64 results about "Fluorescent cell" patented technology

The present invention provides novel fluorescent compounds and covalent attachment chemistries which facilitate the use of these compounds as labels for ultrasensitive and quantitative fluorescent detection of low levels of biomolecules. In a preferred embodiment, the fluorescent labels of this invention are novel derivatives of the hydroxy-pyrene trisulphonic and disulphonic acids which may be used in any assay in which radioisotopes, colored dyes or other fluorescent molecules are currently used. Thus, for example, any assay using labeled antibodies, proteins, oligonucleotides or lipids, including fluorescent cell sorting, fluorescence microscopy (including dark-field microscopy), fluorescence polarization assays, ligand, receptor binding assays, receptor activation assays and diagnostic assays can benefit from use of the compounds disclosed herein.

The invention provides a high content image flow biological microscopic analysis system. The system comprises a liquid flow system and an optical system, wherein the liquid flow system enables cells in sample cell suspension to be focused in the center of a liquid flow under the constraint of the system sheath liquid flow and to flow through a detection window of a flow chamber one by one; the optical system comprises optical sources, an optical path system and an optical filter stack; the optical sources include a halogen lamp and a laser; through the optical path system and the optical filter stack formed by a plurality of dichroscopes, the optical system generates a bright field cell image, a six-channel fluorescent cell image and six PMT (photomultiplier tube) optical intensity signals with different wave bands. The high content image flow biological microscopic analysis system integrates flow cytometry and fluorescence microscopic imaging, has a plurality of detection channels, can collect the cell images of the cells passing through the flow chamber one by one and can carry out quantitative analysis on each cell image by utilizing analysis software, thus providing the statistical data of a cell group and the information of the cell morphology, cell structures and subcellular signal distribution.

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.

The present invention is directed to a method of separating oligodendrocyte cells or progenitor cells thereof from a mixed population of cells. It comprises selecting a promoter which functions only in the oligodendrocyte cells or progenitor cells thereof, introducing a nucleic acid molecule encoding a fluorescent protein under control of that promoter into the mixed population cells, allowing the oligodendrocyte cells or progenitor cells thereof to express the fluorescent protein, and separating the fluorescent cells from the mixed population cells, where the separated cells are the oligodendrocyte cells or progenitor cells thereof. The invention also relates to the isolated and enriched human oligodendrocyte cells or progenitor cells thereof.

The invention provides a fluorescent cell automatic counting method and device, equipment and a medium. The method comprises the following steps: S1, reading a microscopic image of fluorescent cells;S2, preprocessing the microscopic image of the fluorescent cell, including correcting uneven illumination of the microscopic image and enhancing the contrast between the cell in the microscopic imageand the background; adopting median filtering to filter microscopic image noise, and reserving detail information of the edge of the image; removing microscopic image noise and background impurities by adopting opening operation, smoothing cell contours, disconnecting cell stenosis connection and removing small protruding parts of cells; s3, segmenting the fluorescent cells adhered to the microscopic image to prevent the adhered cell population from serving as one cell during cell counting; and S4, calculating the number of the fluorescent cells. Requirements of most laboratories can be met from the aspects of efficiency, accuracy and cost.

The present invention provides novel fluorescent compounds and covalent attachment chemistries which facilitate the use of these compounds as labels for ultrasensitive and quantitative flourescent detection of low levels of biomolecules. In a preferred embodiment, the flourescent labels of this invention are novel derivatives of the hydroxy-pyrene trisulphonic and disulphonic acids which may be used in any assay in which radioisotopes, colored dyes or other fluorescent molecules are currently used. Thus, for example, any assay using labeled antibodies, proteins, oligonucleotides or lipids, including fluorescent cell sorting, fluorescence microscopy (including dark-field microscopy), fluorescence polarization assays, ligand, receptor binding assays, receptor activation assays and diagnostic assays can benefit from use of the compounds disclosed herein.

