430 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.

This invention provides methods for the stable introduction of heterologous coding sequences into the genome of a bird and expressing the coding sequences to produce desired proteins or to alter the phenotype of the bird. The present invention provides preferred methods for introducing a transgene into the cytoplasm of avian embryonic cells by cytoplasmic microinjection. The embryo then develops into a transgenic adult capable of expressing a heterologous protein and / or capable of generating a line of transgenic birds through breeding. Synthetic vectors and gene promoters useful in the methods are also provided by the present invention, as are transgenic birds that express heterologous protein and avian eggs containing heterologous protein.

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 provides a breeding method for knocking out fish MC4R gene by using a CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 system. The method comprises the following steps: determination of MC4R gene targeted site, preparation of gRNA, in-vitro microinjection and knock-out verification, screening of MC4R-gene-knock-out fish, culture of hybrid fish, and the like. The breeding method is suitable for all commercial fishes, and can implement quick growth and breeding of fishes by knocking out the fish MC4R gene. Compared with the traditional breeding process, the method provided by the invention has the characteristics of high accuracy, low cost and short pure line acquisition time. Compared with the transgenic breeding, the gene knock-out can cause gene malfunction of the fish, and can not bring in any external gene, thereby avoiding the problem of transgenic safety.

A method has been developed to produce stable cell-matrix microspheres with up to 100% encapsulation efficiency and high cell viability, using matrix or biomaterial systems with poor shape and mechanical stability for applications including cell therapeutics via microinjection or surgical implantation, 3D culture for in vitro expansion without repeated cell splitting using enzymatic digestion or mechanical dissociation and for enhanced production of therapeutic biomolecules, and in vitro modeling for morphogenesis studies. The modified droplet generation method is simple and scalable and enables the production of cell-matrix microspheres when the matrix or biomaterial system used has low concentration, with slow phase transition, with poor shape and mechanical stability.

A microinjection device is provided that includes an injection element defining a longitudinal axis, and that further includes a motor. The injection element is rotatable about the longitudinal axis by the rotational motor. The injection element is for penetrating a target, such as a cell. A microinjection system is provided that includes the microinjection device and a control unit. The control unit is for controlling a rotational amplitude and a frequency of oscillation of the injection element. A method for penetrating a target to facilitate injecting material therein is provided that includes providing the material to an injection element, contacting the target with a distal end of the injection element, rotating the injection element about a longitudinal axis to form a hole in the target, and penetrating the target with the injection element via the hole formed in the target. A method for performing intra-cytoplasmic sperm injection is provided that includes providing a solution comprising sperm to an injection element, contacting an oocyte with a distal end of the injection element, rotating the injection element alternately clockwise and counterclockwise about a longitudinal axis to form a hole in the oocyte, penetrating the oocyte with the distal end of the injection element via the hole formed in the oocyte, and expelling the solution comprising sperm into the penetrated oocyte.

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 belongs to the field of fish molecular breeding, and particularly relates to a fish breeding method for improving CRISPR-Cas9 gene editing and passage efficiency through fish roe preserving fluid. According to the method, the fish roe preserving fluid technology, the microinjection technology an the CRISPR-Cas9 gene editing technology are ingeniously combined, the targeting efficiency of the CRISPR-Cas9 gene editing technology and gene editing passage efficiency are greatly improved, the screening time needed by a gene editing method for fish breeding is obviously saved, and the method is of great significance in promoting rapid development of gene editing fish breeding.

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 double-circular-ring-shaped slit type electrostatic spinning device and method. The electrostatic spinning device comprises a liquid supply system, a double-circular-ring-shaped slit type nozzle system and a nano fiber collection system. The electrostatic spinning method comprises the following steps of: starting a microinjection pump to enable a solution to flow into a slit between an inner ring and an outer ring through a micro-channel in an inner core; adjusting the flow of the solution to enable a liquid surface of the solution to fully overflow in the slit, but not flow out; starting a speed regulating motor to enable a metal roller to rotate normally; starting a high-voltage electrostatic generator and slowly increasing voltage; when the voltage exceeds a certain value, forming a plurality of jet flows in the slit; in a process that the jet flows fly to the metal roller, volatilizing a solvent and curing a high polymer to form nano fibers which are deposited on the metal roller; and rotating the metal roller to collect the nano fibers. By adopting the microinjection pump and the liquid supply system, a high polymer solution can be automatically and accurately supplied, and the solution is sufficiently and efficiently utilized, so that the yield of the nano fibers is improved and continuous preparation of the nano fibers is realized.

