公开(公告)号：US11053536B2

公开(公告)日：2021-07-06

申请号：US14418135

申请日：2012-07-30

申请人： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi ,  Maiko Tanabe

发明人： Masataka Shirai ,  Hideki Kambara ,  Kiyomi Taniguchi ,  Maiko Tanabe

IPC分类号： C12Q1/6837 ,  C12N15/10

摘要： The present invention relates to a method, a device, and an apparatus for analyzing the expression of a gene in single cells. Specifically, the present invention relates to: a device for gene expression analysis, characterized by including a support, in which a nucleic acid probe having a test nucleic acid capture sequence and a known sequence, and further containing a cell recognition tag sequence which differs depending on the difference in position on the surface of the support or in the vicinity of the surface thereof, and a common primer sequence having a known sequence is two-dimensionally distributed and immobilized on the surface of the support or in the vicinity of the surface thereof; and a method and an apparatus using the device for gene expression analysis.

公开(公告)号：US09278355B2

公开(公告)日：2016-03-08

申请号：US12292167

申请日：2008-11-13

申请人： Tomoharu Kajiyama ,  Masataka Shirai ,  Hideki Kambara

发明人： Tomoharu Kajiyama ,  Masataka Shirai ,  Hideki Kambara

IPC分类号： C12N15/10 ,  C12Q1/68 ,  B01L3/00

CPC分类号： B01L3/50851 ,  B01L3/5025 ,  B01L3/50857 ,  B01L2200/0668 ,  B01L2300/0819

摘要： This invention provides a nucleic acid amplification device whereby the abundance of a target molecule can be maintained before and after a step of separately amplifying a sample such that highly accurate analysis results that can be applied to gene expression analysis can be obtained. Also, a nucleic acid amplification device having a structure in which a plurality of minute reaction cells each comprising a set of a bead-retaining space capable of retaining a single analysis bead and a reagent reaction space retaining no bead but having a volume that is large enough to cause a reagent reaction therein are positioned so as to form a planar face is provided.

公开(公告)号：US20120270210A1

公开(公告)日：2012-10-25

申请号：US13389842

申请日：2010-08-10

申请人： Masayasu Kuwahara ,  Tomoharu Kajiyama ,  Hideki Kambara

发明人： Masayasu Kuwahara ,  Tomoharu Kajiyama ,  Hideki Kambara

IPC分类号： C12Q1/68 ,  C07H19/14

CPC分类号： C07H19/20 ,  C12Q1/6869 ,  C12Q2525/117 ,  C12Q2565/301 ,  C12Q2525/113

摘要： Provided is a nucleic acid substrate which has nucleic acid substrate characteristics equivalent to those of dATP, has a low substrate specificity for luciferase, exerts no negative effect on enzymatic reactions such as a complementary-strand synthesis, and therefore is particularly suitable for the pyrosequencing method. As a nucleic acid substrate complementary to nucleotide T, a 7-substituted deoxyribonucleotide triphosphate whose 7-position of a purine group is modified by a substituent is used as a substitute for a nucleotide α-thiotriphosphate analog.

摘要翻译： 提供具有与dATP相同的核酸底物特征的核酸底物，对荧光素酶具有低底物特异性，对互补链合成等酶反应没有不利影响，因此特别适用于焦磷酸测序法 。 作为与核苷酸T互补的核酸底物，使用嘌呤基的7-位通过取代基修饰的7-取代的脱氧核糖核苷酸三磷酸作为核苷酸α-硫代三磷酸类似物的替代物。

公开(公告)号：US08231828B2

公开(公告)日：2012-07-31

申请号：US11601725

申请日：2006-11-20

申请人： Tomoharu Kajiyama ,  Hideki Kambara ,  Kunio Harada

发明人： Tomoharu Kajiyama ,  Hideki Kambara ,  Kunio Harada

IPC分类号： G01N21/00

CPC分类号： B01L3/0241 ,  B01J2219/00376 ,  B01J2219/00378 ,  B01J2219/00416 ,  B01J2219/00418 ,  B01J2219/00484 ,  B01L2200/0621 ,  B01L2200/16 ,  B01L2300/0838 ,  B01L2400/0487 ,  B01L2400/049 ,  G01N35/1072 ,  Y10T436/2575

摘要： By the conventional technique for dispensing more than one reagents accurately, the system is complicated and thus a compact and inexpensive system is difficult to realize. In the present invention, the pressurized dispensing system utilizing a capillary is realized, and in addition, in order to reduce the leakage of reagents different from the reagent dispensed, by forming air layers at the tips of the capillaries after dispensing, a compact, simple, inexpensive analysis apparatus is realized.

