公开(公告)号：US20140356969A1

公开(公告)日：2014-12-04

申请号：US14465208

申请日：2014-08-21

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa ,  Takuya Hanashi

IPC: G01N33/53 ,  G01N21/64 ,  G01N33/52 ,  G01N21/76

CPC classification number: G01N33/5308 ,  G01N15/1434 ,  G01N21/6486 ,  G01N21/76 ,  G01N33/52 ,  G01N33/54313 ,  G01N2201/12 ,  G02B21/00 ,  G02B21/0024 ,  G02B21/0076 ,  Y10T436/143333

Abstract: Provided is a method for detecting a target particle that is a method for detecting a non-luminescent target particle dispersed and randomly moving in a sample solution using an optical system of a confocal microscope or multi-photon microscope, having: (a) preparing a sample solution containing target particles, and labeling particles of which the average outer diameter is less than 15% of the diameter of a photodetection region of the optical system, binding two or more molecules of the labeling particles per molecule of the target particles in the sample solution, and forming a non-luminescent complex of which the outer diameter is 15% or more of the diameter of the photodetection region; and, (b) calculating the number of molecules of the complex in the sample solution prepared in the (a) using an inverse scanning molecule counting method.

Abstract translation: 本发明提供一种检测作为使用共聚焦显微镜或多光子显微镜的光学系统在样品溶液中分散并随机移动的非发光靶颗粒的方法的目标颗粒的检测方法，其特征在于：（a） 含有目标颗粒的样品溶液，并且标记其平均外径小于光学系统的光电探测区域的直径的15％的颗粒，每个分子中的每个分子的靶标颗粒结合两个或更多个分子的样品 并且形成外径为光电检测区域的直径的15％以上的非发光性络合物; 和（b）使用逆扫描分子计数法计算在（a）中制备的样品溶液中的络合物的分子数。

公开(公告)号：US20140004519A1

公开(公告)日：2014-01-02

申请号：US13946059

申请日：2013-07-19

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa ,  Hidetaka Nakata

IPC: C12Q1/68

CPC classification number: C12Q1/6818 ,  C12Q1/6827 ,  G01N21/6428 ,  G01N21/6452 ,  C12Q2535/131 ,  C12Q2543/10 ,  C12Q2545/114 ,  C12Q2563/107 ,  C12Q2565/107 ,  C12Q2565/601

Abstract: The present invention provides a method for identifying polymorphism of nucleic acids in a sample solution in which the concentration or number density of the observed nucleic acids is lower than that of conventional photometric analysis technologies. It includes: preparing a sample solution comprising a first nucleic acid probe, which specifically hybridizes with a single-stranded nucleic acid molecule including a first type of base sequence, and a target nucleic acid molecule; forming a hybrid of the nucleic acid molecules in the sample solution; calculating a number of molecules of the hybrid including the first nucleic acid probe in the sample solution by the scanning molecule counting method; and identifying polymorphism of the target nucleic acid molecule based on the calculating result. The sample solution includes an oligonucleotide having a base sequence complementary to a base sequence different from the first type of base sequence in the polymorphic sequence.

Abstract translation: 本发明提供了一种用于鉴定样品溶液中核酸多态性的方法，其中观察到的核酸的浓度或数量密度低于常规光度分析技术的浓度或数量密度。 其包括：制备包含与包含第一类碱基序列的单链核酸分子和靶核酸分子特异性杂交的第一核酸探针的样品溶液; 形成样品溶液中核酸分子的杂交体; 通过扫描分子计数法计算样品溶液中包含第一核酸探针的杂合物数目; 以及基于所述计算结果鉴定所述靶核酸分子的多态性。 样品溶液包括具有与多态性序列中与第一类型碱基序列不同的碱基序列互补的碱基序列的寡核苷酸。

公开(公告)号：US20140004518A1

公开(公告)日：2014-01-02

申请号：US13944550

申请日：2013-07-17

Applicant: Olympus Corporation

Inventor： Hidetaka Nakata ,  Kazutaka Nishikawa

IPC: C12Q1/68

CPC classification number: C12Q1/6818 ,  C12Q1/6827 ,  C12Q1/6886 ,  G01N21/6428 ,  G01N21/6452 ,  C12Q2525/301 ,  C12Q2543/10 ,  C12Q2563/107 ,  C12Q2565/1015 ,  C12Q2565/107 ,  C12Q2565/601

