公开(公告)号：US20180356620A9

公开(公告)日：2018-12-13

申请号：US15002992

申请日：2016-01-21

Applicant: OLYMPUS CORPORATION

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

IPC: G02B21/00

CPC classification number: G02B21/0084 ,  G01N21/6408 ,  G01N21/6458 ,  G02B21/0032 ,  G02B21/0048 ,  G02B21/0076 ,  G02B21/008 ,  G02B21/0088

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：US20180259457A1

公开(公告)日：2018-09-13

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

CPC classification number: G01N21/6428 ,  C12Q1/6816 ,  G01N21/64 ,  G01N21/6408 ,  G01N21/6458 ,  G01N21/78 ,  G01N2021/6439 ,  C12Q2565/101 ,  C12Q2565/601

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：US09528923B2

公开(公告)日：2016-12-27

申请号：US14190780

申请日：2014-02-26

Applicant: OLYMPUS CORPORATION

Inventor： Hidetaka Nakata ,  Tetsuya Tanabe

IPC: G01N15/14 ,  G01N21/64 ,  G01N21/76

CPC classification number: G01N15/14 ,  G01N15/1429 ,  G01N21/6408 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/76 ,  G01N21/763 ,  G01N2021/6419 ,  G01N2021/6421

Abstract: There is provided optical analysis techniques in the scanning molecule counting method using the light measurement with a confocal or multiphoton microscope in which the measuring unit time in the light measurement is set to an appropriate value in order to surely detect an approximately bell shape profile of the signal of a light-emitting particle and avoid excessive increase data volume of time series light intensity data. The inventive optical analysis technique of detecting light of a light-emitting particle in a sample solution generates time series light intensity data of light from a light detection region detected during moving the position of the light detection region of a microscope in the sample solution and detects in the data a signal indicating light from each light-emitting particle individually. The measuring unit time is determined based on the size and the moving speed of the light detection region.

Abstract translation: 在使用共聚焦或多光子显微镜的光测量的扫描分子计数方法中提供了光学分析技术，其中将光测量中的测量单位时间设置为适当的值，以便可靠地检测到大约钟形状 信号发光粒子，避免数据量过大增加时间序列光强数据。 本发明的用于检测样品溶液中的发光粒子的光的光学分析技术产生在移动样品溶液中显微镜的光检测区域的位置时检测到的光检测区域的光的时间序列光强数据，并且检测 在数据中，分别指示来自每个发光颗粒的光的信号。 基于光检测区域的大小和移动速度来确定测量单位时间。

公开(公告)号：US09488578B2

公开(公告)日：2016-11-08

申请号：US14162142

申请日：2014-01-23

Applicant: OLYMPUS CORPORATION

Inventor： Takuya Hanashi ,  Tetsuya Tanabe

IPC: G01N21/64 ,  G01N15/14 ,  G01N21/51 ,  G02B21/00

CPC classification number: G01N21/64 ,  G01N15/14 ,  G01N15/1429 ,  G01N15/1434 ,  G01N21/51 ,  G01N21/6408 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G02B21/0032 ,  G02B21/0076 ,  G02B21/008

Abstract: There is provided a single particle detection technique based on a scanning molecule counting method, enabling individual detection of a single particle using light measurement with a confocal or multiphoton microscope, and quantitative observation of conditions or characteristics of the particle. The inventive technique of detecting a single particle in a sample solution detects light containing substantially constant background light from a light detection region with moving the position of the light detection region of the microscope in a sample solution to generate time series light intensity data; and detects individually a light intensity reduction occurred when a single particle which does not emit light (or a particle whose emitting light intensity in a detected wavelength band is lower than the background light) enters in the light detection region in the time series light intensity data as a signal indicating the existence of each single particle.

Abstract translation: 提供了基于扫描分子计数方法的单粒子检测技术，能够使用共聚焦或多光子显微镜进行光测量来单个检测单个颗粒，并定量观察颗粒的条件或特性。 本发明的检测样品溶液中的单个颗粒的技术通过在样品溶液中移动显微镜的光检测区域的位置来检测来自光检测区域的基本上恒定的背景光的光，以产生时间序列光强度数据; 并且单独检测当不发光的单个颗粒（或检测波长带中的发光强度低于背景光的颗粒）在时间序列光强数据中的光检测区域中时发生的光强度降低 作为指示每个单个颗粒的存在的信号。

