97 results about "Fluorescent labelling" patented technology

This invention describes systems, primers, kits, and methods for detecting microsatellite instabilities in biological samples. Signal data is received from a capillary electrophoresis genetic analysis instrument, wherein the signal data is determined based on the fluorescence of fragments comprising nucleic acid sequences amplified from the biological sample by polymerase chain reaction (PCR). The nucleic acid sequences correspond to multiple different microsatellite loci and are obtained using multiple PCR primers configured to side-join the multiple microsatellite loci of the biological sample. When the PCR primers are combined with the biological sample and subjected to PCR amplification, fluorescently labeled DNA fragments comprising the multiple microsatellite loci are generated. The microsatellite instabilities of the biological sample are classified using the fluorescence data obtained from the multiple fluorescently labeled microsatellite loci.

This invention provides a LYTACA polypeptide complex (sRD-Lytaca) targeting the degradation of soluble IL-17RD (sIL-17RD), its preparation method, and its applications. This polypeptide complex is formed by the self-assembly of the sIL-17RD-binding peptide sRDL and the scavenger receptor-binding peptide SRAL after Nap-FF modification and mixing. sRDL can be conjugated with fluorescently labeled molecules to prepare detection reagents for the detection of sIL-17RD. This invention also provides a variant with at least 80% sequence identity with sRDL and retaining binding ability, and pharmaceutical compositions containing sRDL and / or sRD-Lytaca. This invention provides a new technical means for the prevention and treatment of osteoarthritis and diseases related to sIL-17RD protein (such as rheumatoid arthritis).

This invention relates to the field of cell detection technology, specifically to a reagent tube for detecting mesenchymal stem cell phenotypes. The tube includes a reagent tube body and pre-encapsulated fluorescent antibody lyophilized microspheres. The lyophilized microspheres are formed by lyophilization of a mixture of lyophilization protection solution and fluorescently labeled antibody. The fluorescently labeled antibody is a combination of antibodies targeting positive markers of mesenchymal stem cell phenotypes CD73, CD90, and CD105, or a combination of antibodies targeting negative markers CD45, CD34, CD11b, CD19, and HLA-DR. This invention offers high efficiency and convenience: by pre-encapsulating the fluorescent antibody lyophilized microspheres, the traditional three-step process of "antibody preparation-sample addition-mixing" is simplified to direct addition of cell suspension. The single-sample detection time is reduced from 120 minutes to 30 minutes, the number of pipetting operations is reduced by 80%, and human error is significantly reduced.

This invention provides a simple method for evaluating whether or not a peptide binds to isovaleric acid. [Solution] A method for evaluating the binding affinity to isovaleric acid, comprising a preparation step of preparing a fluorescently labeled compound in which a fluorescent group is bonded to the α-carbon atom of isovaleric acid, and a contact step of contacting the fluorescently labeled compound with a peptide fragment of an olfactory receptor.

This invention relates to DNA-functionalized thermosensitive phase transition polymers, their preparation methods, and applications, belonging to the interdisciplinary field of functional polymers and biomaterials. It aims to solve the technical problems of existing exosome separation methods, such as long processing time, low purity, high cost, and inability to specifically separate exosome subtypes. This invention provides a polymer prepared via free radical polymerization using N-isopropylacrylamide and acrylamide-modified DNA as monomers, and ammonium persulfate and tetramethylethylenediamine as initiators. This polymer exhibits reversible phase transition properties at physiological temperatures and is applied to the separation and detection of exosome subtypes. Specific capture and quantitative analysis are achieved through DNA recognition probes and fluorescently labeled antibodies. The polymer of this invention is simple to operate, quick to prepare, low in cost, and high in purity. It requires only a common centrifuge for non-destructive separation of exosomes and can integrate detection functions, providing technical support for diagnosis and detection.

A continuous analysis method comprising generating microfluidic droplets comprising multiple copies of a same antigen, multiple copies of a same primary antibody potentially capable of specifically binding to said antigen, a multivalent crosslinking agent being a biomolecule or a support, and a fluorescent labeling agent, said continuous microfluidic analysis being a fluorescence-based sorting.