The invention provides a high content image flow biological microscopic analysis method. The method comprises the following steps: building a high content image flow biological microscopic analysis system, and detecting cells by utilizing the high content image flow biological microscopic analysis system, wherein in the high content image flow biological microscopic analysis system, a liquid flow system enables cells to be focused in the center of a liquid flow and to flow through a detection window of a flow chamber one by one; an optical system comprises optical sources, an optical path system and an optical filter stack; the optical sources include a halogen lamp and a laser; through the optical path system and the optical filter stack formed by a plurality of dichroscopes, the optical system generates a bright field cell image, a six-channel fluorescent cell image and six PMT (photomultiplier tube) optical intensity signals with different wave bands. The method has the beneficial effects that the high content image flow biological microscopic analysis system integrates flow cytometry and fluorescence microscopic imaging, has a plurality of detection channels, can collect the cell images of the cells passing through the flow chamber one by one and can carry out quantitative analysis on each cell image by utilizing analysis software, thus providing the statistical data of a cell group and the information of the cell morphology, cell structures and subcellular signal distribution.

The invention belongs to the technical field of cell marking and particularly relates to a GFP-CD19 fusion protein, a preparation method and application of the GFP-CD19 fusion protein in the cell marking aspect. The GFP-CD19 fusion protein comprises 776 amino acid, and the molecular weight is about 85 KD; the fusion protein is used for marking the cells with anti-CD19 bodies expressed on the surfaces. A specific combination principle of antigen-antibody or receptor-ligand is used, the GFP-CD19 fusion protein is specifically prepared, the protein can be specifically combined with a chimeric antigen receptor and is subjected to fluorescent marking, only T-lymphocyte which is effectively transfected and embedded with an antigen receptor can be combined with the protein so that the number of the effectively transfected T cells can be calculated by detecting the number of fluorescent cells, the number of effective T-lymphocyte returned to a patient body can be accurately controlled according to the data, and accordingly, guarantee is provided for treating the tumor with the CAR-T technology.

The invention discloses a method for applying calixarene fluorescent probes to fluorescent imaging of Zn<2+> and F<->. Two thiacalix[4]arene fluorescent probes selectively bonded with Zn<2+> and F<-> and having chemical names of 1,3-alternative-5,11,17,23-tetra-tert-butyl-25,27-di[(7-hydroxy-8-coumarinimino)ethoxy]-26,28-di(2-methoxyethoxy)thiacalix[4] and 1,3-alternative-5,11,17,23-tetra-tert-butyl-25,27-di[(7-hydroxy-8-coumarinimino)ethoxy]-26,28-thiacalix[4]crown-5 are respectively used as fluorescent imaging reagents for trace Zn<2+> and F<-> in active cancer cells in the human body; and the probes are compatible with the active cells and have permeability and nontoxicity. After the probes 1 and 2 are used to respectively dye Zn<2+> and F<-> in the cells, an inverted fluorescence microscope is used to detect shot clear blue fluorescent cell images.

The present invention relates to a method of separating multipotential neural progenitor cells from a mixed population of cell types. This method includes selecting a promoter which functions selectively in the neural progenitor cells, introducing a nucleic acid molecule encoding a fluorescent protein under control of said promoter into all cell types of the mixed population of cell types, allowing only the neural progenitor cells, but not other cell types, within the mixed population to express said fluorescent protein, identifying cells of the mixed population of cell types that are fluorescent, which are restricted to the neural progenitor cells, and separating the fluorescent cells from the mixed population of cell types, wherein the separated cells are restricted to the neural progenitor cells. The present invention also relates to an isolated human musashi promoter and an enriched preparation of isolated multipotential neural progenitor cells.

The invention relates to an NO<3><-> ion detection reagent and an application thereof, and belongs to the technical field of analysis chemistry. A tiny amount of NO<3><-> in a neutral aqueous solution is detected through a fluorescence method by using a 9,10-dipyridylvinyl anthracene cation derivative as a fluorescence probe a. When NO<3><-> in the aqueous solution is determined, the fluorescence intensity at 575nm is determined by using 470nm as fluorescence excitation wavelength, the detection concentration linear range is 1.5*10<-7>-2.1*10<-5>mol.L<-1>, and the lowest detection limit is 10<-8>mol.L<-1>. After the probe a and NO<3><-> are intracellularly stained in a living cell imaging, a fluorescence inverted microscope detection result shows that clear yellow fluorescence cell distribution exists, a cell image is shot to obtain a clear cell contour image, and staining has specific area selective aggregation capability.

