公开(公告)号：US20140295430A1

公开(公告)日：2014-10-02

申请号：US14349520

申请日：2012-10-04

Applicant: HITACHI HIGH-TECHNOLOGIES CORPORATION

Inventor： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

IPC: C12Q1/68 ,  G01N33/543

CPC classification number: C12Q1/6834 ,  C12Q1/6804 ,  C12Q1/6837 ,  G01N21/6428 ,  G01N21/6452 ,  G01N33/54326 ,  G01N33/54346 ,  G01N33/54393 ,  G01N33/587 ,  G01N2021/6421 ,  C12Q2525/161 ,  C12Q2545/114 ,  C12Q2563/107 ,  C12Q2563/143 ,  C12Q2563/149 ,  C12Q2565/514 ,  C12Q2563/119

Abstract: The method for analyzing biomolecules, includes the steps of: immobilizing biomolecules to be analyzed on surfaces of magnetic microparticles; reacting labeled probe molecules with the biomolecules to be analyzed; collecting and immobilizing the microparticles on a support substrate; and measuring a label on the support substrate. Since single-molecule immobilized magnetic microparticles are used in the present invention, the number of biomolecules can be counted, and since hybridization and an antigen-antibody reaction are performed with the microparticles having biomolecules immobilized thereon dispersed, the reaction can be rapidly performed. Further, the type and the abundance of biomolecules of interest can be determined at a single molecular level, so as to evaluate, in particular, the absolute concentration of biomolecules.

Abstract translation: 分析生物分子的方法包括以下步骤：将待分析的生物分子固定在磁性微粒的表面上; 使标记的探针分子与待分析的生物分子反应; 将微粒收集并固定在支撑基底上; 并测量支撑衬底上的标签。 由于在本发明中使用单分子固定化磁性微粒，因此可以计数生物分子的数量，由于在其上分散固定有生物分子的微粒进行杂交和抗原 - 抗体反应，所以可以快速进行反应。 此外，感兴趣的生物分子的类型和丰度可以在单个分子水平上确定，以便特别地评估生物分子的绝对浓度。

公开(公告)号：US09822399B2

公开(公告)日：2017-11-21

申请号：US14349520

申请日：2012-10-04

Applicant: HITACHI HIGH-TECHNOLOGIES CORPORATION

Inventor： Toshiro Saito ,  Koshin Hamasaki ,  Satoshi Takahashi ,  Muneo Maeshima ,  Kyoko Imai ,  Kazumichi Imai ,  Ryuji Tao

IPC: C12Q1/68 ,  G01N21/64 ,  G01N33/543 ,  G01N33/58

CPC classification number: C12Q1/6834 ,  C12Q1/6804 ,  C12Q1/6837 ,  G01N21/6428 ,  G01N21/6452 ,  G01N33/54326 ,  G01N33/54346 ,  G01N33/54393 ,  G01N33/587 ,  G01N2021/6421 ,  C12Q2525/161 ,  C12Q2545/114 ,  C12Q2563/107 ,  C12Q2563/143 ,  C12Q2563/149 ,  C12Q2565/514 ,  C12Q2563/119

Abstract: The method for analyzing biomolecules, includes the steps of: immobilizing biomolecules to be analyzed on surfaces of magnetic microparticles; reacting labeled probe molecules with the biomolecules to be analyzed; collecting and immobilizing the microparticles on a support substrate; and measuring a label on the support substrate. Since single-molecule immobilized magnetic microparticles are used in the present invention, the number of biomolecules can be counted, and since hybridization and an antigen-antibody reaction are performed with the microparticles having biomolecules immobilized thereon dispersed, the reaction can be rapidly performed. Further, the type and the abundance of biomolecules of interest can be determined at a single molecular level, so as to evaluate, in particular, the absolute concentration of biomolecules.