The application relates to a kind of lipid nanocarriers coupled with GO-203 and loaded with DMAMCL and its preparation method and application, belong to the field of biological medicine and nano-preparation.The lipid nanocarriers include water phase core, lipid bilayer and surface modification layer from inside to outside, wherein water-soluble dimethylamine methyl chloride (DMAMCL) is loaded in the water phase core;The lipid bilayer is wrapped in the water phase core, and coumarin-6 is embedded as fluorescent marker for tracing and detection;Functional modification layer is arranged on the outer surface of the lipid bilayer, including DSPE-PEG2000-COOH and DSPE-PEG2000-GO-203, wherein GO-203 is cell membrane penetrating peptide.The lipid nanocarrier provided by the application can realize the synergistic delivery of GO-203 and DMAMCL, be used for regulating tumor immune microenvironment and be used for the treatment of triple-negative breast cancer.

The application discloses a circRNA detection method based on Argonaute protein and rolling circle amplification. The method comprises the following steps: mixing a sample containing target circRNA to be detected with a specific reverse transcription primer and reverse transcriptase to obtain long-chain cDNA; mixing the obtained long-chain cDNA with pfAgo, gDNA and a DNA nucleic acid probe to carry out a reaction, and calculating the content of the target circRNA according to the fluorescence signal of the reaction mixture; the nucleotide sequence of the gDNA is specifically complementary to the cDNA sequence obtained by reverse transcription of the circRNA; the sequence of the DNA nucleic acid probe is specifically complementary to the sequence of the cDNA after one cutting of the cDNA by the pfAgo; and the DNA nucleic acid probe is a single-stranded DNA probe with a fluorescent label. The application can realize high-sensitivity, high-specificity and multiple target simultaneous analysis and detection of potential diagnostic biomarkers of liver cancer such as hsa_circ_0001445 and hsa_circ_0001141.

This invention discloses a single-cell microarray-based kit and method for in situ quantitative detection of receptor-binding allergen-specific IgE. The kit includes: a single-cell microcavity array chip, an antigen barcode chip, a plastic clamp, IgE standards, fluorescently modified IgE detection antibodies, and auxiliary reagents. During detection, basophil single cells are loaded into the microcavity array chip, lysis buffer is added, and the cells are clamped together with the antigen barcode chip. Low-temperature incubation allows the allergen-specific IgE released from cell lysis to be captured in situ by the antigen on the chip. A sandwich immunoassay is then performed using a fluorescently labeled antibody, and absolute quantification is achieved using a standard curve. This invention is the first to achieve high-throughput, high-sensitivity, multi-component parallel in situ quantitative detection of basophil membrane receptor-binding sIgE at the single-cell level. It can directly reflect the functional sensitization state of effector cells and has advantages such as simple operation, low sample requirement, and no need for complex valve devices.

This disclosure provides immunoassay reagent compositions with increased stability (when exposed to light). These compositions typically include dyes that prevent degradation of labels, such as fluorescent labels. Kits comprising these compositions, methods of use, and methods of manufacture are also provided.

This invention belongs to the field of genetic engineering technology, specifically disclosing a method for preparing and applying gene-modified homozygous cells of chicken progenitor cells (PGCs). Using a dual-fluorescent marker gene, two donor plasmids carrying green fluorescent protein (EGFP) and red fluorescent protein (mCherry), respectively, are transfected into PGCs. Cells simultaneously expressing red and green fluorescence are obtained using flow cytometry, thus forming homozygous cell lines. Then, the dual-fluorescent marker gene is deleted using the Cre-LoxP system combined with electroporation, and residual LoxP fragments are deleted using ssODN as a donor template, achieving traceless gene editing in recipient chicken embryos. This invention does not introduce any exogenous genes, increasing the safety of gene-modified chickens and is of great significance for eliminating the biosafety risks associated with gene-edited chickens.

