423 results about "Microinjection" patented technology

The present invention provides for microinjection devices comprising a needle and a viewing instrument wherein the viewing instrument provides magnified viewing of an object to an operator from an angle other than a right angle to the object.

The invention provides a method for establishing a humanized rat drug evaluation animal model. According to the method, a multidrug resistance gene 1 (Abcb1)-knocked-out genetically engineered rat is obtained through a microinjection method by virtue of a CRISPR/Cas9 gene knockout technology and 153kb bacterial artificial chromosome (BAC) fragments containing a humanized Abcb1 promoter and cDNA is simultaneously inoculated into the rat genome through the microinjection method by virtue of a large fragment transgenic technology to obtain a transgenic rat capable of stably expressing human Abcb1 and the genetically engineered rat and the transgenic rat are hybridized to establish the humanized rat drug evaluation animal model. RT-PCR analysis shows that Abcb1 expression profiles of humanized Abcb1 rat are significantly different from those of the rat endogenous Abcb1. The method has the beneficial effects that the humanized rat capable of expressing human Abcb1 is obtained and the rat is used for expressing human Abcb1 genes and has closer expression profiles to those of human so that the model can be well used for the efficacy evaluation of newly developed drugs.

The present invention provides for microinjection devices comprising a needle and a viewing instrument wherein the viewing instrument provides viewing of an object to an operator from an angle other than a right angle to the object.

An object of the present invention is to provide a method for introducing a physiologically active substance such as a gene into a cell, which introduces a physiologically active substance such as any given gene into any given cell in a view under a microscope, while significantly reducing invasiveness to the cell, and a device used for the above method. The present invention provides a method for introducing a physiologically active substance into a cell, which comprises: allowing a physiologically active substance to attach around a needle having a diameter of 500 nm or less, provided that it is able to be inserted into a cell; and inserting the above-described needle into the cell; and a microinjection device for carrying out the aforementioned method.

The invention discloses a simple, convenient and efficient fixed-point gene editing method relating to the fields of transgenic animal preparation, gene functional study and biomedicines. A testis injection method is a sperm-mediated gene transferring method developed by utilizing the sperm capacity of actively combining, transferring and integrating an exogenous DNA. By using the method disclosed by the invention, an animal testis injection method is optimized, a carrier constructed by utilizing a clustered regularly interspaced short palindromic repeat system (CRISPR/Cas9) is integrated with a sperm chromosome through multi-point testis injection or seminiferous tubule microinjection, so that the aims of deleting, replacing and inserting genes are achieved. Then, a transgenic animal descendant is obtained through various approaches such as natural mating and artificial insemination. According to the method, the advantages of a CRISPR/Cas9 gene editing system are sufficiently utilized, and the existing transgenic animal system is combined, so that the complexity of in-vitro cell operation and requirement on expensive precise instruments/equipment are avoided while the gene modification animal obtaining efficiency is greatly increased.

The invention relates to a preparation method of a shell core fiber covered endovascular stent graft. The steps are as follows: firstly, a polymer is dissolved in adaptive dissolvent to obtain solution with a certain concentration; secondly, medicine or an artificial polymer and medicine, and a bio-active element are dissolved in the adaptive dissolvent to obtain solution or suspension liquid; thirdly, the solution of the polymer and the medicine or the solution or suspension liquid of the medicine or bio-active element are respectively filled in two injectors, the speed of a micro injection pump, the voltage of an electrostatic generator and the distance of a receiving device are adjusted, fiber is obtained through the static spinning preparation, and the fiber is received as a tube shape or film shape structure; fourthly, the endovascular stent graft is fixed on a revolution axle, through the revolution of the endovascular stent graft, the static spinning fiber are directly received as fibrous membrane covered on the endovascular stent graft. The invention can effectively prevent smooth muscle cells from hyperplasia in the stent graft or from narrowing in the stent graft caused by the other functions, and the shell core fiber for the medicine loading can slowly release the medicine to attain the purpose of curing.

