349 results about "Fluorescein isothiocyanate" patented technology

The invention belongs to the technical field of nano materials and biomedicine, in particular to multifunctional core-shell structure fluorescent coding magnetic microspheres and a preparation method thereof. On the basis of ferroferric oxide nanoparticles synthesized by a hydrothermal process, pre-prepared coupled product of fluoresceins and amino propyl trimethoxysilane and ethyl orthosilicate are subjected to cohydrolysis in ammonia water to form a multifunctional fluorescent magnetic nano composite material with fluorescent and magnetic properties and high biological stability and biological adaptability. By regulating the mixing ratio of fluoresceins, namely fluorescein isothiocyanate (FITC) and rhodamine B isothiocyanate (RBITC), various fluorescent coding magnetic microspheres can be prepared, and the particle size of obtained material can be regulated according to the different ratios of the added ethyl orthosilicate to the ferroferric oxide. In addition, the obtained multifunctional composite nano material is subjected to amino silanization modification, so the biological application range of the novel fluorescent coding magnetic microspheres is further expanded and the novel fluorescent coding magnetic microspheres have bright application prospect in fields of biomedicine technology, medicine development, suspension chip and the like.

The invention discloses a preparation method of fluorescent complex microgel which is sensitive to temperature and pH. The method comprises the technical steps of preparing a mixed surface active agent, preparing the oil phase of a template, preparing an emulsive liquid, preparing a water phase, preparing N-isopropyl acryl amide copolymerized methacrylic acid microgel, preparing turgid N-isopropyl acryl amide copolymerized methacrylic acid microgel, preparing a mixed liquid of Gamma-aminopropyl triethoxy silane and normal heptane, preparing a silicon dioxide polymer complex microballoon decorated by a surface deposited amino group, preparing a mixed liquid of fluorescein isothiocyanate and absolute ethyl alcohol as well as preparing the fluorescent complex microgel. In the invention, the fluorescein isothiocyanate with high quantum yield and good optical stability is adopted as fluorescent matter; the prepared fluorescent complex microgel is sensitive to temperature and pH; and the size of the prepared fluorescent complex microgel is in a micron range and is hard to be aggregated. The method has the advantages of reasonable design, practical technique and easy operation; moreover, reactions are carried out under normal temperature and the like; the method can be applied to the fields of controlled release of medicament, biological probe, chemical separation, and the like.

The invention provides an aflatoxin B1 (AFB1) magnetic separation enzyme-linked immunity quantitative detection method, belonging to the field of food safety immunoassay technique. The method adopts the immuno-detection principle of competition law; and AFB1 is connected with biological enzyme to prepare enzyme-labeled antigen reagent, anti-fluorescein isothiocyanate (FITC) antibody is absorbed onthe surfaces of magnetic particles to prepare magnetic separation reagent, and the FITC is connected with the AFB1 antibody to prepare anti-reagent. In a sample, the AFB1 competes with the enzyme-labeled AFB1 and is combined with a small amount of FITC-labeled anti-AFB1 antibody, so that antigen-antibody complex can be formed. After the magnetic separation reagent is added, the complex is caughtonto the surfaces of the magnetic particles by the anti-FITC antibody connected on the surfaces of the magnetic particles. After being washed, the product is finally added with substrate and detected.The method has the advantages that (1) the magnetic particles are used for replacing the traditional enzyme-labeled plate to be taken as a solid-phase carrier, so that immunoreaction is carried out under the approximate liquid phase condition; and the reaction is more complete and rapid, and has the characteristics of high specificity and good repeatability compared with the traditional enzyme-linked immuno sorbent assay (ELISA); furthermore, (2) by adopting one-step competition law principle, the time used for detection is short.