公开(公告)号：US09365891B2

公开(公告)日：2016-06-14

申请号：US14642461

申请日：2015-03-09

Applicant: HITACHI HIGH-TECHNOLOGIES CORPORATION

Inventor： Koshin Hamasaki ,  Toshiro Saito

IPC: C12Q1/68 ,  G01N21/64 ,  G01N33/543 ,  G01N21/552 ,  G01N21/05

CPC classification number: C12Q1/6806 ,  C12Q1/6834 ,  G01N21/05 ,  G01N21/554 ,  G01N21/648 ,  G01N33/54346 ,  C12Q2565/518 ,  C12Q2565/507 ,  C12Q2563/155

Abstract: Disclosed is a technique for binding microparticles to patterned bonding pads of a metal (e.g., gold) formed on a support. The microparticles each carry a nucleic acid synthetase or DNA probe immobilized thereon for capturing a nucleic acid sample fragment. The technique involves forming, on a support surface, a film having a thickness equivalent to that of the bonding pads; controlling the size of microparticles with respect to the size of bonding pads; and thereby immobilizing microparticles each bearing a single nucleic acid sample fragment to the bonding pads in a one-to-one manner in a grid form. This allows high-density regular alignment and immobilization of many types of nucleic acid fragment samples on a support and enables high-throughput analysis of nucleic acid samples. Typically, immobilization of microparticles at 1-micrometer intervals easily provides a high density of 106 nucleic acid fragments per square millimeter.

Abstract translation: 公开了将微粒与形成在载体上的金属（例如金）的图案化接合焊盘结合的技术。 微粒各自携带固定在其上的核酸合成酶或DNA探针，用于捕获核酸样品片段。 该技术涉及在支撑表面上形成具有与焊盘的厚度相当的厚度的膜; 控制微粒相对于焊盘尺寸的尺寸; 从而将具有单个核酸样品片段的微粒以网格形式以一对一的方式固定到接合焊盘。 这允许将许多类型的核酸片段样品高密度定期排列并固定在支持物上，并且能够进行核酸样品的高通量分析。 通常，以1微米间隔固定微粒容易提供高密度的每平方毫米106个核酸片段。

公开(公告)号：US10802017B2

公开(公告)日：2020-10-13

申请号：US14768771

申请日：2014-02-03

Applicant: HITACHI HIGH-TECHNOLOGIES CORPORATION

Inventor： Koshin Hamasaki ,  Toshiro Saito

IPC: G01N21/64 ,  G01N33/543 ,  B01L3/00 ,  G01N27/74

Abstract: Paramagnetic fine particles of 1 micron or less used under a strong magnetic field were shown to form beads-like aggregates along the magnetic flux, and become irregularly shaped as such a mass of particles combines with a flat particle layer. This phenomenon becomes a factor that degrades the quality of quantification in bioanalysis. By confining a solution of microscopic magnetic fine particles between flat substrates of high wettability as thin a vertical thickness as possible and attracting the magnetic fine particles under a magnetic field applied from the side of one of the flat substrates, the magnetic fine particles can be evenly immobilized in the form of a film on the substrate surface in a dispersion state, and the quality of the biomolecule quantification can be improved.

公开(公告)号：US10725062B2

公开(公告)日：2020-07-28

申请号：US15746434

申请日：2016-07-11

Applicant: Hitachi High-Technologies Corporation

Inventor： Koshin Hamasaki ,  Yoshihiro Yamashita

IPC: G01N1/00 ,  G01N35/10 ,  B01L3/00 ,  F16K7/14

Abstract: An amount of gas remaining within a fluid control valve is reduced according to a method for fixing the fluid control valve to achieve highly accurate trace dispensation by simply removing gas. The dispensing device has a discharge nozzle, a liquid feeding tube that is disposed so as to connect a reagent bottle in which a reagent is stored and the discharge nozzle and forms a reagent flow path, and a fluid control valve that is disposed on the liquid feeding tube route connecting the reagent bottle and the discharge nozzle. The fluid control valve is provided with a reagent flow path having a liquid inlet and a liquid outlet and a diaphragm valve provided in the middle of the flow path. The fluid control valve is disposed in an orientation such that the diaphragm valve is disposed at the bottom of the flow path of the fluid control valve.

公开(公告)号：US09964539B2

公开(公告)日：2018-05-08

申请号：US14412949

申请日：2013-06-12

Applicant: Hitachi High-Technologies Corporation

Inventor： Koshin Hamasaki ,  Toshiro Saito

IPC: G01N33/543 ,  G01N21/64 ,  C12Q1/68

CPC classification number: G01N33/54333 ,  C12Q1/6834 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/6489 ,  G01N33/54326 ,  G01N33/54346 ,  G01N2021/6441

Abstract: Biomolecules are specifically captured with magnetic particles and the biomolecules are labeled with fluorescence. A magnetic field generator, for attracting the magnetic particles to the support substrate, is provided on the reverse face of the support substrate, and an adhesion layer is provided on the surface of the support substrate to hold the magnetic particles. First, a dispersing solution for the magnetic particles is placed on the surface of the support substrate with the magnetic field in an off state. Next, the magnetic field is turned on, and the magnetic particles in solution are attracted to the support substrate surface. The magnetic particles colliding with the support substrate adhere to the adhesion layer of the support substrate surface, and then the magnetic field is turned off. Thus, aggregations can be broken up while the magnetic particles are held, and a magnetic particle layer on the support substrate can be a single layer.

