno-fluorescence slide, the method comprising: immobilizing a C-reactive protein on a slide to prepare a protein chip; mixing an antibody that specifically binds to a target protein, with streptavidin to label the antibody with a fluorescent nanoparticle; immuno-reacting the antibody by competitive mixing, assaying with a fluorescence camera, wherein the immobilizing of the C-reactive protein on the slide comprises: modifying the slide with 3-aminopropyltrimethoxysilane to prepare a modified slide; hydrating the slide modified with 3-aminopropyltrimethoxysilane; activating the modified slide by using a glutaraldehyde solution; dissolving a C-reactive protein at a concentration of 0.01-0.5 mg/ml in a 30-70 mM phosphate buffer solution (pH 6.5-7.8) to prepare an antigen solution for immobilization; placing a petri dish comprising the slide on a spotting guide and spotting 1-100 μl of the antigen solution on spotting points; and performing a reaction on the slide prepared as described above for 1-6 hours to immobilize the antigen, and an immune-fluorescence slide prepared by using the method.

The invention relates to a cell chemical dyeing method compatible with immunofluorescence analysis and relative applications. The invention further relates to a method for checking epithelial tissue source tumor cell in human peripheral blood sample, which uses the cell chemical dyeing method compatiable with immunofluorescence analysis. The inventive check method can simply, quickly and accurately check the tumor cells of epithelial tissue source in blood circulation from the karyocyte separated from small human periphery blood sample.

The invention discloses an immunofluorescence detection test strip and a preparation method thereof for rapid quantitative detection of porcine epidemic diarrhea viruses. The test strip comprises a sample cushion, a combination cushion, a chromatography film and a water-absorbing cushion, wherein the combination cushion is provided with a fluorescent microsphere labelled anti-PEDV (Porcine Epidemic Diarrhea Viruses) single-domain antibody; the single-domain antibody has high specificity and high sensibility against the antigen PEDV. The test strip is used for detecting PEVD viruses in breeding pig manure or PEDV pollution in feeding stuff plasma proteins on the basis of the newfound immunology principle of the specific single-domain antibody and antigen, can be used for on-site rapid detection or laboratory detection by simply taking 5-10 min; when the test strip is used in combination with a fluorescence quantitative detector, quantitative detection can be realized; the test strip is simple and convenient to operate, operators don't need professional training, a special laboratory is not needed, the limitation of the conventional detection method is overcome, and the test strip has good market prospects.

The invention discloses a kit for time resolution fluorescent quantitative detection on PCT. The kit comprises a fluorescent microsphere antibody complex and an immune fluorescent test paper card, wherein the fluorescent microsphere antibody complex is prepared by marking rare earth fluorescent microsphere with a PCT monoclonal antibody to form a compound and adding the compound on a pipette tip, and is used as a detection antibody after freeze-drying; the immune fluorescent test paper card comprises a detection test paper card; the detection test paper card consists of a sample pad, a nitrocellulose membrane and a water absorbing pad which are adhered to a lining plate in sequence in a lap joint manner; the position of a detection line on the nitrocellulose membrane is wrapped by another PCT monoclonal antibody; the position of a quality control line is wrapped by goat anti-mouse polyclonal antibody. By adopting the kit, the fluorescence influence caused by a sample self can be avoided, the rare earth fluorescent microsphere is adopted as a marking carrier, good stability can be achieved, the microsphere can be connected with the antibody through a covalent bond, and a stable marking product can be generated. The kit is rapid, simple and convenient in detecting the sample and high in sensitivity, and full quantitative detection can be achieved.

The invention discloses a full-automatic immunofluorescence detection device, which comprises a main frame. The main frame is provided with a test operation area, a sample adding area and a detection area. The detection area consists of a reagent card tray transmission mechanism, a scanning mechanism and a temperature controlled incubation mechanism. The sample adding area consists of a sample loading mechanism, a Tip head mechanism and a sample adding mechanism. The detection area consists of an optical detection mechanism and detection software. The scanning mechanism and the temperature controlled incubation mechanism are connected to the detection software, the scanning mechanism is disposed at the upper end of the reagent card tray transmission mechanism, the rear end of the reagent card tray transmission mechanism is equipped with a sample adding platform, reagent cards are sent into the temperature controlled incubation mechanism through a push-card-into-incubation mechanism and an incubation card push mechanism, and the temperature controlled incubation mechanism is provided with a laser scanning mechanism. By adopting the full-automatic immunofluorescence detection device provided by the invention, the detection process is automatically completed mechanically, the efficiency is improved, tedious manual operation is saved, automatic detection is realized, and the detection time is saved. Therefore, the full-automatic immunofluorescence detection device is convenient for application to emergency rooms, laboratory departments and other public places, community hospitals and other occasions, has high cost performance and strong market competitiveness, and fills the domestic blank.

The method comprises: 1) the GoldMag nano-particles are mixed with the double strand DNA; under certain condition, the double strand DNA is fixed on the surface of GoldMag nano-particles, or the GoldMag nano-particles are used to extract the DNA from the human blood, saliva and semen and to fix the extracted DNA on the surface of GoldMag nano-particles; 2) using indirect ELISA method, immumofluorescence method or chemiluminescent method to detect the anti double strand DNA antibody existed in the sampler under test.

The invention relates to a pathogen detection method based on a micro-fluidic chip. The method adopts a micro-fluidic chip integrated with a micro-magnetic field as a reaction container, adopts magnetic balls as solid phase carriers, and adopts quantum dots modified by streptavidin (SA-QDs) as fluorescent markers; under the action of the micro-magnetic field, magnetic balls are captured at specific parts in a chip channel, and thus a micro-reaction zone is formed; pathogens are captured through a sandwich immunization reaction; with the interaction between biotin and streptavidin, the immunofluorescence quantitative analysis of the pathogens is realized. The detection method combines the micro-fluidic chip, magnetic immunoassay, and fluorescence detection, has the advantages of rapidness, high efficiency, and few sample using amount for the micro-fluidic chip, high specificity and strong manoeuvrability for magnetic immunoassay, and excellent optical properties for quantum dots, and is a multi-objective real-time pathogen detection method.

The invention discloses an improved immune fluorescyte staining method, comprising 5 steps: fixing/permeating, permeating, surface staining, staining in nucleus, washing and weight dropping; the method of the invention is an improvement to the Foxp3 immune fluorescyte staining method of eBioscience Company, changes a two-step multiple staining method into a one-step multiple staining method, and carries out the staining on the surface of the cell and in the nucleus simultaneously, thus having the advantages of simple and convenient operation, quick staining, saving reagent, and reducing the cost, etc. The research shows that the method of the invention has no remarkable difference with the method of that of the eBioscience Company, can be used for the staining on the surface of the cell and in the nucleus simultaneously and has extremely good application prospect and promotional value.