The invention relates to a preparation method of triple responsive nanogel and application of the triple responsive nanogel, which belong to the technical field of macromolecular materials, and relate to preparation and application of polymer nanogel. The preparation method comprises the following steps: adding spiropyrane methacrylate prepared by virtue of an acylation reaction, acrylic acid, N,N-bis(acrolyl) cystamine and lauryl sodium sulfate according to a molar ratio of 2: 30: 1: (1 to 2): 50: 1: 1 into a container fully filled with nitrogen, reacting for a plurality of hours at a certain temperature, and freeze drying to obtain the triple responsive polyacrylic acid-co-spiropyrane methacrylate nanogel. By adopting the preparation method, the carrying of the nanogel for guest molecules is realized, and the controlled release under different single or cooperative out-field irritation of light, pH and a reduction substance (DTT) is realized; and meanwhile, the preparation method is simple, lowcost, capable of being well adaptive to complicated living body environment and capable of being used for biological fluorescent cell imaging treatment and drug delivery.

The embodiment of the invention discloses a portable fluorescent cell analysis system and a microscopic imaging method thereof. The portable fluorescent cell analysis system comprises a microscopic imaging module, an objective table and a light source; the microscopic imaging module comprises an image acquisition unit and a microscopic amplification unit, and the effective working distance L1 between the image acquisition unit and the objective table meets the condition that L1 is larger than or equal to 30 mm and smaller than or equal to 100 mm in the direction perpendicular to the objectivetable; wherein the microscopic amplification unit at least comprises an amplification objective lens group, the amplification objective lens group comprises at least one objective lens, and the focallength f of the objective lens is greater than or equal to 2mm and less than or equal to 10mm; the light source comprises a bright field light source and at least one fluorescent light source. By theadoption of the technical scheme, it is guaranteed that fluorescence microscopic imaging can be achieved, it is guaranteed that the microscopic imaging module is small and compact by reasonably setting the effective working distance between the image acquisition unit and the objective table and reasonably setting the focal length of the objective lens, and the integration degree and portability ofthe portable fluorescence cell analysis system are improved.

The invention belongs to the technical field of bioengineering and protein activity detecting and discloses a recombinant expression vector pEGFP (Plasmid Enhanced Green Florescence Protein)-DsRed-BP and a preparation method thereof. The expression vector contains a vector skeleton pEGFP-C1, a reverse complement inserted DsRed gene sequence, a phiC31 integrase identification sequence attB and a reverse complement inserted attP sequence. The invention further discloses a method for detecting the activity of phiC31 integrase in a mammalian cell by adopting the expression vector. The method is used for quantitatively or quantitatively detecting the activity of the phiC31 integrase in the mammalian cell according to the type of fluorescence emitted by a co-transfected cell or the proportion among different fluorescent cells. According to the method, the activity of the phiC31 integrase in the mammalian cell can be intuitively evaluated through fluorescent protein just by once transfection, and therefore, the detection method is simple and convenient, reliable in result and low in detection cost.

The invention discloses rare earth nanoparticles with intense red fluorescence, a preparation method of the rare earth nanoparticles and an application of the rare earth nanoparticles to cell imaging.The rare earth nanoparticles comprise rare earth europium ions, terbium ions and carbon quantum dots, have the particle size being smaller than 5 nm, and emit characteristic fluorescence of the europium ions. According to the dual energy transfer effect of the carbon quantum dots and the terbium ions, the rare earth nanoparticles can glow strongly in an aqueous solution without organic ligands, so that the defects that a conventional fluorescence imaging material containing the organic ligands requires complex organic synthesis and needs an organic solvent to help solubilization due to poor water solubility are overcome. The rare earth nanoparticles are low in cytotoxicity, have good biocompatibility and safety and can be applied to fluorescent cell imaging.