公开(公告)号：US20150184227A1

公开(公告)日：2015-07-02

申请号：US14642461

申请日：2015-03-09

Applicant: HITACHI HIGH-TECHNOLOGIES CORPORATION

Inventor： Koshin Hamasaki ,  Toshiro Saito

IPC: C12Q1/68

CPC classification number: C12Q1/6806 ,  C12Q1/6834 ,  G01N21/05 ,  G01N21/554 ,  G01N21/648 ,  G01N33/54346 ,  C12Q2565/518 ,  C12Q2565/507 ,  C12Q2563/155

Abstract: Disclosed is a technique for binding microparticles to patterned bonding pads of a metal (e.g., gold) formed on a support. The microparticles each carry a nucleic acid synthetase or DNA probe immobilized thereon for capturing a nucleic acid sample fragment. The technique involves forming, on a support surface, a film having a thickness equivalent to that of the bonding pads; controlling the size of microparticles with respect to the size of bonding pads; and thereby immobilizing microparticles each bearing a single nucleic acid sample fragment to the bonding pads in a one-to-one manner in a grid form. This allows high-density regular alignment and immobilization of many types of nucleic acid fragment samples on a support and enables high-throughput analysis of nucleic acid samples. Typically, immobilization of microparticles at 1-micrometer intervals easily provides a high density of 106 nucleic acid fragments per square millimeter.

Abstract translation: 公开了将微粒与形成在载体上的金属（例如金）的图案化接合焊盘结合的技术。 微粒各自携带固定在其上的核酸合成酶或DNA探针，用于捕获核酸样品片段。 该技术涉及在支撑表面上形成具有与焊盘的厚度相当的厚度的膜; 控制微粒相对于焊盘尺寸的尺寸; 从而将具有单个核酸样品片段的微粒以网格形式以一对一的方式固定到接合焊盘。 这允许将许多类型的核酸片段样品高密度定期排列并固定在支持物上，并使核酸样品能够进行高通量分析。 通常，以1微米间隔固定微粒容易提供高密度的每平方毫米106个核酸片段。

公开(公告)号：US20150153335A1

公开(公告)日：2015-06-04

申请号：US14412949

申请日：2013-06-12

Applicant: Hitachi High-Technologies Corporation

Inventor： Koshin Hamasaki ,  Toshiro Saito

IPC: G01N33/543 ,  C12Q1/68

CPC classification number: G01N33/54333 ,  C12Q1/6834 ,  G01N21/6428 ,  G01N21/6458 ,  G01N21/6489 ,  G01N33/54326 ,  G01N33/54346 ,  G01N2021/6441

Abstract: Biomolecules are specifically captured with magnetic particles and the biomolecules are labeled with fluorescence. A magnetic field generator, for attracting the magnetic particles to the support substrate, is provided on the reverse face of the support substrate, and an adhesion layer is provided on the surface of the support substrate to hold the magnetic particles. First, a dispersing solution for the magnetic particles is placed on the surface of the support substrate with the magnetic field in an off state. Next, the magnetic field is turned on, and the magnetic particles in solution are attracted to the support substrate surface. The magnetic particles colliding with the support substrate adhere to the adhesion layer of the support substrate surface, and then the magnetic field is turned off. Thus, aggregations can be broken up while the magnetic particles are held, and a magnetic particle layer on the support substrate can be a single layer.

Abstract translation: 生物分子用磁性颗粒特异性捕获，生物分子用荧光标记。 用于将磁性颗粒吸引到支撑基板的磁场发生器设置在支撑基板的背面上，并且在支撑基板的表面上设置粘合层以保持磁性颗粒。 首先，将磁性粒子的分散液置于支撑基板的表面，使磁场成为断开状态。 接下来，磁场被打开，溶液中的磁性颗粒被吸引到支撑衬底表面。 与支撑基板碰撞的磁性粒子粘附在支撑基板表面的粘合层上，然后关闭磁场。 因此，在保持磁性颗粒的同时可以破坏聚集，并且支撑基板上的磁性颗粒层可以是单层。