Abstract: The present invention provides a method for identifying polymorphism of nucleic acids in a sample solution in which the concentration or number density of the observed nucleic acids is lower than that of conventional photometric analysis technologies. Namely, the present invention relates to a method for identifying polymorphism of nucleic acid molecules, in which is compared the result of hybridization between a target nucleic acid molecule and a first nucleic acid probe labeled with a fluorescent substance; and the result of hybridization between the target nucleic acid molecule and a second nucleic acid probe having a sequence different from the sequence of the first nucleic acid probe, and being labeled with a fluorescent substance. Each of the hybridization is conducted in a separate sample solution and the detection of the conjugate is done by counting the molecule of the conjugate using a scanning molecule counting method.

Abstract translation: 本发明提供了一种用于鉴定样品溶液中核酸多态性的方法，其中观察到的核酸的浓度或数量密度低于常规光度分析技术的浓度或数量密度。 即，本发明涉及一种鉴定核酸分子多态性的方法，其中比较了靶核酸分子与用荧光物质标记的第一核酸探针之间的杂交结果; 以及靶核酸分子与具有与第一核酸探针的序列不同的序列的第二核酸探针之间的杂交结果，并用荧光物质标记。 每个杂交在单独的样品溶液中进行，并且通过使用扫描分子计数法计数缀合物的分子来进行缀合物的检测。

公开(公告)号：US20130228705A1

公开(公告)日：2013-09-05

申请号：US13788972

申请日：2013-03-07

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa ,  Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/64

CPC classification number: G01N21/64 ,  G01N15/1463 ,  G01N21/6428 ,  G01N21/6452 ,  G01N2015/1493

Abstract: There is provided a scanning molecule counting method using an optical measurement with a confocal microscope or a multiphoton microscope, enabling characterization of a light-emitting particle or identification of a light-emitting particle with emitted light intensity of a single light-emitting particle measured individually. In the inventive optical analysis technique, with reference to the ratio of the intensities of simultaneously generated signals of the lights of at least two light-emitting sites having mutually different emission wavelengths, possessed by a light-emitting particle contained in a sample solution, the intensities being measured with moving the position of the light detection region of an optical system by changing the optical path of the optical system, a single light-emitting particle corresponding to the signals is identified, and the kind, the size, etc. of the light-emitting particle is identified.

Abstract translation: 提供了一种使用共焦显微镜或多光子显微镜的光学测量的扫描分子计数方法，能够单独测量发光粒子的特征或发光粒子的发射光强度的单个发光粒子的发光强度。 。 在本发明的光学分析技术中，参照包含在样品溶液中的发光粒子具有的具有相互不同的发射波长的至少两个发光部位的至少两个发光部位的同时产生的信号的强度的比例， 通过改变光学系​​统的光路来移动光学系统的光检测区域的位置来测量强度，识别与该信号相对应的单个发光粒子，并且确定该光学系统的种类，尺寸等 发现发光粒子。

公开(公告)号：US20140147854A1

公开(公告)日：2014-05-29

申请号：US14172295

申请日：2014-02-04

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa

IPC: G01N21/64

CPC classification number: G01N21/6486 ,  G01N21/278 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/76 ,  G01N2021/6417 ,  G01N2021/6419 ,  G01N2021/6421 ,  G01N2021/6493

Abstract: This method for detecting a target particle has: (a) a step for preparing a sample solution containing target particles and one type or two or more types of a luminescent probe that binds to the target particles, and allowing two or more molecules of the luminescent probe to bind per one molecule of the target particles in the sample solution, and (b) a step for calculating the number of molecules of target particles bound to the luminescent probe present in the sample solution prepared in step (a) by a scanning molecule counting method by using as an indicator thereof the strength of light signals of the individually detected particles, and the luminescent probe is one type or two or more types of a luminescent probe to which the same type of luminescent substance is bound.