The invention relates to a method for acquiring a Nogo-B knockout mode mouse based on CRISPR/Cas9 technology and an application thereof, the method for acquiring the Nogo-B knockout mode mouse comprises the following steps of: designing a high-efficiency sgRNA for identifying EGE-ZXH-007 of a specific gene; connecting the sgRNA to a plasmid vector and then carrying out in-vitro transcription to obtain a Ca9/sgRNA capable of being subject to microinjection; injecting the Ca9/sgRNA capable of being subject to microinjection into a mouse fertilized egg, and then carrying out genotype detection and identification on the born mouse, and finally obtaining the Nogo-B knock-out mode mouse. According to the invention, the Nogo-B knockout mode mouse is obtained by using the CRISPR/Case9 technology,and compared with the traditional gene knockout technology, the operation is easy and the efficiency is higher, and the homozygous mutation is more easily obtained, thus laying a good foundation for further researching the function of Nogo-B.

The invention discloses a method for obtaining sheep with different hair colors on the basis of CRISPR/Cas9 and sgRNA of a targeted ASIP gene. The invention provides sgRNA (ASIP-sgRNA) capable of achieving specific and targeted modification of a sheep ASIP gene, and the sgRNA (ASIP-sgRNA) is RNA as shown from the third nucleotide to the 22<nd> nucleotide of 5' tail end of a sequence 4 of a sequence list or RNA with the nucleotides from the third nucleotide to the 22<nd> nucleotide of the 5' tail end of the sequence 4 of the sequence list. The ASIP-sgRNA particularly can be the RNA as shown in the sequence 4 of the sequence list. The invention further provides a method for obtaining sheep with changed hair colors. The method comprises the following steps that co-transfection of sgRNA capable of achieving specific and targeted modification of the sheep ASIP gene and Cas9mRNA on sheep cells is conducted, and therefore the sheep ASIP gene is deleted, and the sheep with the changed hair colors are obtained. According to the method for obtaining the sheep with different hair colors on the basis of CRISPR/Cas9 and sgRNA of the targeted ASIP gene, a CRISPR/Cas9 genome-editing technology is combined with a microinjection technology, and an effective technological means is provided for artificially changing the hair colors of the sheep.

The invention discloses a unicellular micro-operation device used for microinjection. An air supply and a pressure control valve of the device and a left and a right end effectors form a sealed air course through a passage in a holder; the left and the right end effectors are respectively connected with a left and a right three dimensional micromotion stages through the holders; the left and the right three dimensional micromotion stages are symmetrically arranged on the left side and the right side of an inverted microscope worktable; the left and the right end effectors form a unicellular micro-operation flow field with controllable flow rate distribution on the inverted microscope worktable; and a computer collects signals of an image detecting and processing unit to respectively control the pressure control valve and the left and the right three dimensional micromotion stages. The micro-operation device effectively controls the operation environmental flow field of cells, and forms a stable unicellular micro-operation flow field with controllable flow rate distribution to drive the cells to move. Through the invention, the cellular angle in the three dimensional orthogonalized plane can be randomly adjusted, so that the three degrees of freedom (DOF) attitude of the cells can be accurately controlled; and the three dimensional spatial location of the cells is adjusted to realize the accurate positioning of the location and attitude of the cells.

The invention discloses a method for remarkably improving fish genome editing efficiency. The method for remarkably improving fish genome editing efficiency comprises the following steps: designing a genome editing tool for specifically identifying and cutting a specified site sequence of the fish genome, designing a homologous donor corresponding to the specified site sequence and containing a knock-in exogenous gene fragment, introducing the genome editing tool, the homologous donor and mRNA for specifically and stably expressing fluorescent protein in primordial germ cells into fish animal embryos by using a codominant microinjection method, and selecting the embryos by detecting fluorescent protein expressed by the fluorescent protein mRNA and obtaining stable inheritable characters. The efficiency of the method for knocking the exogenous gene fragment into the specified site of the fish genome so as to obtain a first filial generation of fish with the knock-in exogenous gene fragment is remarkably higher than that of the prior art.