The invention discloses primers for loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) detection of a vibrio vulnificus and probe sequences. The primers are characterized by including three pairs of LAMP primers, namely TolC-F3, TolC-B3, TolC-FIP, TolC-BIP, TolC-LF and TolC-LB, and a probe TolC--HP, wherein the Van-FIP is a biotin labeling primer at 5' terminal; the probe TolC-HP is a fluorescein isothiocyanate FITC labeled probe at the 5' terminal; concrete sequences are shown in SEQ NO1-NO7 in a sequence table. The primer has the advantages of good quickness, specificity and sensitivity and simple demands on instruments; early diagnosis and detection of the vibrio vulnificus are facilitated; the demands of a basic-level detection mechanism and field epidemic focus detection can be met.

The invention discloses a quantum dot-organic dye compound closed/open type cadmium ion ratio fluorescent probe and a preparation method thereof. The preparation method comprises selecting CdTeQDs to serve as a main fluorescence source, introducing fluorescein isothiocyanate (FITC) to serve as an auxiliary (or reference) chromophore, and forming a CdTe-FITC compound dual-emitting probe in a hydrogen bonding mode. The problem that fluorescence signals of a traditional single-fluorescence probe are difficult to obtain accurately is solved. A fluorescence closed/open model is adopted, disturbance of factors except an analyte on a fluorescence enhancement reaction can be effectively reduced, the fluorescence enhancement reaction has specificity, and accordingly sensitivity and selectivity of analyte detection are improved. The preparation method is simple and low in cost, and the prepared product can perform accurate, sensitive, simple, convenient and rapid detection on Cd2+ in complex systems and biological samples, has important reference values for developing high-quality fluorescence probes of other types and provides a new developing direction for research in the fields of biological detection and chemical analysis and the like.

The invention discloses a constant-temperature amplification method of double-labeled nucleic acid and a test strip. The constant-temperature amplification method comprises the following steps of: adding labeled primers into an LAMP (loop-mediated isothermal amplification) reaction system for constant-temperature amplification by firstly synthesizing inner primers, outer primers and ring primers and using biotin and fluorescein to label the inner primers or/and the ring primers, and obtaining a constant-temperature amplification product of the double-labeled nucleic acid. The test strip comprises a supporting layer and a nucleic acid amplification product adsorbing layer which is pasted on the supporting layer; and the biotin affinity protein and fluorescein isothiocyanate antibody are used for printing a detection mark and a control mark on a cellulose layer. In the test strip disclosed by the invention, the detection specificity is strong, the sensitivity is high, the speed is high,not only can the LAMP product be intuitively detected, but also the defect that the specificity of the detection test paper of the colloidal gold is lower can be overcome. The invention can be widelyused for detection of specific genes in various pathogenic microorganisms or pathogens.

The invention discloses a kit using a magnetic particle chemiluminiscence method to measure osteocalcin (BGP) content. The kit comprises a calibration article, a quality control article, a resisting reagent, a magnetic particle reagent and a light-emitting substrate, wherein the resisting reagent comprises an antibody marked by fluorescein isothiocyanate and coated by osteocalcin and an antibody marked by alkaline phosphatase and osteocalcin; the magnetic particle reagent is used for connecting magnetic particles with anti-goat FITC. The kit has the advantages that the chemiluminiscence technology is combined with immune magnetic particles, a reaction system close to homogeneous phase is provided, a one-step reaction mode is used, the detection sensitivity and precision of the kit are increased greatly, the detection range of the kit is increased, reaction time is shortened greatly, time from sample feeding to a detecting result is less than 35 minutes, and the detection speed of the kit is evidently higher than that of similar kits; the kit can detect multiple samples at the same time on full-automatic chemiluminescence apparatus, high-throughput and fast measuring of the osteocalcin is achieved, the kit is high in accuracy and high in specificity, and the accuracy and detection efficiency of the kit are increased greatly.

