公开(公告)号：US20140134608A1

公开(公告)日：2014-05-15

申请号：US14162142

申请日：2014-01-23

Applicant: Olympus Corporation

Inventor： Takuya Hanashi ,  Tetsuya Tanabe

IPC: G01N21/51 ,  G01N21/64

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：US09863806B2

公开(公告)日：2018-01-09

申请号：US13946091

申请日：2013-07-19

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe

IPC: G01J1/42 ,  G01N21/64 ,  G02B21/00 ,  G02B21/16

CPC classification number: G01J1/42 ,  G01N21/6452 ,  G02B21/0076 ,  G02B21/16

Abstract: In the scanning molecule counting method using the light measurement with a confocal microscope or a multiphoton microscope, a measuring time is optimized with suppressing the scattering in a result small irrespective of light-emitting particle concentrations. In the inventive technique of detecting and analyzing the light from an light-emitting particle, there are repeated processes of detecting the light intensity from a light detection region with moving the position of the light detection region of an optical system in a sample solution by changing the optical path of the optical system of the microscope, and detecting the signals of the light of light-emitting particles individually, and based on the time taken for the number of the signals from the light-emitting particles to reach a predetermined number, the light-emitting particle concentration in the sample solution is determined.

公开(公告)号：US09423349B2

公开(公告)日：2016-08-23

申请号：US14178442

申请日：2014-02-12

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Mitsushiro Yamaguchi

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

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：US09329117B2

公开(公告)日：2016-05-03

申请号：US14266869

申请日：2014-05-01

Applicant: OLYMPUS CORPORATION

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

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

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：US09103718B2

公开(公告)日：2015-08-11

申请号：US13888868

申请日：2013-05-07

Applicant: OLYMPUS CORPORATION

Inventor： Takuya Hanashi ,  Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/00 ,  G01N21/17 ,  G01J1/16 ,  G01N21/64 ,  G01N15/14 ,  G02B21/00 ,  G01J1/58 ,  G01C21/02 ,  G02B21/26

CPC classification number: G01J1/16 ,  G01C21/02 ,  G01J1/58 ,  G01N15/1463 ,  G01N21/6452 ,  G01N21/6458 ,  G01N2021/6421 ,  G02B21/0032 ,  G02B21/0076 ,  G02B21/26

Abstract: The inventive technique of detecting and analyzing light from a light-emitting particle in accordance with the scanning molecule counting method using an optical measurement with a confocal microscope or a multiphoton microscope is characterized by detecting intensities of components of two or more wavelength bands of light from a light detection region of an optical system with moving the position of the light detection region in a sample solution by changing the optical path of the optical system of the microscope; detecting individually signals of the light from each light-emitting particle in the intensities of the components of the two or more wavelength bands of the detected light; and identifying a kind of light-emitting particle based on the intensities of the components of the two or more wavelength bands of the signals of the light of the detected light-emitting particle.

Abstract translation: 根据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检测和分析来自发光粒子的光的本发明的技术的特征在于检测两个或更多个波长带的光的分量的强度， 光学系统的光检测区域，通过改变显微镜的光学系统的光路来移动样品溶液中的光检测区域的位置; 在检测到的光的两个或更多个波长带的分量的强度中单独地检测来自每个发光粒子的光的信号; 并且基于检测到的发光粒子的光的信号的两个或更多波长带的分量的强度来识别发光粒子的种类。

公开(公告)号：US08803106B2

公开(公告)日：2014-08-12

申请号：US13864867

申请日：2013-04-17

Applicant: Olympus Corporation

Inventor： Mitsushiro Yamaguchi ,  Tetsuya Tanabe

IPC: G01J1/58 ,  G01N21/64 ,  G02B21/00

CPC classification number: G01N21/6445 ,  G01N15/1434 ,  G01N15/1456 ,  G01N21/6408 ,  G01N21/6458 ,  G02B21/0068 ,  G02B21/0076 ,  G02B21/0092

Abstract: There is provided an optical analysis technique which observes a polarization characteristic of a light-emitting particle using the scanning molecule counting method using an optical measurement with a confocal microscope or a multiphoton microscope. In the inventive optical analysis technique, the light detection region is irradiated with excitation light consisting of predetermined polarized light component(s) and the intensity of at least one polarized light component of the light from the light detection region is detected with moving the position of the light detection region of the optical system in a sample solution; a signal of each light-emitting particle is detected individually in the intensity of at least one polarized light component; and based on the intensity of at least one polarized light component of the signal of the detected light-emitting particle, the polarization characteristic value of the light-emitting particle is computed.