The invention provides a high-flux monosaccharide separation and quantification method and kit based on gel electrophoresis. The separation and quantification method comprises the following steps: (1) providing a fluorescence-labeled monosaccharide sample to be detected; (2) injecting the monosaccharide sample to be detected into a capillary electrophoresis system, and performing first electrophoretic separation under a first separation system to obtain a first electrophoretogram; the first separation system comprises borate-free first separation gel and a first buffer solution; (3) cleaning a capillary tube of the capillary electrophoresis system, injecting the monosaccharide sample to be detected into the capillary tube again, and performing electrophoretic separation for the second time in a second separation system to obtain a second electrophoretogram; the second separation system comprises second separation gel containing borate and a second buffer solution; and (4) carrying out qualitative identification and quantitative calculation on the mixed monosaccharide according to the migration time difference and peak overlapping condition of each monosaccharide component in the first electrophoretogram and the second electrophoretogram.

An in vitro diagnostic assay method, comprising: providing a sample which may comprise target analyte(s); contacting the sample with a fluorescent label whose fluorescence can be externally modulated, such that the fluorescent label is associated with target analyte(s), if present, to form a fluorescent label-analyte complex; and detecting the fluorescent label-analyte complex; and device, kit and solid phase for performing an in vitro diagnostic assay method.

The present application belongs to the technical field of intelligent coating, and relates to a composite coating material with fluorescent self-indication function for micro stress detection and a preparation method and application thereof. The present application realizes high sensitivity detection of micro cracks by adding fluorescently labeled microcapsules capable of breaking at the crack and releasing fluorescent dyes at the fixed point, and effectively avoids the adverse effects of microcapsule introduction on the transparency and flexibility of the coating through refractive index matching, particle size optimization and film thickness design, thereby being suitable for early warning of corrosion risk of various substrates, providing sufficient window period for equipment maintenance and coating repair, and significantly improving the safety and reliability of the protective coating.

The application provides a kit and method for detecting common fusion genes of lymphoid leukemia based on a multiple digital PCR method. By constructing a synergistic mechanism of primers and fluorescent probes, optimizing the collocation of primers, probes and fluorescent markers, and effectively avoiding the non-specific binding between amplification products. The innovative introduction of a composite reaction aid with a specific formula, namely a PCR enhancer, significantly improves the amplification efficiency and specificity of the multiple detection system. The kit provided by the application realizes the integrated detection capability of seven fluorescent channels, Atto 425, VIC, FAM, ROX, CY5, CY5.5 and CY7 in a single hole design. By innovatively constructing a ROX+CY5 and Atto 425+VIC dual-channel combined detection system, when the ROX and CY5 channels or the Atto 425 and VIC channels in the same detection hole simultaneously present positive signals, two additional detection targets can be specifically interpreted. This innovative detection strategy based on channel combination enables the kit to break through and achieve accurate detection of 18 fusion genes with only 2 detection holes.

This invention belongs to the field of nucleic acid aptamer technology. This invention provides a high-affinity and specific phalloidin nucleic acid aptamer and its applications. The base sequence of the high-affinity and specific phalloidin nucleic acid aptamer is shown in SEQ.ID.NO.1. The nucleic acid aptamer obtained by this invention exhibits strong specificity for phalloidin; after detection by graphene oxide fluorescence quenching, the measured fluorescence intensity reaches 867.6 (au), while its effects on other toxins and blank controls are not significant. Furthermore, it has high affinity for phalloidin, with a Kd value of 240 nM (0.203 ng / Kg). After 5' end FAM fluorescent labeling, the nucleic acid aptamer obtained by this invention can achieve visual detection of phalloidin under ultraviolet transmission irradiation.

A method of controlling the concentration of a corrosion inhibitor in closed loop cooling water includes feeding a cooling water mixture containing a corrosion inhibiting composition including at least one copper corrosion inhibiting compound and at least one tagged polymer including a fluorescent moiety in the backbone thereof, through a circulating loop that circulates the cooling water past a copper surface, and on a sample of the cooling water mixture downstream from the copper surface in the circulating loop, measuring a fluorescence signal of the sample to determine a circulating concentration of the composition in the cooling water mixture and determine what, if any, adjustment is required in concentration.