The invention relates to a silica fluorescent nanoparticle and a preparation method thereof, in particular to an FITC-doped silica fluorescent nanoparticle and the preparation method thereof. The method particularly comprises the preparation of a precursor solution and other steps. The FITC is reacted with 3-aminopropyl trimethoxysilane to prepare a precursor, and then the novel FITC-doped silica fluorescent nanoparticle is prepared by a water-in-oil microemulsion method. The prepared nanoparticle is uniformly spherical, has good monodispersity and light stability, difficultly generates dye leakage, has good response to the pH value and can be applied to the real-time monitoring of the pH value of cells.

The invention discloses a C-reactive protein quantitative determination kit, belonging to the field of biological immune in-vitro diagnosis of medical devices. The kit comprises a calibrator, a quality control product, an antibody reagent, a magnetic particle reagent and a luminescent substrate. The invention also discloses a preparation method of the kit and a method for detecting C-reactive protein by using the kit. According to the preparation method disclosed by the invention, the antibody reagent is prepared from a C-reactive protein-coated antibody labeled by fluorescein isothiocyanate and a C-reactive protein-labeled antibody labeled by alkaline phosphatase, and the magnetic particle reagent is prepared by coupling an anti-fluorescein-isothiocyanate antibody with carboxyl magnetic beads so as to ensure that uniform mixing and separation are easily carried out in immunoreaction, and the reaction speed can be greatly increased; and a novel chemical luminescent substrate ALPS is used as the substrate, so that the sensitivity and specific performances of the kit can be improved. The detection kit disclosed by the invention is reliable in performance, high in sensitivity and wide in linear range, and can be matched with semi-automatic and full-automatic instruments for use.

The invention relates to a magnetic particle chemiluminescence immunoassay method for human thyroglobulin antibodies (TGAb) and belongs to the technical field of immunoassay. TG antigens marked by fluorescein isothiocyanate (FITC) and human IgG antibodies marked by alkaline phosphatase are combined to form an antigen-antibody-second enzyme-labeled antibody sandwich immune complex with a sandwich-like structure. Then, magnetic particles connected with FITC antibodies are added, the antigen-antibody complex is connected to the magnetic particles through specific combination of the FITC antibodies and the FITC and directly deposited in an external magnetic field, and the complex formed by immune reaction can be separated from the other non-combined substances without centrifuging. A kit combines chemiluminescence with the magnetic particles to provide a reaction system close to a homogeneous phase. Compared with the prior art, the kit has the advantages of higher sensitivity, wide linear range, quickness and the like; the cost of a product is greatly reduced; and the kit has a broad application prospect on the aspects of clinical examination and the like.

The invention discloses a kit for detecting procalcitonin. The kit comprises magnetic particles enveloped by a polyclonal antibody for resisting fluorescein isothiocyanate, a procalcitonin monoclonal antibody labeled by fluorescein isothiocyanate, a procalcitonin monoclonal antibody labeled by alkaline phosphatase, and chemiluminescent substrate liquid which is catalyzed by alkaline phosphatase to emit light. The kit for detecting procalcitonin, which is disclosed by the invention, adopts a double-antibody sandwich reaction mode and effectively makes use of a theory of combining a chemiluminescent detection technology and a magnetic particle immune separation technology to quantitatively measure the PCT (procalcitonin) content of a serum or plasma sample of a human body, so that the sensitivity of detection is guaranteed; furthermore, all indexes can reach the analysis level of the same type of imported kits.

The invention relates to a method for preparing a magneto-optic difunctional CNT/Fe304@SiO2(FITC) (Fluorescein Isothiocyanate) one-dimensional nano composite material. The method comprises the following steps of: 1. preparing CNT/Fe304; 2. preparing APS (3-Aminopropyl-Trimethoxysilane)-FITC; and 3. preparing the CNT/Fe304@SiO2(FITC) composite material. The prepared magneto-optic difunctional CNT/Fe304@SiO2(FITC) one-dimensional nano composite material is applied to the aspects of targeted visible medicine carrying, preoperative diagnosis, intraoperative treatment and photoelectric device assembling.