The invention relates to the field of biotechnologies, and provides a construction method for a sialidase gene knockout mouse model and application thereof. An in vitro and vivo targeted mouse genomeNPL is used for genetically recombining a CRISPR knockout plasmid vector, and a diploid double-knockout NPL mouse knockout model is successfully constructed through an oosperm microinjection technology. The NPL has the important function in a pathophysiological process of a body. Vicious transformation of a cancer cell is always along with overexpression of sialic acid. Now, the overexpression ofthe sialic acid has become one of important markers of the malignant transformation and transferring of a tumor. The model is capable of constructing a new research platform for researching a processof cell canceration, beneficial to a key step of researching generation and transferring of the tumor, and providing a foundation for the research and development of a novel anti-tumor drug.

The invention relates to a linear tank-shaped needleless electrostatic spinning device and a spinning method. The device comprises a solution supply system, a spray head system, a solution recycling device and a nanofiber collection system. The solution supply system supplies a solution by controlling an injector through a micro-injection pump; spray heads are linear tank-shaped copper bars; the solution recycling device is a solution tank with the equal length as the copper bar spray heads; the nanofiber collection system is a grounded cylindrical metal roller driven by an adjustable-speed motor. According to the device, the high polymer solution can be automatically and precisely supplied by adopting the micro-injection pump solution supply system; through the linear tank-shaped spray head structure, the problems that when a traditional single needle is adopted, the solution is prone to block, and the nanofiber yield is low are avoided, the purpose of simultaneously forming multiple pieces of jet flow can be achieved, the nanofiber yield in unit time is greatly increased, continuous nanofiber preparation is achieved, and an industrial nanofiber production prospect is achieved.

A cell tray has a multi-dimensional array of cells in precise, equally spaced wells (cubicles or silos) containing medium of interest. The ordered cell array enables automated processing as well as simultaneous monitoring and analyzing of a large matrix of cells, biological fluids, chemicals and/or solid samples. The invention is an integrated device and is fabricated into substrates similar to microscope slides. The ordered array of cells in precise locations helps in parallel analysis and processing of cells simultaneously. Each cell cubicle or silo in the array is located equidistant from its nearest neighbors in an orthogonal direction. The location of each well can be precisely measured and recorded in an automated processing system. Included in the bottom of each cell well is an optional micro-lens. An array of probes provides parallel cell processing and monitoring capabilities, including microinjection and microscope analysis. The cell tray when integrated with the Precision Optical Intracellular Near Field Imaging/Spectroscopy Technology (POINT or NANOPOINT) device results in sub-wavelength high-resolution imaging with a nanosensor array capable of imaging inner regions of living cells without destroying its natural environment.

The invention relates to a preparation of a water-insoluble medicine nanocrystal fibrofelt, which comprises that: water-insoluble medicine is dissolved in organic solvent, then polymeric material is added under the stir condition, the stir is continued and ultrasonic degasification is carried out; the prepared solution is poured into a solution storing tank, a flattened injection needle is used as a capillary tube for injecting thin flow, the positive pole of high voltage source is connected, an aluminum foil receiving flat plate is used to connecte with the negative pole, a microinjection pump is used for controlling the solution injection amount, the spinning condition is adjusted, electrostatic spinning is carried out so that nanocrystal fibrofelt is obtained. The method of the invention is simple, the loading action of polymer material can effectively control the administration in a proper releasing rate and a proper amount, the prepared water-insoluble medicine nanocrystal has favorable solubility, the medicine oral administration bioavailability is improved, multiple ways of administration of water-insoluble medicine is realized, and the application range is widened.

A microinjection apparatus includes a substrate having holes for trapping cells using suction force and injecting a substance into the cells with a needle. Height detection marks are provided on the substrate. A visibility is calculated from an image of a height detection mark, and an amount of deformation of the substrate is determined from the visibility. An XYZ table, on which the substrate is placed, is moved based on the amount of deformation.

A method of expressing in vivo heart-specific fluorescence in transgenic line of zebrafish is developed, which provides a research model for studying heart-related gene functions and performing gene therapies in the future. The method comprises the following steps. Firstly, a plasmid is constructed. This plasmid construct includes the upstream regulatory region, the exon 1, the intron 1, and the exon 2 of cmlc2 gene, cDNA of GFP, wherein the cmlc2 gene and GFP cDNA form a cassette, and inverted terminal repeats from adeno-associated virus are flanked at both sides of this cassette. The plasmid construct is linearized and microinjected into one-celled zebrafish fertilized eggs. Lastly, the heart-specific fluorescent expressed zebrafish are selected and the germline-transmitting transgenic strain is generated.