Abstract translation: 用于检测目标颗粒的方法具有：（a）制备含有目标颗粒的样品溶液和结合目标颗粒的一种或两种或更多种类型的发光探针的步骤，并且允许两个或更多个分子的发光 探针在样品溶液中每一分子的目标颗粒结合，和（b）计算通过扫描分子计算与步骤（a）中制备的样品溶液中存在的发光探针结合的目标颗粒的分子数目的步骤 通过使用单独检测的颗粒的光信号的强度作为指示器的计数方法，并且发光探针是与相同类型的发光物质结合的发光探针的一种或两种以上。

公开(公告)号：US11906410B2

公开(公告)日：2024-02-20

申请号：US17135369

申请日：2020-12-28

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa ,  Jun Funazaki

IPC: G01N1/40 ,  C12Q1/04 ,  C12Q1/24 ,  G01N1/30

CPC classification number: G01N1/4077 ,  C12Q1/04 ,  C12Q1/24 ,  G01N1/30 ,  G01N2001/4088

Abstract: A cell inspection method includes a concentrate production step, a staining step, a cell precipitation step, and an observation step. In the concentrate production step, the cell concentrate is produced by causing an inner cylinder which has a filter provided on a bottom surface to enter from the bottom surface side into the through hole of the outer cylinder and bringing the inner cylinder closer to the slide, the inner cylinder having an internal space, and in the observation step, observation is performed in a state where the inner cylinder is entered into the outer cylinder.

公开(公告)号：US09354176B2

公开(公告)日：2016-05-31

申请号：US14172295

申请日：2014-02-04

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa

IPC: C12Q1/68 ,  G01N21/64 ,  G01N21/27 ,  G01N21/76

CPC classification number: G01N21/6486 ,  G01N21/278 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/76 ,  G01N2021/6417 ,  G01N2021/6419 ,  G01N2021/6421 ,  G01N2021/6493

Abstract: This method for detecting a target particle has: (a) a step for preparing a sample solution containing target particles and one type or two or more types of a luminescent probe that binds to the target particles, and allowing two or more molecules of the luminescent probe to bind per one molecule of the target particles in the sample solution, and (b) a step for calculating the number of molecules of target particles bound to the luminescent probe present in the sample solution prepared in step (a) by a scanning molecule counting method by using as an indicator thereof the strength of light signals of the individually detected particles, and the luminescent probe is one type or two or more types of a luminescent probe to which the same type of luminescent substance is bound.

Abstract translation: 用于检测目标颗粒的方法具有：（a）制备含有目标颗粒的样品溶液和结合目标颗粒的一种或两种或更多种类型的发光探针的步骤，并且允许两个或更多个分子的发光 探针在样品溶液中每一分子的目标颗粒结合，和（b）计算通过扫描分子计算与步骤（a）中制备的样品溶液中存在的发光探针结合的目标颗粒的分子数目的步骤 通过使用单独检测的颗粒的光信号的强度作为指示器的计数方法，并且发光探针是与相同类型的发光物质结合的发光探针的一种或两种以上。

公开(公告)号：US20140179023A1

公开(公告)日：2014-06-26

申请号：US14189197

申请日：2014-02-25

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa

IPC: C12Q1/68

CPC classification number: C12Q1/6825 ,  G01N15/1456 ,  G01N21/6408 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N2015/1006 ,  G01N2021/6419 ,  G01N2021/6421 ,  G01N2021/6432 ,  G02B21/0076

Abstract: Provided is a method for detecting a target particle in a biosample containing pancreatic juice, the method enabling the detection in a solution that has a lower concentration or number density of the target particles than the level possible for conventional photoanalysis techniques. This method comprises: a probe-binding step for preparing a sample solution, which contains a biosample containing pancreatic juice and a fluorescent probe capable of binding to a target particle, and binding the fluorescent probe to the target particle in the biosample; and a calculation step for calculating the number of molecules of the target particles bound to the fluorescent probes by the scanning molecule counting method. A light emission property of emitted light is different between a state where the fluorescent probe is bound to the target particle and a state where the fluorescent probe is present alone. In a state where the fluorescent probe is bound to the target particle, the fluorescent probe emits fluorescence having a wavelength of 600 nm or longer.