The invention discloses a magnetic particle chemiluminescence immunoassay kit of free thyroxine (FT4), belonging to the medical field of immunoassay. The kit comprises: 1) free thyroxine series calibrator; 2) magnetic particle solution enveloped by anti-fluorescein isothiocyanate monoclonal antibody; 3) free thyroxine antibody solution marked by fluorescein isothiocyanate; 4) free thyroxine markedby horse radish peroxidase; 5) chemiluminescence substrate liquid; and 6) concentrated washing liquid. The invention also discloses a preparation method of the kit. The kit in the invention has advantages of simpleness, convenience, rapidness, sensitivity, stability and the like.

The invention provides a clenbuterol (CLB) magnetic particle separation enzyme-linked immunosorbent assay (ELISA) method, belonging to the technical field of food safety immunodetection. The method comprises the following steps: the immunodetection principle of the competition method is adopted to connect CLB with bio-enzyme and prepare an enzyme labeled antigen reagent, antibody against fluorescein isothiocyanate (FITC) is absorbed on the surface of magnetic particles to prepare a magnetic separation reagent, FITC is connected with the antibody against CLB to prepare an antibody reagent; CLB in a sample and enzyme-labeled CLB compete to combine with a small quantity of FITC-labeled antibody against CLB and form an antigen-antibody composite; after the magnetic separation reagent is added, the antibody against FITC connected with the surface of magnetic particles captures the composite on the surface of magnetic particles; and washing, and finally adding substrate to detect. The invention has the following advantages: (1) magnetic particles replace the traditional ELISA plate to be used as solid carrier and ensure that the immunoreaction is performed more fully and fast under the near-liquidus condition; compared with the traditional ELISA method, the method is characterized by high specificity and good repeatability; and (2) the one-step competition method principle is adopted, thus the detection time is short.

The invention discloses a magnetic fluorescent multimodal nano biological probe as well as a preparation method and application thereof. The magnetic fluorescent multimodal nano biological probe specifically targets a liver-cancer cell. According to the preparation method, magnetic nano particles and fluorescent dye are co-embedded in silicon dioxide; an amino group is functionally modified by the silicon dioxide; nucleic acid aptamer and a magnetic fluorescent probe are covalently coupled and synchronously specifically target the liver-cancer cell. The magnetic nano particles are paramagnetic iron oxides of which the sizes are ten nanometers; the fluorescent dye is fluorescein isothiocyanate; the embedding medium silicon dioxide is obtained through the hydrolysis of tetraethyl orthosilicate and 3-aminopropyltriethoxysilane; the nucleic acid aptamer is TLS11a. A synthetic method of the probe is simple and convenient; materials are easily obtained; fluorescence properties are stable; a foundation is laid for the early diagnosis of a liver cancer.

Multifunctional “smart” nanostructures are disclosed that include fluorescein isothiocyanate (FITC)-encapsulated SiO₂ core-shell particles with a nanoscale ZnO finishing layer, wherein an outer ZnO layer is formed on the SiO₂-FITC core. These ˜200 nm sized particles showed promise toward cell imaging and cellular uptake studies using the bacterium Escherichia coli and Jurkat cancer cells, respectively. The FITC encapsulated ZnO particles demonstrated excellent selectivity in preferentially killing Jurkat cancer cells with minimal toxicity to normal primary immune cells (18% and 75% viability remaining, respectively, after exposure to 60 μg/mL) and inhibited the growth of both gram-positive and gram-negative bacteria at concentrations ≧250-500 μg/mL (for Staphylococcus aureus and Escherichia coli, respectively). These results indicate that the FITC encapsulated multifunctional particles with nanoscale ZnO surface layer can be used as smart nanostructures for particle tracking, cell imaging, antibacterial treatments and cancer therapy.