A container or dish used for the micro manipulation, micro injection, biopsy and fertilization of oocyte and embryo culture. The dish allows the user to more readily perform procedures used to fertilize oocyte, such as intracytoplasmic sperm injection (ICSI), biopsy embryos and perform additional procedures in used in assisted reproductive techniques (ART), human reproduction and in vitro fertilization (IVF) techniques. The invention will allow ease in use, the reduction in the number of micro tools used in the procedure, as opposed to conventonal dishes and procedures, and will allow the user to more readily locate the oocytes and embryos to be handled and worked on. The inventon will add repetitiveness, consistency and may result in better results and outcome of the procedures. The invention will also give the user a more ergonomically correct dish for these types of procedures and related protocols.

The invention relates to a method for a silkworm diapause variety low temperature egg incubation transgene. Silkworm diapause variety next generation graine voltinism is changed in the special egg incubation condition, to ensure that present generation silkworm diapause variety moths obtained under the special egg incubation condition mate, to give birth to non-diapause next generation graine, finally referring to the microinjection transgene method of the non-diapause silkworm eggs embryo, the gene transferring is performed, therefore the obstruction of the silkworm diapause variety transgene technology is expertly broken, and a transgene silkworm is effectively manufactured. The method directly adopts the universal use utility silkworm diapause variety as original material in production, can avoid the long and troublesome breeding process, and directly utilizes the transgene technology to create the good new special silkworm variety with various characteristics, the breeding cycle can be greatly shortened, and massive manpower and physical resources are saved.

The invention discloses a method for synthetizing secretion lysozyme by middle silkgland cell of silkworm. The method comprises the following steps: building a pBSer1hLYZ (lysozyme)-A3EGFP (Enhanced Green Fluorescent Protein) plasmid for synthetizing secretion lysozyme by the silkworm; then, introducing the plasmid and an assistant plasmid capable of providing transposase into a silkworm germ cell according to the microinjection transgenosis silkworm technology; according to the transposition characteristics of a piggyBac transposon, introducing a green fluorescent protein gene and a lysozymegene into a silkworm gene group, to obtain stable heredity and expression so as to create the transgenosis silkworm capable of synthetizing secretion lysozyme by middle silkgland cell of silkworm by specificity; further, hybridizing the transgenosis silkworm with sericin silkworm; carrying out back crossing on the hybridized descendant with the sericin silkworm for 3-5 generations; and finally, carrying out selfing on the obtained product to carry out homozygosis on the lysozyme gene so as to obtain the new species of the transgenosis silkworm of the secretion lysozyme. According to the method, a basis for improving lysozyme production efficiency and lowering production cost is laid.

The invention discloses a method for improving the pig cloning efficiency on the basis of inhibition of H3K9me3 methylation. According to the method, H3K9me3 methylation is inhibited by inhibiting expression of Suv39H1 and Suv39H2 genes in nuclear donor cells during nuclear transplantation of a cloned embryo of a pig and overexpression of KDM4A mRNA during nuclear transplantation of a reconstructed embryo, and the pig cloning efficiency is further improved. Specifically, according to the method for improving the pig cloning efficiency, effective siRNA resisting Suv39H1 and Suv39H2 genes is co-transfected in the nuclear donor cells before nuclear transplantation, overexpression of the Suv39H1 and Suv39H2 genes in the nuclear donor cells is inhibited, accordingly, the high methylation level of H3K9me3 in the nuclear donor cells is inhibited, the processed nuclear donor cells are subjected to nuclear transplantation, cytoplasm microinjection of KDM4A mRNA is performed after the reconstructed embryo obtained after nuclear transplantation is activated, methylation of histone H3K9me3 in the reconstructed embryo is further inhibited, and the cloning efficiency of the pig is finally improved.