Abstract translation: 提供了一种光学分析技术，其使用通过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方法观察发光粒子的偏振特性。 在本发明的光学分析技术中，用由预定偏振光分量组成的激发光照射光检测区域，并且通过移动光检测区域的位置来检测来自光检测区域的光的至少一个偏振光分量的强度 样品溶液中的光学系统的光检测区域; 以至少一个偏振光分量的强度分别检测每个发光粒子的信号; 并且基于检测到的发光粒子的信号的至少一个偏振光分量的强度，计算发光粒子的偏振特性值。

公开(公告)号：US20140170760A1

公开(公告)日：2014-06-19

申请号：US14188375

申请日：2014-02-24

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N21/64

CPC classification number: G01N21/64 ,  G01N21/6408 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N21/6486 ,  G02B21/0032 ,  G02B21/0048 ,  G02B21/0076 ,  Y10T436/143333

Abstract: There is provided an optical analysis technique of detecting light of a light-emitting particle in a sample solution in the scanning molecule counting method using the light measurement with a confocal or multiphoton microscope, for suppressing the scattering in detected results of signals of light of light-emitting particles smaller and achieving the improvement of accuracy. The inventive technique comprises moving the position of a light detection region along a predetermined route for multiple circulation times by changing the optical path of the optical system; detecting light from the light detection region and generating time series light intensity data during the moving of the light detection region and detecting individually a signal indicating light from each light-emitting particle existing in the predetermined route using the time series light intensity data obtained in the circulating movements of the light detection region of multiple times.

Abstract translation: 提供了一种使用共聚焦或多光子显微镜的光测量的扫描分子计数法中的样品溶液中的发光粒子的光的光学分析技术，用于抑制光的信号的检测结果中的散射 - 更小的颗粒，实现精度的提高。 本发明的技术包括通过改变光学系​​统的光路来沿着预定路线移动光检测区域的位置以进行多次循环; 检测来自光检测区域的光，并且在光检测区域移动期间产生时间序列光强度数据，并且使用在所述光检测区域中获得的时间序列光强度数据，分别检测指示存在于预定路线中的每个发光粒子的光的信号 光检测区域的循环运动多次。

公开(公告)号：US20140131593A1

公开(公告)日：2014-05-15

申请号：US14158279

申请日：2014-01-17

Applicant: OLYMPUS CORPORATION

Inventor： Hidetaka Nakata ,  Tetsuya Tanabe

IPC: G01N21/64

CPC classification number: G01N21/6408 ,  G01N21/6428 ,  G01N21/645 ,  G01N21/6452 ,  G01N21/6458 ,  G01N2021/6419 ,  G01N2021/6421 ,  G01N2021/6439 ,  G02B21/0076

Abstract: A method for detecting a fluorescent particle comprises the preparation of a sample solution containing fluorescent particles and a substance that promotes transition of the fluorescent particles from a triplet excited state to a singlet ground state, and calculation of the number of molecules of fluorescent particles present in the prepared sample solution. Calculation of the number of molecules of the fluorescent particles comprises moving the location of a photodetection region of an optical system in the sample solution using the optical system of a confocal microscope or multi-photon microscope, individually detecting fluorescent particles by detecting a light signal from the fluorescent particles present in the photodetection region while moving the location of the photodetection region in the sample solution, and counting the number of fluorescent particles detected during movement of the location of the photodetection region by counting the number of individually detected fluorescent particles.

Abstract translation: 检测荧光粒子的方法包括制备含有荧光粒子的样品溶液和促进荧光粒子从三线态激发态转变为单线态的物质，并计算存在于荧光粒子中的荧光粒子的分子数 制备的样品溶液。 荧光粒子的分子数的计算包括使用共焦显微镜或多光子显微镜的光学系统移动样品溶液中的光学系统的光电检测区域的位置，通过检测荧光颗粒，通过检测来自 存在于光检测区域中的荧光体，同时移动样品溶液中的光电检测区域的位置，并且通过计数单独检测到的荧光颗粒的数量来计数在光检测区域的位置移动期间检测的荧光颗粒的数量。

公开(公告)号：US20140024020A1

公开(公告)日：2014-01-23

申请号：US14036921

申请日：2013-09-25

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe

IPC: G01N21/64

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：US20130314705A1

公开(公告)日：2013-11-28

申请号：US13862021

申请日：2013-04-12

Applicant: OLYMPUS CORPORATION

Inventor： Tetsuya Tanabe ,  Mitsushiro Yamaguchi

IPC: G01N15/02

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。