Olympus Corporation

Takuya Hanashi

Tetsuya Tanabe

Mitsushiro Yamaguchi

Hidetaka Nakata

Kazutaka Nishikawa

Kunio Hori

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.

Optical analysis method and optical analysis device using single light-emitting particle detection

This method for detecting a target particle comprises (a) preparing a solution containing a target particle, a luminescent probe that binds to the target particle and a particle for separation and recovery, or containing the target particle bound to the luminescent probe, the luminescent probe and the particle for separation and recovery, and forming a complex composed of the target particle, the luminescent probe and the particle for separation and recovery in the solution, (b) recovering the particle for separation and recovery from the solution by solid-liquid separation treatment after the (a) and preparing a sample solution containing the particle for separation and recovery, and (c) calculating the number of the complex present in the sample solution according to a scanning molecule counting method, wherein the particles for separation and recovery bind to a complex composed of the target particles and the luminescent probe.

Method for detecting target particle

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.

Method for detecting a target particle

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.

Single particle detection device, single particle detection method, and computer program for single particle detection, using optical analysis

There is provided a structure to make the setting of a criterion for eliminating noises easy in the scanning molecule counting method. 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 in the sample solution is generated, and a signal of a light-emitting particle individually is detected in the time series light intensity data, wherein a signal having a light intensity in a light intensity range set based upon a signal generation frequency integrated value distribution which is a distribution, obtained by using as a variable an intensity of a signal, of integrated values of generation frequencies of signals having an intensity not lower than the variable is extracted as the signal of the light-emitting particle.

Optical analysis device, optical analysis method and computer program for optical analysis using single light-emitting particle detection

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.

Method of detecting sparse particles in a solution using a light-emitting probe

There is provided a single particle detection technique based on the scanning molecule counting method which individually detects single particles using light measurement with a confocal or multiphoton microscope, where the existences of a non-light-emitting particle and a light-emitting particle can be detected while being discriminated from one another in a sample solution. The inventive technique of detecting a single particle detects light from a light detection region during moving the position of the light detection region of the microscope in a sample solution containing a non-light-emitting particle and a light-emitting particle to generate time series light intensity data; and detects in the time series light intensity data a light intensity increase relative to background light intensity as a signal indicating the existence of the light-emitting particle and a light intensity reduction relative to the background light intensity as a signal indicating the existence of the non-light-emitting particle.

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.

Optical analysis device and optical analysis method using a wavelength characteristic of light of a single light-emitting particle

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.

Growing community of inventors

Hachioji, Japan

Takuya Hanashi