公开(公告)号：US09354171B2

公开(公告)日：2016-05-31

申请号：US14162142

申请日：2014-01-23

Applicant: OLYMPUS CORPORATION

Inventor： Takuya Hanashi ,  Tetsuya Tanabe

IPC: G01N21/64 ,  G01N15/14 ,  G01N21/51 ,  G02B21/00

Abstract: There is provided a single particle detection technique based on a scanning molecule counting method, enabling individual detection of a single particle using light measurement with a confocal or multiphoton microscope, and quantitative observation of conditions or characteristics of the particle. The inventive technique of detecting a single particle in a sample solution detects light containing substantially constant background light from a light detection region with moving the position of the light detection region of the microscope in a sample solution to generate time series light intensity data; and detects individually a light intensity reduction occurred when a single particle which does not emit light (or a particle whose emitting light intensity in a detected wavelength band is lower than the background light) enters in the light detection region in the time series light intensity data as a signal indicating the existence of each single particle.

公开(公告)号：US20160139392A1

公开(公告)日：2016-05-19

申请号：US15002992

申请日：2016-01-21

Applicant: OLYMPUS CORPORATION

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

IPC: G02B21/00

CPC classification number: G02B21/0084 ,  G01N21/6408 ,  G01N21/6458 ,  G02B21/0032 ,  G02B21/0048 ,  G02B21/0076 ,  G02B21/008 ,  G02B21/0088

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.

Abstract translation: 提供了一种能够通过扫描分子计数法检测在厚样品中移动的发光物体或发光粒子的显微镜观察技术。 在本发明的技术中，在通过将要观察的区域划分成多个区域而获得的每个观察区域中的光检测区域移动时，检测来自检测到的光检测区域的光的共焦或多光子显微镜的光学系统; 分别检测来自发光粒子的光的信号; 并且在要观察的区域中确定与检测信号相对应的发光粒子的位置。 在每个观察到的子区域中的光检测区域的位置的移动在至少两个方向上连续执行，或者在每个观察到的子区域中连续多次执行。

公开(公告)号：US20140231619A1

公开(公告)日：2014-08-21

申请号：US14266869

申请日：2014-05-01

Applicant: OLYMPUS CORPORATION

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

IPC: G01N15/14

CPC classification number: G01N15/14 ,  G01N15/1429 ,  G01N21/6408 ,  G01N21/6452 ,  G01N21/6458 ,  G01N2015/149 ,  G01N2021/6419 ,  G01N2021/6421 ,  G02B21/00 ,  G02B21/0024 ,  G02B21/0076

Abstract: There is provided a way of enabling the discrimination or identification of the kind of a light-emitting particle corresponding to each pulse form signal in the scanning molecule counting method using the optical measurement by the confocal or multiphoton microscope. In the inventive technique, the position of a light detection region in a sample solution periodically along a predetermined route is moved in measuring the light intensity from the light detection region; and a signal of light from a light-emitting particle is detected individually. Then, an index value indicating a translational diffusional characteristic of one light-emitting particle in a plane perpendicular to the moving direction of the light detection region is determined based upon intensity values of signals of light of the same light-emitting particle for identifying a light-emitting particle.

Abstract translation: 提供了一种使用通过共聚焦或多光子显微镜的光学测量在扫描分子计数方法中能够区分或识别与每个脉冲形式信号相对应的发光粒子的种类的方式。 在本发明的技术中，在从光检测区域测量光强度的同时，移动沿着预定路线周期性地将样品溶液中的光检测区域的位置移动; 并且分别检测来自发光粒子的光的信号。 然后，基于用于识别光的相同发光粒子的光的信号的强度值来确定表示与光检测区域的移动方向垂直的平面中的一个发光粒子的平移扩散特性的指标值 发射粒子

公开(公告)号：US20140175262A1

公开(公告)日：2014-06-26

申请号：US14190780

申请日：2014-02-26

Applicant: OLYMPUS CORPORATION

Inventor： Hidetaka Nakata ,  Tetsuya Tanabe

IPC: G01N15/14

CPC classification number: G01N15/14 ,  G01N15/1429 ,  G01N21/6408 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/76 ,  G01N21/763 ,  G01N2021/6419 ,  G01N2021/6421

Abstract: There is provided optical analysis techniques in the scanning molecule counting method using the light measurement with a confocal or multiphoton microscope in which the measuring unit time in the light measurement is set to an appropriate value in order to surely detect an approximately bell shape profile of the signal of a light-emitting particle and avoid excessive increase data volume of time series light intensity data. The inventive optical analysis technique of detecting light of a light-emitting particle in a sample solution generates time series light intensity data of light from a light detection region detected during moving the position of the light detection region of a microscope in the sample solution and detects in the data a signal indicating light from each light-emitting particle individually. The measuring unit time is determined based on the size and the moving speed of the light detection region.

