公开(公告)号：US09494779B2

公开(公告)日：2016-11-15

申请号：US14451021

申请日：2014-08-04

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe

IPC: G02B21/00 ,  G01N21/64 ,  G01N15/14 ,  G02B21/16 ,  G01N21/84

CPC classification number: G02B21/0084 ,  G01N15/1429 ,  G01N15/1434 ,  G01N21/6408 ,  G01N21/6458 ,  G01N21/6486 ,  G01N2021/8405 ,  G01N2201/12 ,  G02B21/0004 ,  G02B21/0076 ,  G02B21/16

Abstract: In the optical analysis technique of detecting an existence of a single particle in a sample solution with a confocal microscope or a multiphoton microscope according to the scanning molecule counting method of the present invention, the position of a light detection region is moved in the sample solution; the light intensity from the light detection region is measured so that light intensity data will be generated; a first occurrence probability in assuming a first condition that no single particles exist in the light detection region and a second occurrence probability in assuming a second condition that a single particle exists in the light detection region for a time variation of light intensity value on the light intensity data are computed; and a signal indicating each single particle is detected based on those occurrence probabilities, and thereby enabling improvements in the sensitivity and/or S/N ratio.

Abstract translation: 在根据本发明的扫描分子计数法的共聚焦显微镜或多光子显微镜检测样品溶液中的单一颗粒的存在的光学分析技术中，光检测区域的位置在样品溶液中移动 ; 测量来自光检测区域的光强度，以便产生光强度数据; 假定在光检测区域中不存在单个颗粒的第一条件的第一个发生概率，以及假设在光检测区域中存在单个颗粒存在于光的光强度值的时间变化的第二条件的第二个出现概率 计算强度数据; 并且基于那些发生概率检测表示每个单个粒子的信号，从而能够提高灵敏度和/或S / N比。

公开(公告)号：US08958066B2

公开(公告)日：2015-02-17

申请号：US13789111

申请日：2013-03-07

Applicant: Olympus Corporation

Inventor： Takuya Hanashi ,  Tetsuya Tanabe ,  Mitsushiro Yamaguchi ,  Hidetaka Nakata

IPC: G01N21/00 ,  G01N21/64

CPC classification number: G01N21/6486 ,  G01N21/6428 ,  G01N21/6452 ,  G01N21/6458

Abstract: There is provided an optical analysis technique enabling identification of a kind of light-emitting particle corresponding to a signal on a time series light intensity data or identification of a signal corresponding to light-emitting particles other than a particle to be observed in an optical measurement using a confocal microscope or a multiphoton microscope. The inventive optical analysis technique measures simultaneously and separately intensities of lights of two or more wavelength bands from a light detection region in a sample solution containing light-emitting particles of two or more kinds to generate time series light intensity data of the respective wavelength bands; detects signals simultaneously generated on the time series light intensity data of at least two wavelength bands; and identifies the simultaneously generated signals as signals of a light-emitting particle of at least one specific kind.

Abstract translation: 提供了一种光学分析技术，其能够对与光学测量中要观察的颗粒以外的发光粒子相对应的时间序列光强度数据或对应于发光粒子的信号的识别相对应的一种发光粒子 使用共焦显微镜或多光子显微镜。 本发明的光学分析技术测量来自含有两种或更多种发光粒子的样品溶液中的光检测区域的两个或更多个波长带的光的同时且分开的强度，以产生各波长带的时间序列光强度数据; 检测同时产生的至少两个波长带的时间序列光强数据的信号; 并将同时生成的信号识别为至少一种特定种类的发光粒子的信号。

公开(公告)号：US08680485B2

公开(公告)日：2014-03-25

申请号：US13840122

申请日：2013-03-15

Applicant: Olympus Corporation

Inventor： Tetsuya Tanabe

IPC: G01N21/64 ,  G01J1/58

CPC classification number: G01N21/85 ,  G01N15/1434 ,  G01N21/6452 ,  G01N21/6458 ,  G01N2021/6419 ,  G01N2021/6421 ,  G01N2201/1087 ,  G02B21/0052 ,  G02B21/0076 ,  G02B21/008

Abstract: There is provided a method of avoiding deterioration of the accuracy in the number of detected light-emitting particles due to that two or more light-emitting particles are encompassed at a time in the light detection region in the scanning molecule counting method using an optical measurement with a confocal microscope or a multiphoton microscope. In the inventive optical analysis technique, in the detection of an individual signal indicating light of a light-emitting particle by selectively detecting a signal having an intensity beyond a threshold value as a signal indicating light of a light-emitting particle in light intensity data produced through measuring light intensity during moving the position of a light detection region in a sample solution, the threshold value is set so that a signal indicating light from a light-emitting particle encompassed in a region narrower than the light detection region will be detected selectively.