Abstract translation: 提供了一种用于检测含有胰液的生物样品中的靶颗粒的方法，该方法能够在具有比目标颗粒更低的浓度或数量密度的溶液中进行检测，而不是常规光分析技术可能的水平。 该方法包括：用于制备样品溶液的探针结合步骤，其包含含有胰液的生物样品和能够结合靶颗粒的荧光探针，并将荧光探针与生物样品中的靶颗粒结合; 以及通过扫描分子计数法计算与荧光探针结合的靶粒子的分子数的计算步骤。 发光的发光特性在荧光探针与目标粒子结合的状态和荧光探针单独存在的状态之间是不同的。 在荧光探针与目标粒子结合的状态下，荧光探针发出波长为600nm以上的荧光。

公开(公告)号：US20140093874A1

公开(公告)日：2014-04-03

申请号：US14056123

申请日：2013-10-17

Applicant: OLYMPUS CORPORATION

Inventor： Kazutaka Nishikawa

IPC: C12Q1/68

CPC classification number: C12Q1/6816 ,  C12Q1/6818 ,  G01J3/4406 ,  G01N15/1456 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6458 ,  G01N2015/0038 ,  G01N2015/1486 ,  G01N2021/6419 ,  G01N2021/6421 ,  G02B21/0076 ,  C12Q2543/10 ,  C12Q2545/114 ,  C12Q2565/601

Abstract: A method for detecting a nucleic acid molecule comprises: preparing a sample solution which contains a nucleic acid probe which is specifically hybridizable with the nucleic acid molecule to be analyzed, and a biosample; associating the nucleic acid molecule with the nucleic acid probe in the sample solution that has been prepared in the preparing; and after the associating, calculating, by the scanning molecule counting method, the number of molecules of the associated bodies including the nucleic acid probe in the sample solution that has been prepared in the preparing. This method for detecting a nucleic acid molecule further comprises: removing proteins from the sample solution before the calculating, or removing proteins from the biosample before the preparing.

Abstract translation: 检测核酸分子的方法包括：制备含有与待分析的核酸分子特异性杂交的核酸探针的样品溶液和生物样品; 将核酸分子与在制备中制备的样品溶液中的核酸探针缔合; 在关联之后，通过扫描分子计数法计算在准备中制备的样品溶液中包括核酸探针的缔合体的分子数。 用于检测核酸分子的方法还包括：在计算之前从样品溶液中除去蛋白质，或在制备前从生物样品中除去蛋白质。

公开(公告)号：US20130122488A1

公开(公告)日：2013-05-16

申请号：US13746968

申请日：2013-01-22

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Hidetaka Nakata ,  Takuya Hanashi ,  Kunio Hori ,  Kazutaka Nishikawa

IPC: G01N33/53 ,  C12Q1/68

CPC classification number: G01N33/5308 ,  C12Q1/6818 ,  G01N5/045 ,  G01N15/1456 ,  G01N21/6458 ,  G01N2015/1486 ,  G02B21/0048 ,  G02B21/0076

Abstract: There is provided an optical analysis technique enabling the detection of the condition or characteristic of a particle to be observed contained at a low concentration or number density in a sample solution using a light-emitting probe. The inventive optical analysis technique uses an optical system capable of detecting light from a micro region in a solution, such as an optical system of a confocal microscope or a multiphoton microscope, to detect the light from the light-emitting probe having bound to a particle to be observed while moving the position of the micro region in the sample solution (while scanning the inside of the sample solution with the micro region), thereby detecting individually the particle crossing the inside of the micro region to enable the counting of the particle(s) or the acquisition of the information on the concentration or number density of the particle.

Abstract translation: 提供了一种光学分析技术，其能够使用发光探针检测在样品溶液中以低浓度或数量密度包含的待观察颗粒的状态或特性。 本发明的光学分析技术使用能够检测来自诸如共焦显微镜或多光子显微镜的光学系统的解决方案中的微区域的光的光学系统，以检测来自发光探针的光，其结合到颗粒 在移动样品溶液中的微区域的位置（同时用微区域扫描样品溶液的内部）的同时观察，从而单独检测与微区域内部交叉的粒子，以使得能够计数颗粒（ s）或获取关于颗粒的浓度或数量密度的信息。