The invention provides a fluorescent cell model for screening an M2 ion channel blocker for influenza virus, and the cell model is a cell containing pH sensitive fluorescent materials. Meanwhile, the invention also provides a method for screening the M2 ion channel blocker for the influenza virus by the cell model, which is to detect the change of fluorescent values in the cell model under the condition of an acidic buffer. The invention has the advantages that the H+ flowing condition through the M2 ion channel is directly reflected by detecting the change of the fluorescent values reflecting pH in the cell; and the M2 ion channel blocker screening method established by the cell model has definite mechanism of drug action, is simple and convenient, quick and direct, and is suitable for high-flux accurate screening of the M2 ion channel blocker.

The invention discloses a fluorescence-patch clamp-micropipette detection device. An experimental accommodating cavity, a three-dimensional micromanipulator and a patch clamp are arranged on an experiment platform, and two sides of the experimental accommodating cavity are hollowed out for a micropipette and a glass electrode to enter; fluorescent cells are contained in the experimental accommodating cavity, the micropipette sucks red blood cells, the fluorescent cells and the red blood cells are positioned in an extracellular fluid in the experimental accommodating cavity, and the glass electrode is filled with an electrode fluid; the patch clamp comprises a patch clamp amplifier, a patch clamp probe and a patch clamp probe bracket; full-spectrum light emitted from the fluorescence lightsource passes through a color filter to form fluorescence incident light, the fluorescence incident light enters the cells in the experimental accommodating cavity, and fluorescence of the cells returns to be received by means of a fluorescence camera. According to the fluorescence-patch clamp-micropipette detection device, the influence of the cell membrane potential change on the membrane protein interaction can be detected while a high signal-to-noise ratio fluorescence image is collected to research transmembrane signal transduction of the membrane protein, and the coupling relationship among the three spectrum phase information can be recorded while the fluorescence spectrum, the electrophysiological spectrum, the adhesion state spectrum and other three spectrum phase information aresynchronously recorded.

A process of in vivo labeling and identifying recombinantly produced peptides or proteins within an unpermeabilized prokaryotic host cell. Recombinant prokaryotic cells expressing a fusion peptide comprising at least one tetracysteine tag were labeled in vivo using a biarsenical labeling reagent. A fluorescent activated cell sorter was used to identify and select subpopulations of fluorescent cells wherein the amount of fusion peptide in the cell was proportional to the amount of fluorescence detected.

The invention discloses a fluorescent probe method for monitoring Zn<2+> in cancer cells. The fluorescent probe method utilizes bis-(7-hydroxy-coumarin-8-aldehyde)ethylenediamine as a fluorescent probe of a trace amount of Zn<2+>. Through the selectively-formed probe-Zn<2+> complex characteristic of emitting 457nm wavelength strong fluorescence, the probe and Zn<2+> are respectively permeated into active cancer cells so that fusion with the cells is realized. After Zn<2+> in cells is dyed by the probe, a distinct blue fluorescent cell image is detected and showed by an inverted fluorescence microscope. The probe is used as a fluorescent imaging reagent of Zn<2+> in active cancer cells. The probe has a chemical structural formula shown in the following description.

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.

Provided is a 2,7-bis(4-vinylpyridine)-9-fluorenone salt two-photon fluorescent cell nucleus positioning probe material. The structural formula of the positioning probe material is shown in the specification, wherein R represents CH3 or C2H5 or C4H9. A preparation method of the positioning probe material comprises the steps that reactants such as 2,7-dibromo-9-fluorenone and 4-vinylpyridine are put in a mixture which takes palladium acetate as a catalyst, adopts anhydrous potassium carbonate to provide an alkaline environment and takes N-methyl pyrrolidinone as solvent, a heck reaction is conducted in a sealed container in a nitrogen protection mode under the conditions of certain temperature and pressure, and then a final product is obtained after iodoalkane salinization is conducted. According to the 2,7-bis(4-vinylpyridine)-9-fluorenone salt two-photon fluorescent cell nucleus positioning probe material and the preparation method and application thereof, long-time real-time continuous observation capacity is embodied in cell nucleus dyeing marking and two-photon fluorescent imaging, a biological optical window (800-860 nm), located in a near-infrared band, of light is excited, and Rayleigh scattering of exciting light is effectively reduced. The 2,7-bis(4-vinylpyridine)-9-fluorenone salt two-photon fluorescent cell nucleus positioning probe material can be applied to the field of two-photon three-dimensional bioluminescent imaging.