The invention relates to a preparation method of a hepatoma carcinoma cell-targeted polyamido-amine dendrimer support, which comprises the following steps of: (1) modifying a polyamido-amine dendrimer by using fluorescein isothiocyanate to obtain G5.NH2-FI; (2) partially acetylizing the G5.NH2-FI to obtain a product of G5-FI-Ac-NH2 in which an amino-group on the surface of the dendrimer is partially acetylized; (3) modifying the G5-FI-Ac-NH2 by using lactobionic acid to obtain G5-FI-Ac-La with functions of targeting a hepatoma carcinoma cell and tracing by fluorescent. The preparation method provided by the invention is simple and has moderate reaction conditions. Furthermore, a used polymer is an environmental-friendly high polymer material and has a prospect of being industrialized. Thefunctional dendrimer material prepared by the invention is good in biocompatibility and can combine a specific target with the hepatoma carcinoma cell, so that the support can be used in load and targeting delivery of medicines, genes or molecule diagnosing probes.

The invention belongs to the technical field of biological medicines and particularly relates to a detection method for anti-CD20 monoclonal antibody binding activities. The method comprises the following steps: a. taking a logarithmic growth phase high-expression CD20 cell, carrying out cell counting, and adjusting the cells to proper cell density for planking; b. carrying out series gradient dilution on a reference and a sample to be detected, and adding culture plates planked with the cells in sequence for a period of time; c. collecting the cells, and adding secondary antibody incubation marked with fluorescein isothiocyanate (FITC) for a period of time; and d. detecting by a flow cytometry to obtain the bonding activities of the sample to be detected. With the adoption of the detection method, the requirements in checking aspects of specificity, precision, linearity, range, accuracy and durability and the like can be satisfied; and the method can be effectively used for detecting the anti-CD20 monoclonal antibody binding activities.

The invention relates to a preparation method of RGD-modified subminiature superparamagnetic iron oxide nanoparticles. The preparation method comprises the following steps of preparing PEI-coated subminiature superparamagnetic iron oxide nanoparticles by a mild reduction method, modifying surfaces of USPIO/PEI.NH2 nanoparticles by fluorescein isothiocyanate (FI) and a RGD compound (COOH-PEG-RGD) subjected to pegylation, and carrying out acetylation treatment on the surfaces of the nanoparticles. The preparation method has simple processes and mild reaction conditions and can be operated easily. The RGD-modified subminiature superparamagnetic iron oxide nanoparticles have small sizes, a high relaxation rate, good colloid stability and biocompatibility, and high alpha<v>beta<3> integrin high-expression tumor cell targeting, and have a latent application value in the field of in-vivo tumor targeting MR imaging diagnosis.

The invention discloses a carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, which comprises a CA 19-9 calibration material, a CA 9-19 quality control material, an anti-reagent, a magnetic micro particle reagent, and a luminescent substrate. The invention also discloses a preparation method of the kit and a method using the kit to detect the carbohydrate antigen 19-9 (CA 19-9). Fluorescein isothiocyanate labeled CA 19-9 coated antibody and alkaline phosphatase labeled CA 19-9 labeled antibody are used to prepare the anti-reagent; anti-fluorescein isothiocyanate antibody is coupled to carboxyl magnetic beads to prepare the magnetic micro particle reagent, thus the materials can be more easily mixed and separated in immunity reactions, moreover, the reaction speed is greatly increased; a novel chemical luminescent substrate APLS is taken as the substrate, and the sensitivity and specificity of the kit are both improved. The provided kit has the advantages of high detection sensitivity, good specificity, and little variation, and the shelf life is one year or more.

The invention provides primers and a method for cross primer isothermal amplification detection of acidovorax citrulli. Nucleotide sequences of the primers areACLF3:5'-GGCTAACTACGTGCCAGC-3'ACLB3:5'-ACGCATTTCACTGCTACA-3'ACLBIP:5'-CAGATGTGAAATCCCCGGGCTCTGCCGTACTCCAGCGAT-3'ACDF5b1:5'-GCAAGCGTTAATCGGAATTACT-3'ACDF5f2:5'-CAACCTGGGAACTGCATTTGT-3', wherein the end 5 of the primer ACDF5b1 is labeled with biotin, and the end 5 of the primer ACDF5f2 is labeled with FITC (Fluorescein Isothiocyanate). The invention further provides a cross primer isothermal amplification technology for detecting the acidovorax citrulli, wherein with total DNA of a sample or a bacterial liquid as a template, isothermal amplification is carried out by using the primers; and after the reaction is finished, a result is judged directly by using a sealed colloidal gold-labeled DNA detecting device. The primers and the method provided by the invention have the advantages of good specificity, high accuracy, high sensitivity as well as simplicity, convenience and rapidness in operation and provides assurance for import/export safety.