The present disclosure relates to an information processing system and an information processing method using a server system and a data analysis client terminal. The main object is to provide an information processing system capable of coping with an increase in the amount of information processing involved in the analysis of biological samples. The present disclosure provides a server system including: an automatic analysis processing unit that generates output data by performing analysis processing on light intensity data or fluorescent marker intensity data obtained by irradiating light on a biological sample; an analysis result data storage unit that stores output data generated based on the light intensity data or the fluorescent marker intensity data; and an interactive analysis processing unit that analyzes the fluorescent marker intensity data based on an analysis command related to the output data output to an output device, and outputs analysis result data. The present disclosure also provides an information processing system including the server system.

The application discloses a near-infrared two-region nano fluorescent microsphere, a method for preparing a test strip of the near-infrared two-region nano fluorescent microsphere and labeled antibodies, a near-infrared two-region fluorescent detection system nano fluorescent microsphere and a preparation method thereof, and a near-infrared two-region fluorescent labeled antibody and a preparation method thereof. The nano fluorescent microsphere comprises a polystyrene substrate and near-infrared two-region fluorescent molecules with an aggregation-induced emission characteristic loaded in the polystyrene substrate. The near-infrared two-region fluorescent molecules have the following general structure: wherein R1 is independently selected from one of H, -CH3 and -OCH3, R2 is independently selected from H, R3 is independently selected from H or an electron-donating group containing a triphenylamine derivative. The nano fluorescent microsphere provided by the application has a fluorescent quantum yield of 5.4% or more, which is significantly higher than that of an existing NIR-II fluorescent probe. The fluorescent intensity retention rate is more than 90% after 60 minutes of irradiation. The NIR-II waveband has extremely low background interference, and the signal-to-noise ratio is significantly better than that of a visible light region fluorescent immunochromatography method.

The present application relates to the technical field of biological medicine, in particular to a kind of nano-molecular probe with targeting and its preparation method and application.The plasmid of immune checkpoint corresponding receptor of stable expression tumor surface negative regulation molecule is synthesized in the present application, which is prepared into targeted nano-molecular probe with cell membrane nanocapsule of enhancing T cell activity ligand and being marked with fluorescence, and melanin nanosphere of simultaneously loading therapeutic drug.The targeted nano-molecular probe provided in the present application can be long-circulated in vivo, reduce the dosage and administration frequency of chemotherapeutic drug.It also has the advantages of improving the drug resistance of tumor site to therapeutic drug, targeting tumor DNA damage and significantly enhancing the infiltration and activation of immune cells in tumor site.

The purpose of this invention is to provide a method for detecting mutant genes that can comprehensively and without omission measure the mutation rate of genes while suppressing costs. The method for detecting mutant genes of this invention involves performing a base extension reaction using a first pigment group and a second pigment group, respectively. The first pigment group is configured such that one of the bases A and B is not fluorescently labeled, while the other is fluorescently labeled, and GC is fluorescently labeled. The second pigment group is configured such that one of the bases C is not fluorescently labeled, while the other is fluorescently labeled, and AT is fluorescently labeled. Both the first and second pigment groups are configured such that the base extension reaction does not stop at the unlabeled bases but stops at the fluorescently labeled bases (see Figure 6).

The present application relates to the technical field of aquatic animal cell culture, and particularly relates to a rapid enrichment and separation method of Plectrypops leopardus gonadal somatic cells. The method comprises the following steps: selecting healthy Plectrypops leopardus for anesthesia; taking gonadal tissue, and after aseptic treatment, cutting, centrifugal washing and filtration, inoculating in a culture medium for primary culture to obtain gonadal cells migrated from the tissue block, and after trypsin-EDTA digestion and centrifugal washing, using 1xPBS containing polysorbate for treatment; after blocking, antibody incubation and washing, verifying the labeling effect by a fluorescence microscope to realize specific enrichment of gonadal somatic cells; after filtration and purification of the enriched fluorescent labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescent positive cells under a fluorescence field, and transferring the cells to a culture dish containing complete culture medium, inoculating the separated fluorescent positive cells in optimized complete culture medium for purification culture, so that the cells adhere to the wall and realize stable subculture.

