360 results about "Fluorescent staining" patented technology

An improved method of making a series of bead or microsphere or particle populations characterized by subtle variation in a proportion or ratio of at least two fluorescent dyes distributed within a single bead of each population is provided. These beads, when excited by a single excitation light source are capable of giving off several fluorescent signals simultaneously. A set containing as many as 64 distinct populations of multicolored, fluorescent beads is provided and when combined with analytical reagents bound to the surface of such beads is extremely useful for multiplexed analysis of a plurality of analytes in a single sample. Thus, methods of staining polymeric particles, the particles themselves, and methods of using such particles are claimed.

The invention relates to an amphiphilic illuminant with aggregation induced emission characteristics and applications thereof. The illuminant is prepared by connecting a hydrophilia unit on a classical hydrophobicity unit of the classical aggregation induced emission characteristics (AIE), and can be applied to a fluorescent light chemical sensor and is used for preparing fluorescent light coloring agent which is used for coloring living cells and animal imaging fluorescent light. The amphiphilic coloring agent is specifically suitable for the fluorescent light mark on biopolymer, and can be used as a biocompatibility probe for AIE activation so that the amphiphilic coloring agent can be applied to clinic cancer imaging, diagnose and treatment.

A method of detecting and quantifying viable cells in a sample. The method includes fluorescently staining the cells by adding a fluorescent dye into the sample or putting the sample in contact with the fluorescent dye. A quenching dye is then added to the stained sample, or the sample is put into contact with the quenching dye, at a pH different from the pH in the viable cells. The quenching dye used is permeable through the membrane of a viable cell and does not readily absorb fluorescence of the fluorescent dye at the pH in the viable cells, but absorbs the fluorescence of the fluorescent dye at the pH of the fluorescent dye. Next, the sample, now stained with the fluorescent dye and the quenching dye, is illuminated by an excitation light for the fluorescent dye at a pH different from the pH in the viable cells and the fluorescence emitted from the sample is collected and detected.

The invention belongs to the field of high polymer materials, and discloses a composite fiber containing aggregation-induced luminescent molecules, a preparation method thereof and an application thereof. The preparation method of the composite fiber comprises the steps of adding aggregation-induced luminescent molecules into a high polymer material solution or a solution of a high polymer material and a carrier, uniformly mixing the solution, and preparing the composite fiber through the electrostatic spinning process. The composite fiber containing aggregation-induced luminescent molecules has multiple types of AIE, and is strong in fluorescence intensity. Therefore, the composite fiber can meet the requirements of different fluorescent staining operations. The preparation method provided by the invention is simple, high in production efficiency, and good in repeatability. The composite fiber containing aggregation-induced luminescent molecules can be applied in the fields of cell positioning and imaging, fiber microstructure observation, fiber drug loading and the like.

The invention relates to a circulating tumor cell detection kit and application thereof. The circulating tumor cell detection kit comprises a chip, sample diluent, a cell separation fluid, a cell trapping agent, a cell cleaning solution, a cell fixing agent, antibody diluent, a cell penetration agent and a fluorescence dye. The application method of the circulating tumor cell detection kit mainly comprises the several steps of separation of circulating tumor cells, trapped antibody coating, trapping of the circulating tumor cells and immuno-fluorescent staining of the circulating tumor cells. The circulating tumor cell detection kit is mainly used for tumor prognosis, reappear, metastasis, curative effect monitoring, early diagnosis and early warning, monitoring of curative effect and tumor progression situation, and utilizes the molecular subtyping, screening specificity and other characteristics of the circulating tumor cells to conduct targeted research on circulating tumor cell sensitive drug so as to supplement tumor metastasis, relapse and drug resistance mechanisms and guide individualized medical treatment.