The invention belongs to the field of nano composite materials and discloses a preparation method of antibody-mediated optomagnetic double-mode meso-porous silicon nanoparticles. The preparation method comprises the steps of by taking nano ferroferric oxide as a core, applying silicon dioxide to package, labeling by fluorescein isothiocyanate, carrying out surface modification on 3-aminopropyl triethoxysilane to realize amination; then, grafting lactobionic acid by carboxyl-ammonia crosslinking; and finally, activating carboxyl and prostate stem cell antigen (PSCA) antibody. The preparation method disclosed by the invention is simple, and convenient to operate; the obtained nanoparticles have the advantages of active targeting, good biocompatibility, small toxic and side effect, good monodispersity and stability, stable imagnetic and fluorescence performance and the like.

The invention relates to a method for preparing a poly (amidoamine) dendritic polymer/carbon nanometer tube composite material for cancer cell targeting diagnosis. The method comprises that: a carbon nanometer tube CNT is subjected to acidizing treatment; fluorescein isothiocyanate FI and folic acid FA of the poly(amidoamine) dendritic polymer are modified; a CNT composite is obtained through an EDC chemical bonding method and a carboxyl reaction on the surface of a multi-walled carbon nanometer tube which is subjected to the acidizing treatment; terminal amino and acetyl oxide of the rest dendritic polymer on the surface of the composite are subjected to acetylation reaction to obtain a functional composite carbon nanometer tube with neutral surface charge. The functional carbon nanometer tube can be scattered in a solvent for a long time without agglomeration, and the functional multi-walled carbon nanometer tube has good biocompatibility, can be combined on the cancer cell in a targeting manner, and can be used for early-stage target diagnosis of the cancer cell; the method has the advantages of simple preparation method, mild reaction condition and easy operation, and the used polymer is an environment-friendly polymer material, and has industrial implementation prospect.

The invention relates to a method for detecting fungaltoxin by using a photonic crystal microsphere flow type chip. According to the method, a photonic crystal microsphere is used as a fungaltoxin probe carrier; the surface of the microsphere is directly subjected to competitive immune, so that a plurality of fungaltoxins are detected; aflatoxin, Fumonisins and penicillium citrinum toxin are used as detection objects; a fluorescein isothiocyanate marked antibody competes with an antigen fixed on the surface of the microsphere to obtain own antigens; decoding is performed by a confocal fluorescence scanner and a reflection spectrum of a photonic crystal; and the three fungaltoxins can be detected at the same time. According to the method, the detection limit to the aflatoxin, the Fumonisins and the penicillium citrinum toxin is 0.001 ng/mL; the linear detection range is 0.001 to 10 ng/mL; only 10 mu L test sample is needed; and the whole analysis process is shorter than 3 hours.

The invention discloses a magnetic particle Chemiluminescence Immunoassay determination kit of luteinizing hormone (LH), belonging to the field of immunoassay medical science. The kit comprises: 1) LH series of calibrators; (2) magnetic particle solution coated by monoclonal antibody resisting fluorescein isothiocyanate; (3) LH monoclonal antibody labeled by fluorescein isothiocyanate; (4) LH monoclonal antibody labeled by horse radish peroxidase; (5) chemiluminescence substrate liquid; and (6) washing concentrate. The invention also discloses a preparation method of the kit, and the kit has the advantages of simpleness, convenience, rapidness, sensitivity, stability, etc.