Abstract translation: 在使用共聚焦或多光子显微镜的光测量的扫描分子计数方法中提供了光学分析技术，其中将光测量中的测量单位时间设置为适当的值，以便可靠地检测到大约钟形状 信号发光粒子，避免数据量过大增加时间序列光强数据。 本发明的用于检测样品溶液中的发光粒子的光的光学分析技术产生在移动样品溶液中显微镜的光检测区域的位置时检测到的光检测区域的光的时间序列光强数据，并且检测 在数据中，分别指示来自每个发光颗粒的光的信号。 基于光检测区域的大小和移动速度来确定测量单位时间。

公开(公告)号：US20140162268A1

公开(公告)日：2014-06-12

申请号：US14178442

申请日：2014-02-12

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/64

CPC classification number: G01N21/64 ,  G01N15/14 ,  G01N15/1429 ,  G01N15/1434 ,  G01N21/51 ,  G01N21/6408 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/6486 ,  G01N2021/6419 ,  G01N2021/6441 ,  G01N2201/103 ,  G01N2201/105 ,  G02B21/0032 ,  G02B21/0076 ,  G02B21/008

Abstract: In the scanning molecule counting method detecting light of a light-emitting particle in a sample solution using a confocal or multiphoton microscope, there is provided an optical analysis technique enabling the scanning in a sample solution with moving a light detection region in a broader area or along a longer route while making the possibility of detecting the same light-emitting particle as different particles as low as possible and remaining the size or shape of the light detection region unchanged as far as possible. In the inventive optical analysis technique, there are performed detecting light from the light detection region and generating time series light intensity data during moving the light detection region along the second route whose position is moved along the first route, and thereby, the signal indicating light from each light-emitting particle in a predetermined route is individually detected using the time series light intensity data.

Abstract translation: 在使用共聚焦或多光子显微镜检测样品溶液中的发光粒子的光的扫描分子计数方法中，提供了一种光学分析技术，其能够在较宽区域中移动光检测区域的样品溶液中进行扫描，或 沿着较长的路线，同时尽可能地检测与不同粒子相同的发光粒子的可能性，并尽可能地保持光检测区域的尺寸或形状不变。 在本发明的光学分析技术中，在沿着沿第一路线移动位置的第二路径移动光检测区域的同时，进行来自光检测区域的检测光并产生时间序列光强度数据，由此，指示光 使用时间序列光强度数据分别检测来自预定路线中的每个发光粒子。

公开(公告)号：US08681332B2

公开(公告)日：2014-03-25

申请号：US13862021

申请日：2013-04-12

Applicant: Olympus Corporation

Inventor： Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/00 ,  G01J1/58

CPC classification number: G01N15/0205 ,  G01N15/1436 ,  G01N21/6458 ,  G02B21/0052 ,  G02B21/0076 ,  G02B21/008

Abstract: There is provided a method of measuring a diffusion characteristic value (for example, a diffusion constant) of a light-emitting particle using the scanning molecule counting method using the optical measurement with a confocal microscope or a multiphoton microscope. The inventive method of measuring a diffusion characteristic value of a light-emitting particle is characterized to measure light intensity from the light detection region with moving the position of the light detection region in the sample solution by changing an optical path of the optical system to generate light intensity data and to compute a diffusion characteristic value of the light-emitting particle based on a deviation time from a moving cycle time of the light detection region in an interval of generation times of two or more signals corresponding to a same light-emitting particle on the light intensity data.

Abstract translation: 提供了一种使用通过使用共聚焦显微镜或多光子显微镜的光学测量的扫描分子计数法来测量发光粒子的扩散特性值（例如，扩散常数）的方法。 本发明的测量发光粒子的扩散特性值的方法的特征在于，通过改变光学系​​统的光路来移动样品溶液中的光检测区域的位置，测量来自光检测区域的光强度，以产生 并且根据与相同的发光粒子的两个以上的信号的产生时间的间隔中的光检测区域的移动周期时间的偏差时间来计算发光粒子的扩散特性值 对光强数据。