公开(公告)号：US20080260577A1

公开(公告)日：2008-10-23

申请号：US12032091

申请日：2008-02-15

申请人： Masataka SHIRAI ,  Tomoharu Kajiyama ,  Hideki Kambara

发明人： Masataka SHIRAI ,  Tomoharu Kajiyama ,  Hideki Kambara

IPC分类号： G01N21/76

CPC分类号： G01N21/76 ,  B01L3/502761 ,  B01L2200/0668 ,  B01L2300/0816 ,  B01L2300/0877 ,  G01N21/01 ,  G01N21/251 ,  G01N21/763

摘要： Throughput is improved by increasing the number of micro-reaction chambers. There is provided a chemiluminescent detection system that has a so-called plate on which many reaction chambers are one-dimensionally or two-dimensionally arranged, characterized in that optical detection is performed using a line or area sensor having many detection pixels, the spacing of the optical detection pixels substantially matches the spacing of the reaction chambers on the plate, and the micro-reaction chambers and the pixels are made to correspond one-to-one with each other so that light from the reaction chambers on the plate enters the detection pixels most efficiently and does not scatter to other pixels. To make the micro-reaction chambers arrayed on the plate and the pixels of the image pickup element plate correspond one-to-one with each other, light-emitting substances or reflectors or photoabsorption substances are placed so as to serve as alignment marks.

摘要翻译： 通过增加微反应室的数量来改善生产能力。 提供一种化学发光检测系统，其具有所谓的板，其中许多反应室被一维或二维排列，其特征在于，使用具有许多检测像素的线或区域传感器进行光学检测， 光学检测像素基本上与板上的反应室的间隔相匹配，并且使微反应室和像素彼此一一对应，使得来自板上的反应室的光进入检测 像素最有效，不会散射到其他像素。 为了使微反应室排列在板上并且图像拾取元件板的像素彼此一一对应，放置发光物质或反射体或光吸收物质以作为对准标记。

公开(公告)号：US07241597B2

公开(公告)日：2007-07-10

申请号：US10634795

申请日：2003-08-06

申请人： Chihiro Uematsu ,  Hideki Kambara ,  Kazunon Okano

发明人： Chihiro Uematsu ,  Hideki Kambara ,  Kazunon Okano

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/6809 ,  C12Q1/6855 ,  C12Q2525/173

摘要： The inventive method for assaying DNA fragments in mixture comprises step 1 of ligating different oligomers hybridizable to primers of the same melting temperature and the same length to individual groups of DNA fragments in a set of DNA fragments; step 2 of mixing together the groups of DNA fragments ligated with the oligomers; step 3 of simultaneous PCR of the groups of DNA fragments ligated with the oligomers in one receptacle by using the primers being complementary to the oligomers and corresponding to the individual groups; and step 4 of detecting PCR amplified DNA fragments; characterized in that the method enables the comparison of plural samples under no influence of PCR reproducibility.

摘要翻译： 用于测定混合物中DNA片段的本发明方法包括步骤1，将不同的低聚物连接到一组DNA片段中与相同融合温度和相同长度的引物相同的DNA片段的单个组; 步骤2，将与低聚物连接的DNA片段组合在一起; 通过使用与寡聚体互补并对应于各个基团的引物，在一个容器中与低聚物连接的DNA片段组同时PCR的步骤3; 和检测PCR扩增的DNA片段的步骤4; 其特征在于，该方法能够在不影响PCR再现性的情况下比较多个样品。

公开(公告)号：US06821446B2

公开(公告)日：2004-11-23

申请号：US09944410

申请日：2001-09-04

申请人： Kunio Harada ,  Masao Kamahori ,  Hideki Kambara ,  Sumio Yamaguchi ,  Sukeyoshi Tsunekawa

发明人： Kunio Harada ,  Masao Kamahori ,  Hideki Kambara ,  Sumio Yamaguchi ,  Sukeyoshi Tsunekawa

IPC分类号： C03C1500

CPC分类号： C03C17/005 ,  C03C17/32 ,  C03C2218/328 ,  G01N27/44704 ,  Y10T428/131 ,  Y10T428/1321 ,  Y10T428/26 ,  Y10T428/2933 ,  Y10T428/2964 ,  Y10T428/2973 ,  Y10T428/2975 ,  Y10T428/2976 ,  Y10T428/31623

摘要： The present invention provides a method for producing a high-quality capillary tube used in an electrophoresis apparatus in a safe and inexpensive manner. A polymer coating on a capillary tube is converted into gas and removed through an oxidative reaction with oxygen radicals resulting from ozone decomposition.

摘要翻译： 本发明提供一种以安全且廉价的方式生产用于电泳装置的高品质毛细管的方法。 将毛细管上的聚合物涂层转化为气体，并通过氧化反应除去臭氧分解产生的氧自由基。

公开(公告)号：US06750018B2

公开(公告)日：2004-06-15

申请号：US09945703

申请日：2001-09-05

申请人： Hideki Kambara ,  Guohua Zhou ,  Kazunori Okano ,  Masao Kamahori

发明人： Hideki Kambara ,  Guohua Zhou ,  Kazunori Okano ,  Masao Kamahori

IPC分类号： C12Q168

CPC分类号： C12Q1/6858 ,  C12Q2565/537 ,  C12Q2565/301 ,  C12Q2535/125

摘要： To provide a method and an apparatus for detecting DNA mutation without using electrophoresis, the presence or absence of specific sequence is detected by a method comprising a step of serially hybridizing a 5-base-long to 8-base-long short oligomer 6, which is capable of being engaged in the complementary strand extension reaction, and a long oligomer 61, which is capable of hybridizing with a target DNA but incapable of being engaged in the complementary strand extension reaction, with the target DNA 63; a step of carrying out the complementary strand extension reaction 66 starting from the short primer using four kinds of nucleic acid substrates and polymerase; and a step of detecting photo-emission, in which pyrophosphate 68 produced as a reaction by-product of the complementary strand extension reaction is converted into ATP and the photo-emission reaction is carried out using an enzyme.