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 microscopic cell injecting instrument includes cell conveyer, cell injector, cell holder and digital control and drive system. In cell injection, cell is conveyed automatically with the cell conveyer and in microfluid conveying mode to the cell injector, and the outlet of the cell holder. During cell injection, all the cell conveyance, holding, cell membrane puncture with the injection needle, intracellular medicine injection, the withdrawn of injection needle, cell release and other microscopic operation steps are computer digital controlled in the same computer operating interface. The present invention has simple operation, high successful injection rate, and automatic separation and counting of injected and un-injected cells.

The present invention relates generally to microelectromechanical systems (MEMS) devices for the manipulation of cells or groups of cells, such as oocytes, embryos, and sperm. In particular, the present invention relates to Cell Labeling MEMS devices (2F), Microinjection MEMS devices, IntraCytoplasmic Sperm Injection (“ICSI”) MEMS devices, Zona Coring MEMS devices, Enucleation MEMS devices, Enucleation / Nuclear Transfer MEMS devices, and Cytoplasmic Transfer MEMS devices. The present invention also relates to kits containing the MEMS devices of the present invention.

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.

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 includes reagents and methods for the isolation of nucleic acids. The reagents described herein contain a nucleic acid precipitating agent and a solid phase carrier. The reagents can optionally be formulated to cause the lysis of a cell. These reagents can be used to isolate a target nucleic acid molecule from a cell or a solution containing a mixture of different size nucleic acid molecules. The disclosed reagents and methods provides a simple, robust and readily automatable means of nucleic acid isolation and purification which produces high quality nucleic acid molecules suitable for: capillary electrophoresis, nucleotide sequencing, reverse transcription cloning the transfection, transduction or microinjection of mammalian cells, gene therapy protocols, the in vitro synthesis of RNA probes, cDNA library construction and PCR amplification.

The invention discloses a screw-free ultrasonic microinjection molding device for a molten and plasticized polymer and a molding method. An injection part comprises an injection motor, a lead screw, an injection push plate and a connecting rod; an ultrasonic vibration system comprises an ultrasonic generator, an energy converter, an amplitude-change pole and a tool head; an injection module is provided with a plasticizing hole, a sub-runner and a mould cavity; a feeding system comprises a hopper and a support; and a mould closing and ejecting system comprises a mould closing hydraulic cylinder, an ejecting hydraulic cylinder, an upper mould fixing plate, a lower mould fixing plate and an ejector rod. A solid polymer is molten into a liquid under the action of frication thermogenesis and viscoelasticity thermogenesis applied into the polymer particle process through ultrasonic vibration, and meanwhile, ultrasonic waves play a cavitation role in the polymer liquid, so that the apparent viscosity is reduced, the phenomenon of short shot or insufficient filling in the machining process is overcome, the injection molding of a small plastic device can be well realized, the energy consumption is reduced, the difficulty in controlling the process is lowered, the economic efficiency of the device is increased, and the production cost is reduced.

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 present invention relates to a method using a cell penetrating peptide (Pep-1) for labeling and delivering mitochondria separated from healthy cells to replace damaged mitochondria. At present, microinjection of mitochondria into cells can only process one cell at a time, and therefore, this technique is limited to embryo related research and relevant applications. The advantages of the said peptide-mediated mitochondrial delivery system (PMD) include less steps with more efficiency, where a number of cells can be treated following one labeling process; the delivery process can be easily controlled, there is no cell toxicity after delivery under appropriate conditions, and delivery efficiency is over 80% depending on different cell types. Mitochondria delivered by the PMD system will move to the original mitochondrial location in the cells and will not be catalyzed in lysosomes; thus, the therapeutic effects can last at least one week.

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.