The invention relates to an immunofluorescence staining method for suspension cells. The immunofluorescence staining method for the suspension cells is characterized by comprising the following steps of: collecting cells into a centrifuge tube; performing 800-gram centrifugation to collect the cells; respectively adding paraformaldehyde to fix, Triton-X100 to pass through and 1 percent bovine serum albumin (BSA) to close; adding a primary antibody and a secondary antibody sequentially to incubate; washing by an 800-gram centrifugation method after the operation of each step; performing diamino phenyl indole (DAPI) staining; and dripping on a glass slide and closing the glass slide to observe. The immunofluorescence staining method for the suspension cells has the advantages that: the immunofluorescence staining process is performed in the centrifuge tube, so the problems that the suspension cells grow on the glass slide difficultly and drop off easily in the test process are solved; the staining result is good; and the method is suitable for the suspension cells and cells which are adhered to the wall difficultly.

Systems and methods for analyzing blood samples, and more specifically for performing a white blood cell (WBC) differential analysis. The systems and methods screen WBCs by means of fluorescence staining and a fluorescence triggering strategy. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder WBC reagent(s), suitable for assays of samples containing fragile WBCs. In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye, which corresponds in emission spectrum to an excitation source of a hematology instrument; (b) using a fluorescence trigger to screen the blood sample for WB Cs; and (c) using measurements of (1) axial light loss, (2) intermediate angle scatter, (3) 90° polarized side scatter, (4) 90° depolarized side scatter, and (5) fluorescence emission to perform a differentiation analysis.

The invention relates to a method for fluorescent staining of orlon, which belongs to the field of dye synthesis and textile dyeing. Stilbazole salt is taken as a cation fluorescent dye, 0.2 and 2.5 percent owf fluorescent dye, 0.5g / L Peregal 0, a 1.0 percent owf leveling agent and 4g / L glauber salt are mixed to prepare a dye solution, acetic acid and a sodium acetate buffer solution are used to adjust the pH value of the dye solution to 5, and the dyeing processing with the dyeing bath ratio of 100 to 1 is performed to obtain a fluorescent orlon fabric. The stilbazole salt provided by the method has simple synthesis reaction, has relative flexibility in molecular design and assembly, and also has good linear optical performance and non-linear optical performance; and under specific dye bath concentration, the dyed orlon fabric satisfies the coordinate range of orange color stipulated in European EN471 standard High-visibility Warning Clothing for Professional Use-Test Methods (2003).

A fluorescent paint used for reinforcing infrared light comprises an infrared long afterglow luminescent material, diluent, binding agent and a fluorescence staining substance, and is characterized in that the infrared long afterglow luminescent material is mixed with the diluent to form a fluorescent solution containing the infrared long afterglow luminescent material, the fluorescent solution and the binding agent are evenly mixed to form fluorescent binding agent, and the fluorescent binding agent is combined with the fluorescence staining substance. The fluorescent paint used for reinforcing infrared light absorbs ultraviolet light in sunlight and releases near-infrared light for a long time at night. The near-infrared light can be sensitively detected by an existing CCD camera. The fluorescent paint used for reinforcing infrared light can form a static infrared light-emitting environment background when painted on fencings, walls, cloth, trees and the ground. The fluorescent paint used for reinforcing infrared light has the advantages of being stable, free of toxin, environmentally friendly, low in cost, simple to use and the like, thereby being capable of being widely applied to the fields of security monitoring, night target infrared displaying, dynamic tracking without heat source, anti-fake fluorescent materials and the like.

The invention belongs to the field of high-polymer materials and discloses a high-polymer vesicle containing AIE (aggregation-induced emission) molecules as well as a preparation method and an application of the high-polymer vesicle. The high-polymer vesicle is mainly formed through self-assembly of the AIE molecules and amphiphilic block copolymers; the outer layer and the inner layer of a high-polymer vesicle film are hydrophillic layers, each hydrophillic layer is formed by hydrophilic chain sections in the amphiphilic block copolymers, and a film intermediate layer between the outer layer and the inner layer is formed by hydrophobic chain sections in the amphiphilic block copolymers and the AIE molecules. The high-polymer vesicle has high fluorescence intensity and can meet different fluorescent staining demands. The preparation method is simple and feasible, the production efficiency is high, and the repeatability is good. The high-polymer vesicle can be used for related fields of optical bioimaging, biological detection and clinical imaging, high-polymer vesicle research and the like.