Abstract translation: 提供了一种避免由于在使用光学测量的扫描分子计数方法中在光检测区域中一次包围两个或更多个发光粒子而使检测到的发光粒子数量的精度降低的方法 用共焦显微镜或多光子显微镜。 在本发明的光学分析技术中，在通过选择性地检测具有超过阈值的强度的信号来检测指示发光粒子的光的单个信号作为产生光强度数据中的发光粒子的光的信号 通过在移动样品溶液中的光检测区域的位置时测量光强度，设定阈值，以便选择性地检测到包含在比光检测区域窄的区域中的发光粒子的光的信号。

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：US11016026B2

公开(公告)日：2021-05-25

申请号：US15983412

申请日：2018-05-18

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Takuya Hanashi ,  Hidetaka Nakata

IPC: G01N21/64 ,  C12Q1/6816 ,  G01N21/78

Abstract: In the scanning molecule counting method of measuring light intensity from a light detection region while moving the position of the light detection region of a confocal or multiphoton microscope in a sample solution containing light-emitting particles, generating time series light intensity data and detecting each of signals of the light-emitting particles individually in the data, wherein the light-emitting particles are formed by binding to a particle to be observed a light-emitting probe which emits light through binding to the particle to be observed and in which a stochastic transition between a non-light-emitting state and a light-emitting state occurs in the unbound state, the moving speed of the position of the light detection region is adjusted to make the time during which the unbound probe is encompassed by the moving light detection region longer than an average lifetime during which the probe is in the light-emitting state.

公开(公告)号：US10310245B2

公开(公告)日：2019-06-04

申请号：US15002992

申请日：2016-01-21

Applicant: OLYMPUS CORPORATION

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

IPC: G02B21/00 ,  G01N21/64

Abstract: There is provided a microscopic observation technique capable of detecting a light-emitting object or a light-emitting particle moving in a thick sample by the scanning molecule counting method. In the inventive technique, the light from a light detection region of is detected the optical system of a confocal or multiphoton microscope is detected with while moving the light detection region in each observed subregion obtained by dividing a region to be observed into plural regions; the signal of the light from a light-emitting particle is individually detected; and the position of the light-emitting particle corresponding to the detected signal is determined in the region to be observed. The moving of the position of the light detection region in each observed subregion is performed continuously in at least two directions or and/or continuously multiple times in each observed subregion.

公开(公告)号：US09395357B2

公开(公告)日：2016-07-19

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: G01N21/76 ,  G01N33/53 ,  G01N5/04 ,  G01N15/14 ,  G01N21/64 ,  C12Q1/68 ,  G02B21/00

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。

公开(公告)号：US09068944B2

公开(公告)日：2015-06-30

申请号：US14036921

申请日：2013-09-25

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe

IPC: G01N21/64 ,  G01N15/14 ,  G02B21/00 ,  G02B21/16 ,  G02B21/36 ,  G01N21/51 ,  G01N21/76

CPC classification number: G01N21/64 ,  G01N15/14 ,  G01N15/1429 ,  G01N15/1434 ,  G01N21/51 ,  G01N21/6408 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/76 ,  G01N21/763 ,  G01N2015/0053 ,  G01N2015/1006 ,  G02B21/0076 ,  G02B21/16 ,  G02B21/367

Abstract: There is provided a structure to reduce the size of light intensity data in the scanning molecule counting method using an optical measurement with a confocal microscope or a multiphoton microscope. In the inventive optical analysis technique of detecting light of a light-emitting particle in a sample solution, time series light intensity data of light from a light detection region detected with moving the position of the light detection region of the microscope in the sample solution is generated, and a signal of a light-emitting particle individually is detected in the time series light intensity data. In that case, regions where no signal indicating light of light-emitting particles exist in the time series light intensity data is removed from the time series light intensity data.

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在扫描分子计数方法中减小光强度数据的尺寸的结构。 在本发明的检测样品溶液中的发光粒子的光的光学分析技术中，通过移动样品溶液中的显微镜的光检测区域的位置而检测到的光检测区域的光的时间序列的光强度数据是 并且在时间序列光强度数据中检测发光粒子的信号。 在这种情况下，从时间序列光强度数据中除去在时间序列光强度数据中不存在表示发光粒子的光的信号的区域。

公开(公告)号：US20140339444A1

公开(公告)日：2014-11-20

申请号：US14451021

申请日：2014-08-04

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe

IPC: G02B21/00 ,  G01N21/64

CPC classification number: G02B21/0084 ,  G01N15/1429 ,  G01N15/1434 ,  G01N21/6408 ,  G01N21/6458 ,  G01N21/6486 ,  G01N2021/8405 ,  G01N2201/12 ,  G02B21/0004 ,  G02B21/0076 ,  G02B21/16

Abstract: In the optical analysis technique of detecting an existence of a single particle is a sample solution with a confocal microscope or a multiphoton microscope according to the scanning molecule counting method of the present invention, the position of a light detection region is moved in the sample solution; the light intensity from the light detection region is measured so that light intensity data will be generated; a first occurrence probability in assuming a first condition that no single particles exist in the light detection region and a second occurrence probability in assuming a second condition that a single particle exists in the light detection region for a time variation of light intensity value on the light intensity data are computed; and a signal indicating each single particle is detected based on those occurrence probabilities, and thereby enabling improvements in the sensitivity and/or S/N ratio.

Abstract translation: 在通过本发明的扫描分子计数法的共聚焦显微镜或多光子显微镜的检测单粒子的存在的光学分析技术中，光检测区域的位置在样品溶液中移动 ; 测量来自光检测区域的光强度，以便产生光强度数据; 假定在光检测区域中不存在单个颗粒的第一条件的第一个发生概率，以及假设在光检测区域中存在单个颗粒存在于光的光强度值的时间变化的第二条件的第二个出现概率 计算强度数据; 并且基于那些发生概率检测表示每个单个粒子的信号，从而能够提高灵敏度和/或S / N比。

公开(公告)号：US08785886B2

公开(公告)日：2014-07-22

申请号：US13788972

申请日：2013-03-07

Applicant: Olympus Corporation

Inventor： Kazutaka Nishikawa ,  Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/64 ,  H01J65/06 ,  G01N15/14

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。