The present application relates to the technical field of sex chromosome STR identification, and particularly relates to an X chromosome STR locus composite amplification primer group, a 9-color fluorescent label composite amplification kit and application thereof.The composite amplification primer group comprises 45 pairs of specific amplification primer pairs, the 45 pairs of specific amplification primer pairs include 44 X chromosome STR loci and 1 pair of specific amplification primer pairs of a gender locus AMEL, and the nucleotide sequences of the primer pairs in the composite amplification primer group are shown as SEQ ID NO.1-90.The specificity of each primer in the composite amplification primer group is relatively strong, no primer dimer or interaction is generated during composite PCR amplification, the amplification efficiency difference of each pair of primers is relatively small, and the identification sensitivity is relatively high.The composite amplification primer group provides a molecular basis and core necessary reagent for simultaneously detecting 44 X-STR loci at a time.

This application discloses a method for screening single B cells. The method provided in this application uses magnetic beads coated with IL-6 antigen, combined with fluorescently labeled secondary antibody, and employs droplet microfluidic technology to rapidly sort B cells. The obtained IL-6 positive B cells are then directly cultured, and the generated antibodies are subjected to ELISA and IL-6 antigen-antibody affinity detection to obtain the target cell line. The entire process is greatly simplified and the workload is significantly reduced.

The application belongs to the technical field of nucleic acid detection, and particularly relates to a CRISPR cascade nucleic acid detection system based on a DNA signal amplification reactor, a preparation method and application thereof, which comprises a DNA signal amplification reactor, a Cas13a recognition module and a Cas12a signal output module, the DNA signal amplification reactor is internally packaged with a fluorescently labeled probe and a Cas12a activation substrate; the Cas13a recognition module is used for triggering cutting of a connecting chain after recognizing a target RNA, and releasing the DNA signal amplification reactor; the Cas12a signal output module is used for recognizing the released DNA signal amplification reactor and generating a fluorescent signal, the Cas12a signal output module activates trans-cleavage activity after recognizing the released DNA signal amplification reactor, cuts a fluorescent reporter probe, and realizes double-stage signal amplification, the application combines the DNA reactor with a double CRISPR / Cas cascade system for non-amplification detection of pathogens RNA. The application is suitable for high-sensitivity, high-specificity and non-amplification instant detection of pathogens RNA.

The present application relates to a tunable annular diaphragm cell imaging system and a dye-free deep learning identification method, relating to the technical field of optical imaging and image processing, comprising an annular diaphragm illumination module; the present application establishes a quantitative mapping relationship between the outer diameter of the annular diaphragm parameter and the image quality index, and adaptively finds the best bright field and dark field illumination conditions for different cell types, densities and overall profiles and internal structures, so as to obtain an original image with sharp edges and distinct internal structures without using any dye, the improved deep learning segmentation model has a two-way attention gate module, which can capture spatial details and texture semantic information important for cell recognition in parallel and differentially, realize high-accuracy dye-free cell dead or alive automatic identification, make the segmentation result more consistent with the biophysical characteristics of cells, improve the segmentation accuracy and recognition accuracy, and automatically distinguish the state of cells with complex background without fluorescent labeling.

The application belongs to the technical field of plastics, and particularly relates to a plastic particle performance detection process in an injection molding machine, which aims at the problem that existing detection data can only be obtained and material conditions in the detection process cannot be understood, and the following scheme is proposed, which comprises the following steps: S1: obtaining plastic particles, and then obtaining a marker material, the amount of the marker material being 0.2% to 1% of the amount of the plastic particles, and the marker material comprising fluorescent powder, indolizine, powdered rubber, ethylene glycol and paraffin wax; S2: pouring the plastic particles and the marker material into a stirring device for stirring, and performing fluorescent marking on the material of the plastic particles by the marker material, so that a fluorescent picture of the plastic particles is obtained in the performance detection process, the change of the plastic particles before and after the detection and in the detection process is understood by using the fluorescent picture, and the change of the material attached with the fluorescent is understood, and if the fluorescent gathers together or the fluorescent is unevenly distributed, it is indicated that the plastic particles are not uniformly stirred in the production process.