The present invention relates to an enriched or purified preparation of isolated hippocampal neural progenitor cells and progeny thereof. The present invention also relates to a method of separating neural progenitor cells from a mixed population of cell types from hippocampal tissue. This method includes selecting a promoter which functions selectively in the neural progenitor cells, introducing a nucleic acid molecule encoding a fluorescent protein under control of said promoter into all cell types of the mixed population of cell types from hippocampal tissue, allowing only the neural progenitor cells, but not other cell types, within the mixed population to express said fluorescent protein, identifying cells of the mixed population of cell types that are fluorescent, which are restricted to the neural progenitor cells, and separating the fluorescent cells from the mixed population of cell types, wherein the separated cells are restricted to the neural progenitor cells.

The invention provides a method for obtaining the number and area of cells based on a multicolor fluorescent picture, which is characterized by comprising the following steps of: obtaining the multicolor fluorescent picture; separately processing the three pictures obtained through separation, and obtaining the cell number and the cell area of a single picture; and obtaining the cell number and the cell area of the three pictures obtained by separating the current multicolor fluorescent picture, and carrying out partition counting on all the cell areas according to the size, so that the cell number and the cell area of the current multicolor fluorescent picture are obtained after the cell number and the cell area of the three pictures are combined. The method for obtaining the number of fluorescent cells provided by the invention can be used for accurately analyzing the number and size of cells in a fluorescent picture in batches, so that the problems of low counting and area statistical efficiency and accuracy, complicated operation and the like of the existing multi-color fluorescent staining cell counting methods such as immunofluorescence, TUNEL, EdU and the like are solved.

Methods are provided to facilitate the detection and enumeration of neutrophils in bodily fluids, including milk. The methods incorporate the fluorescent staining of neutrophils using fluorogenic enzyme substrates, the imaging of fluorescent cells using a digital device, and the electronic counting of the cells.

The invention discloses a fluorescent cell sensor for screening inflammasome NLRP3 activators and inhibitors, and belongs to the technical field of drug screening. The cell fluorescence sensor is constructed by transgenic means. An NLRP3 promoter core region and a ZSGREEN gene are inserted into a plasmid vector and is transferred into Thp-1 cells to obtain a stable cell line; the stable cell line emits green fluorescence after receiving the stimulus of an NLRP3 inflammasome, and wide-field imaging high content is used for capturing fluorescent cells. The fluorescence sensor can achieve two goals, first, substances causing high expression of NLRP3 can be discovered rapidly and efficiently, and second, in the role of an NLRP3 activator, substances inhibiting NLRP3 activation can be quickly and efficiently found. The wide-field imaging high content can be used for shoot fluorescent images in a high-throughput mode, and the efficiency of screening medicines is greatly improved. The method disclosed by the invention can be used for primarily screening of drugs with the NLRP3 inflammasome as a target, and can also be used for the scientific study of the NLRP3 inflammasome.

A process of in vivo labeling and identifying recombinantly produced peptides or proteins within an unpermeabilized prokaryotic host cell. Recombinant prokaryotic cells expressing a fusion peptide comprising at least one tetracysteine tag were labeled in vivo using a biarsenical labeling reagent. A fluorescent activated cell sorter was used to identify and select subpopulations of fluorescent cells wherein the amount of fusion peptide in the cell was proportional to the amount of fluorescence detected.

The invention discloses a cell fluorescence image discrimination method and system. The cell fluorescence image discrimination system comprises a format conversion module, an image binarization module, a fluorescence pattern position collection module, a cell fluorescence image cutting module, a cell classification module, a data set classification module, a sample equalization module, a data preprocessing module, a mathematical model construction module and a mathematical model training module. And a mathematical model test module and a detection module. According to the cell fluorescence image discrimination method and system provided by the invention, the efficiency and accuracy of fluorescence cell recognition can be improved. In a use scene of the invention, cell segmentation is carried out by using a fluorescence image in an HSV format, then positive and negative detection is carried out on segmented cells by using a deep learning network model, and finally positive cell quantity statistics is carried out, so that classification and recognition of the fluorescence staining image are realized.