The present invention discloses a simple single circulating tumor cell separation method and apparatus. In particular, the present invention discloses a single circulating tumor cell (CTC) separation method, first, an enriched CTC sample is obtained by negative enrichment of a peripheral blood sample; a first-fluorescent-labeled anti-epithelial cell marker antibody and a second-fluorescent-labeled anti-CD45 antibody are respectively used for fluorescent staining of the enriched CTC sample, and the stained enriched CTC sample is diluted; and single cell isolation is performed on first fluorescent positive and second fluorescent negative target cells. The present invention also discloses a single circulating tumor cell (CTC) separation apparatus applicable to the single circulating tumor cell (CTC) separation method. CTC negative enrichment technology is used for the CTC enrichment, compared with a positive magnetic bead enrichment method, disturbance to the target cells is less, and the picked and separated cells can completely maintain cell activity, can be used for nucleic acid and protein analysis, and can also be used in cell culture.

A method for distinguishing erythrocytes in a biological specimen, comprising the steps of preparing a sample liquid by performing to give a damage to a cell membrane of yeast-like fungi without hemolyzing erythrocytes in a biological specimen and to stain the yeast-like fungi with a fluorescent dye; detecting a first information and a second information from a particle in the sample liquid, wherein the first information reflects a size of the particle and the second information reflects a degree of fluorescent staining of the particle; and distinguishing the erythrocytes from the yeast-like fungi based on the first information and second information detected, is disclosed. An apparatus, a reagent kit and a reagent for carrying out the method are also disclosed.

Systems and methods for analyzing blood samples, and more specifically for performing a nucleated red blood cell (nRBC) analysis. The systems and methods screen a blood sample by means of fluorescence staining and a fluorescence triggering strategy, to identify nuclei-containing particles within the blood sample. As such, interference from unlysed red blood cells (RBCs) and fragments of lysed RBCs is substantially eliminated. The systems and methods also enable development of relatively milder reagent(s), suitable for assays of samples containing fragile white blood cells (WBCs). In one embodiment, the systems and methods include: (a) staining a blood sample with an exclusive cell membrane permeable fluorescent dye; (b) using a fluorescence trigger to screen the blood sample for nuclei-containing particles; and (c) using measurements of light scatter and fluorescence emission to distinguish nRBCs from WBCs.

The invention discloses application of a ruthenium complex serving as a nucleic acid vector of a target cell nucleus. According to experimental data, the ruthenium (II) complex can be effectively combined with nucleic acid sequences and effectively change forms of long-sequence nucleic acids to enable the nucleic acids to be effectively transported into living cells in a transmembrane manner and well positioned in cell nucleuses, so that nucleic acid transporting efficiency is greatly improved. Therefore, the nucleic acid sequences can be conveniently transferred into the cells to realize gene therapy, fluorescent staining tracing or the like. By a ruthenium complex-nucleic acid compound preparation method, stable ruthenium complex-nucleic acid compounds can be prepared efficiently, and better effects are achieved.

The invention relates to a fluorescent dyeing method for observing the microstructure of plants, which can effectively solve the problem of rapid film production and observation of the microstructure of plants. , the plant material was fixed in paraformaldehyde or methanol with a mass concentration of 4% configured by pH 8.0 PBS buffer solution for 20-40min, rinsed with pH 8.0 PBS buffer solution for 10-20min, and prepared by DMSO, Triton X-100 and fluorescent whitening agent 220 make staining solution, stain at room temperature for 10-60min, then rinse with PBS buffer for 10-15min, add DAPI solution dropwise for fluorescent staining of nucleic acid in cells, rinse with distilled water, and then add dropwise Glycerin, add a cover glass, observe and take pictures with a laser confocal microscope, the method of the present invention is simple, does not need a microtome, and the production time is short, and the three-dimensional structure of the plant can be observed, which is suitable for the observation of cells or tissues of various plants, clear , accurate, low cost and good effect.

