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Microinjection method and deviceRelated Patent Categories: Chemistry: Molecular Biology And Microbiology, Process Of Mutation, Cell Fusion, Or Genetic Modification, Introduction Of A Polynucleotide Molecule Into Or Rearrangement Of Nucleic Acid Within An Animal CellMicroinjection method and device description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20070087436, Microinjection method and device. Brief Patent Description - Full Patent Description - Patent Application Claims
TECHNICAL FIELD [0001] The present invention relates to a method for introducing a physiologically active substance into cells and a microinjection device used for the above method. BACKGROUND ART [0002] Examples of a technique of introducing gene DNA into cultured cells or the like may include the calcium precipitation method, the lipid transfer method, the viral vector method, electroporation, the gene gun method, and the microinjection method. In the above methods other than the microinjection method, DNA is introduced in cells at a certain probability, and thus it is impossible to introduce DNA into only a specific cell. On the other hand, the microinjection method has been problematic in that since the diameter of the edge of a glass pipette is approximately 1 .mu.m, cells are easily damaged when such a glass pipette is inserted into the cell nucleus thereof. In addition, when different genes are introduced into multiple cells, the same number of pipettes as that of genes should be prepared, resulting in complicated preparation. [0003] Japanese Patent Application Laid-Open No. 2003-88383 discloses that in order to provide a means for collecting biomolecules such as RNA from living cells, a needle capable of specifically binding to biomolecules is inserted into a living cell using a device enabling fine position control, and that the needle is then removed from the cell. As a needle used herein, a ZnO whisker or a carbon nanotube is used. For example, the surface of a metal oxide whisker is modified with an amino group, so that the whisker can bind to biomolecules existing in cells and collect them. DISCLOSURE OF THE INVENTION [0004] As mentioned above, the conventional electroporation or gene gun is able to inject a substance into large quantities of cells at a time. However, it has been difficult to inject a substance into a specific cell. Moreover, the conventional microinjection is able to inject a substance into a specific cell. However, since a hollow glass capillary has been used as a needle to be injected, there has been a certain limit regarding reduction in the external diameter thereof. Thus, these conventional methods have been problematic in that a cell bursts or suffers fatal damage when a needle is injected therein, or in that operations become complicated. [0005] As described in Japanese Patent Application Laid-Open No. 2003-88383, it is possible to collect biomolecules from living cells by performing specific modification on a metal oxide whisker or a carbon nanotube. It is also possible to successively record a change in each of the cells over time. However, a method for successively recording the change in such a cell over time by actively introducing a gene therein has not yet been disclosed. In addition, the aforementioned method has been problematic in that the method comprises a complicated step of modifying the surface of a needle with a substance that is allowed to specifically bind to biomolecules. [0006] It is an object of the present invention to solve the aforementioned problems of the prior art techniques. In other words, it is an object of the present invention 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. [0007] As a result of intensive studies directed towards achieving the aforementioned object, the present inventors have found that the object can be achieved by using a needle having a diameter of 500 nm or less, provided that it is able to be inserted into a cell, and by inserting the above-described needle into the cell, thereby completing the present invention. [0008] Thus, 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. [0009] Preferably, a needle having a diameter between 50 and 100 nm, provided that it is able to be inserted into a cell, is used. [0010] Preferably, a needle having a length of 5 .mu.m or less is used. [0011] Preferably, a needle having a taper form, provided that it is able to be inserted into a cell, is used. [0012] Preferably, a needle composed of a carbon nanotube is used. [0013] Preferably, a needle composed of silicon is used. [0014] Preferably, a needle composed of a metal oxide is used. [0015] Preferably, a needle having a diameter between 50 and 500 nm, provided that it is able to be inserted into a cell, has electrical conductivity. [0016] Preferably, the physiologically active substance is DNA, RNA, or a protein. [0017] Preferably, using a needle charged with an electrical charge opposite to that of a physiologically active substance, the physiologically active substance is allowed to electrostatically attach to the above-described needle, and the above-described needle is then inserted into a cell. [0018] Preferably, using a needle to which a voltage opposite to the charge of a physiologically active substance has been applied, the physiologically active substance is allowed to electrically attach to the above-described needle, and the above-described needle is then inserted into a cell. [0019] Preferably, after a negatively charged physiologically active substance has been allowed to electrostatically attach to a needle that is positively charged, the above-described needle is inserted into a cell, and the needle is then negatively charged, so that the physiologically active substance is allowed to detach from the needle. [0020] Preferably, after a negatively charged physiologically active substance has been allowed to electrostatically attach to a needle to which a positive voltage has been applied, the above-described needle is inserted into a cell, and a negative voltage is then applied to the needle, so that the physiologically active substance is allowed to detach from the needle. [0021] Preferably, negative voltages that change over time are applied to the needle, so that the physiologically active substance is allowed to detach from the needle. Continue reading about Microinjection method and device... Full patent description for Microinjection method and device Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Microinjection method and device patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. Start now! - Receive info on patent apps like Microinjection method and device or other areas of interest. ### Previous Patent Application: Multicellular compositions of pluripotent human embryonic stem cells and cancer cells Next Patent Application: Methods for rejuvenating cells in vitro and in vivo Industry Class: Chemistry: molecular biology and microbiology ### FreshPatents.com Support Thank you for viewing the Microinjection method and device patent info. IP-related news and info Results in 0.11088 seconds Other interesting Feshpatents.com categories: Computers: Graphics , I/O , Processors , Dyn. Storage , Static Storage , Printers 174 |
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