摘要翻译： 为了提供用于在不使用电泳的情况下检测DNA突变的方法和装置，通过包括与5-碱基长至8-碱基长的短寡聚物6串联杂交的步骤的方法来检测特异性序列的存在或不存在，其中 能够进行互补链延伸反应，以及能够与靶DNA杂交但不能与互补链延伸反应进行杂交的长寡聚体61与目标DNA 63反应; 使用四种核酸底物和聚合酶从短引物开始进行互补链延伸反应66的步骤; 检测发光的步骤，其中作为互补链延伸反应的反应副产物产生的焦磷酸盐68被转化为ATP，并且使用酶进行光发射反应。

公开(公告)号：US06225064B1

公开(公告)日：2001-05-01

申请号：US09413814

申请日：1999-10-07

申请人： Chihiro Uematsu ,  Hideki Kambara ,  Kazunori Okano

发明人： Chihiro Uematsu ,  Hideki Kambara ,  Kazunori Okano

IPC分类号： C12Q168

CPC分类号： C12Q1/6809 ,  C12Q1/6855 ,  C12Q2525/173

摘要： The inventive method for assaying DNA fragments in mixture comprises step 1 of ligating different oligomers hybridizable to primers of the same melting temperature and the same length to individual groups of DNA fragments in a set of DNA fragments; step 2 of mixing together the groups of DNA fragments ligated with the oligomers; step 3 of simultaneous PCR of the groups of DNA fragments ligated with the oligomers in one receptacle by using the primers being complementary to the oligomers and corresponding to the individual groups; and step 4 of detecting PCR amplified DNA fragments; characterized in that the method enables the comparison of plural samples under no influence of PCR reproducibility.

摘要翻译： 用于测定混合物中DNA片段的本发明方法包括连接不同寡聚物的步骤，所述寡聚物与一组DNA片段中的DNA片段的相同融合温度和相同长度的引物可混合;步骤2将DNA片段组合在一起 与寡聚体连接;步骤3，通过使用与低聚物互补并对应于各个基团的引物，在一个容器中与寡聚物连接的DNA片段组同时PCR; 检测PCR扩增的DNA片段; 其特征在于，该方法能够在不影响PCR再现性的情况下比较多个样品。

公开(公告)号：US6132578A

公开(公告)日：2000-10-17

申请号：US880544

申请日：1997-06-23

申请人： Hideki Kambara ,  Keiichi Nagai ,  Hiroaki Machida

发明人： Hideki Kambara ,  Keiichi Nagai ,  Hiroaki Machida

IPC分类号： G01N21/64 ,  G01N27/447 ,  G01N27/26

CPC分类号： G01N27/44721 ,  G01N27/44782

摘要： An electrophoresis separation and detection apparatus for separating and detecting fluorophore-labeled samples by electrophoresis, which comprises a migrating and separating means for electrophoresis separation of the samples labeled with two or more kinds of fluorophore labels, respectively; an irradiating means which casts two or more kinds of exciting lights having different wavelengths on a plurality of positions in all the electrophoresis lanes or on the extensions of all the electrophoresis lanes substantially at the same time; and a detecting means which detects the samples separated by the electrophoresis, at the positions on which the exciting lights are casted, wherein each fluorophore label is composed of a substance for excitation which is excited by the exciting light and a substance for emission which emits light owing to energy transfer from the substance for excitation; two or more kinds of substances are used as each of the substance for excitation and the substance for emission; and the detecting means detects the samples by distinguishing the two or more kinds of the fluorophore labels from one another on the basis of the difference of combination of the two or more kinds of the substances used as the substance for excitation and those used as the substance for emission.

摘要翻译： 一种用于通过电泳分离和检测荧光团标记的样品的电泳分离和检测装置，其包括用于分别用两种或多种荧光标记物标记的样品进行电泳分离的迁移和分离装置; 基本上同时在所有电泳泳道或所有电泳车道的延伸部分上投射具有不同波长的两种或更多种激发光的照射装置; 以及检测装置，其检测通过电泳分离的样品，在其上投射激发光的位置，其中每个荧光标记由被激发光激发的激发物质和发出光的物质组成 由于激发物质的能量转移; 使用两种或更多种物质作为激发物质和排放物质。 检测装置根据用作激发物质的物质和用作物质的两种或更多种物质的组合的差异，通过区分两种或更多种荧光团标记物来检测样品。 用于排放。