A method of detecting and quantifying viable cells in a sample. The method includes fluorescently staining the cells by adding a fluorescent dye into the sample or putting the sample in contact with the fluorescent dye. A quenching dye is then added to the stained sample, or the sample is put into contact with the quenching dye, at a pH different from the pH in the viable cells. The quenching dye used is permeable through the membrane of a viable cell and does not readily absorb fluorescence of the fluorescent dye at the pH in the viable cells, but absorbs the fluorescence of the fluorescent dye at the pH of the fluorescent dye. Next, the sample, now stained with the fluorescent dye and the quenching dye, is illuminated by an excitation light for the fluorescent dye at a pH different from the pH in the viable cells and the fluorescence emitted from the sample is collected and detected.

The invention belongs to the technical field of biotechnology and relates to the sequences of five oligonucleotide aptamers for specifically identifying staphylococcus aurei and use thereof. The fivesinge-chain DNA oligonucleotide aptamers for specifically identifying the staphylococcus aureus are obtained by a bacterial subtraction SELEX technology. After FITC is marked, the five aptamers are proved by fluorescent staining to be capable of specifically identifying the strain of the staphylococcus aurei. After being used in combination, the five ligands can improve sensibility in identification of target bacteria and the ratios of detected different staphylococcus aureus strains and the detected staphylococcus aurei in different growth states. Therefore, the sequences of the five aptamershave wide application prospects in quick detection of the staphylococcus aurei in samples including suspected terrorist attack biological agents and identification of the staphylococcus aurei and other gram-positive cocci.

The invention relates to a fluorescent staining kit for rapid detection on biological cell viability, and application of the same. The kit comprises an anthocyanin dye and a propidium iodide dye. The detection comprises the following steps of: adding the anthocyanin dye and the propidium iodide in the kit are added in cell suspension, observing and counting the cells by a fluorescence microscope or detecting the cells by a microporous fluorescent plate reader under wavelength of emitted light, and reflecting the viability of the cells in biologic sample liquid according to the strength of fluorescence values of different colors. The kit can be used for rapidly observing or detecting the amount of living cells and the amount of dead cells of various cells in the sample in real time, is visual and handy, eliminates the inference of other substances and the defect of long culture cycle, and is not only suitable for animal cells, but also suitable for most gram negative bacteria and gram positive bacteria.

The invention relates to a building method of a zebra fish hyperlipidemia model and application of the animal model to hyperlipidemia disease research and lipid-lowering drug screening. The building method of the zebra fish hyperlipidemia model mainly includes the steps of zebra fish selection, feeding of zebra fish by yolk powder, histochemical staining or fluorescent staining, image analysis and / or microwell plate analysis and statistical analysis. The building method has the advantages of simplicity, convenience, rapidity, economy, high efficiency, high throughput and the like, and the model can be used for hyperlipidemia disease research and lipid-lowering drug screening. The building method and application of the zebra fish bacterial infection model are of great significance to acceleration of research and development on lipid-lowering drugs and improvement on treatment of patients suffering from hyperlipidemia.

The invention provides a micro-fluidic chip for cell capture and fluorescent staining. The micro-fluidic chip comprises a substrate and a cell runner, wherein the cell runner is arranged on the substrate; a sample inlet and a waste liquid outlet are formed in the substrate; one end of the cell runner is a cell inlet; the other end of the cell runner is a cell outlet; the cell inlet is connected with the sample inlet; the cell outlet is connected with the waste liquid outlet; a staining liquid channel is further connected with the cell inlet; a staining liquid inlet is formed in the other end of the staining liquid channel; the cell runner consists of a plurality of micro channels which are arranged in parallel from head to end; a cell capturing device is arranged inside the micro channels. The micro-fluidic chip provided by the invention is high in capturing efficiency and small in cell injury, the integration of enrichment, capturing and staining of target cells can be achieved, artificial interference can be reduced, and the sensitivity and the reliability of results can be improved. The micro-fluidic chip provided by the invention is free of complex process, can be prepared by using a common method, and is low in cost and very good in practicability.

The invention provides a cell interpretation method and system based on an FISH technology. The cell interpretation method comprises steps of acquiring a to-be-interpreted cell image; acquiring the morphological information and the signal intensity information of fluorescence staining signal points in the to-be-interpreted cell image; determining the type information of each fluorescence stainingsignal point based on the morphological information and the signal intensity information; wherein the type information comprises any one of a normal signal point, a strong signal point, a weak signalpoint, a split signal point and a rod-shaped signal point; based on the type information of each fluorescent staining signal point, determining the quantity information of the fluorescent staining signal points of each staining channel in the to-be-interpreted cell image; and judging whether cells in the to-be-interpreted cell image are circulating abnormal cells or not based on the quantity information of the fluorescent staining signal points of each staining channel. The method is advantaged in that technical problems that in the prior art, in the automatic analysis and treatment process ofthe FISH stained cells, accuracy is low, and the flux in the manual interpretation process is low are solved.

An immune fluorescent microscopic observing method of cytula micro-pipe skeleton from bothidae flasfish includes applying fixture solution to fix sample and utilizing dissection needle to separate germ disc from embryon, making hole on sample and using sodium borohydride to eliminate aldehyde group influence on experimental result, carrying out indirect immune fluorescent staining on sample then using fluorescence microscope to observe sample.

Disclosed are activity evaluation of plant pathogens and a high throughput screening method for microbicides based on double fluorescent staining and a kit therefor, which include using a microtiter plate as tool carrier, using two different fluorescent dyes, respectively fluorescently labelling spores or mycelia of plant pathogens having vitality and no vitality, and due to the different colouring of spores or mycelia having no vitality and vitality, quantitatively detecting the survival rate of plant pathogens through fluorescence microscopy or flow cytometry, and accordingly preparing the kit. The method and kit can be used in aspects such as microbicide screening, evaluation of food safety, and water quality evaluation using the plant pathogen survival rate as an indicator. The method and kit are rapid, objective, simple and easy in operation, and have a low cost, and both can be used in studying the mechanism of the action of microbicides and high-throughput screening and the toxicity evaluation of microbicides.

The invention discloses a solvent-extraction-and-fluorescence determination method of microalgae lipid content, and belongs to the technical field of microalgae crude lipid determination. The solvent-extraction-and-fluorescence determination method of the microalgae lipid content solves the problems of a fluorimetry in the prior art that dyeing with direct use of Nile red fluorescent staining is halfway and disturbance of microalgae body chlorophyll fluorescence peaks and the like occur when an ultrasound method or a dimethyl sulfoxide (DMSO)-processing-Nile-red-dyeing fluorescence determination method is in use. The solvent-extraction-and-fluorescence determination method of microalgae lipid content comprises the following steps of preparing a standard curve by using microalgae to prepare standard microalgae liquid, extracting microalgae lipid and determining content of the microalgae lipid by using fluorescence. Due to the fact that in a determination process, operation steps are free from evaporation organics, and pollution to the environment is little. The solvent-extraction-and-fluorescence determination method of the microalgae lipid content has the advantages that the solvent-extraction-and-fluorescence determination method of the microalgae lipid content is rapid, simple, effective and sensitive, and determination results are free from influence of retention periods of microalgae samples.

The present invention discloses a super-thick tissue slice preparation method, which comprises: obtaining a tissue material to be observed; preparing tissue material slices with a thickness of 1-8 mm,and carrying out transparentizing on the tissue material slices; and carrying out fluorescent staining on the transparent tissue material slices. The method for reproducing the three-dimensional tissue structure of a tissue slice comprises: preparing a transparent tissue slice with a thickness of 1-8 mm; obtaining the image data of the transparent tissue slice by using a fluorescence microscope;recombining the image data to form three-dimensional model data; and based on the three-dimensional model data, outputting a three-dimensional image. According to the present invention, with the super-thick tissue slice preparation method and the three-dimensional tissue structure reproducing method, the tissue slice is subjected to grease removing to achieve the transparent state so as to increase the penetrating depth of laser in liver tissue and reduce the light scattering, such that the deep imaging of the tissue can be achieved, the real cell structure of the tissue can be reflected, andthe imaging efficiency and the fidelity can be improved.

The invention discloses a floating sphere immunofluorescent staining method and a staining device, belonging to methods for coloring a sample for test. The staining method comprises the following steps: separation of spheres from a culture medium, fixing by stationary liquid, adding of Triton X-100 transparent cells, closing by confining liquid, adding of an interest protein-resisting primary antibody working solution, adding of a primary antibody-resisting fluorescent secondary antibody working solution, and cell nucleus staining by DAPI dye liquor. The staining device comprises a cell chamber and a staining sleeve part arranged at the outer part of the lower end of the cell chamber in a sleeving way. The floating sphere immunofluorescent staining method has the advantages of high efficiency, simpleness, convenience and the like, an antibody usage amount is reduced, experiment cost is reduced, manpower and time are saved, an accurate and reliable experiment result is obtained on the basis that a fundamental principle that immunofluorescent staining is not changed and specially training experimenters is not needed, sphere loss in an experiment operation process is avoided, an experiment success rate is increased, and the efficiency is improved.

The invention discloses an integrated drug screening and staining micro-fluidic chip, which is such structured that a liquid path control layer is arranged on the upper layer of the micro-fluidic chip, a gas path control layer is arranged on the lower layer, and a blank glass base plate is arranged on the bottom surface of the micro-fluidic chip; and specifically, the micro-fluidic chip structurally comprises a cell fluorescent staining sample inlet, a fluorescent staining sampling channel area, a cell sample inlet, a cell sampling channel area, a cell culture room, a drug sampling channel area, a drug sample inlet, a liquid outflow channel area and a liquid outlet. The invention also provides a preparation method of the integrated drug screening and staining micro-fluidic chip, wherein the preparation method of the integrated drug screening and staining micro-fluidic chip comprises the following steps: preparing a photoresist template that channel parts protrude; promoting developingand hardening; processing the template by virtue of a silylating reagent; obtaining a polydimethylsiloxane chip having a structure; and implementing irreversible sealing. The micro-fluidic chip provided by the invention is simple in structure, convenient in preparing operations, high in speed, high in efficiency and broad in application scope.

The invention discloses a fluorescent staining method for a cotton fiber. The method comprises the specific steps of pretreatment, modification, staining and fixation, wherein in the pretreatment step, a cloth sample is pretreated; in the modification step, water is added and then a modifier is added to the pretreated cloth sample to ensure that original negative charge on the surface of the cotton fiber is changed to positive charge, operation is conducted for 5min, temperature is raised to 70 DEG C at 1.5 DEG C per minute and kept for 30min, the cloth sample is taken out of a cylinder and washed with water once, and the modification is finished; in the staining step, water is added and then fluorescent paint for printing is added according to a color formula to the modified cloth sample, operation is conducted for 5min, temperature is raised to 70 DEG C at 1.5 DEG C per minute and kept for 30min, the cloth sample is taken out of the cylinder and washed with the water once, and the staining is finished; and in the fixation step, water is added and then an adhesive is added to the stained cloth sample, temperature rise and heat preservation are conducted, and the cloth sample is taken out of the cylinder, washed with the water cleanly, softened and dried. According to the fluorescent staining method for the cotton fiber, the fluorescent paint for the printing can be used for staining, and the hand feeling, fastness and cloth cover uniformity are improved.

A method of utilizing nuclear specific fluorescent staining and ovary being made to be completely transparent technique to observe blastocyst of rice includes using certain concentration of sodium hydroxide solution to carry out softening treatment on blastocyst of rice before staining then utilizing specific combination of nucleus fluorescent staining DAPI with nucleus to observe structure of cell and nucleus in blastocyst, enabling to use laser scan confocal microscope to observe internal structure of cell clearly.