Optical Analysis Device Using Scanning Light Detection Region for Particle Identification
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Solution Overview
Problem
The scanning molecule counting method faces challenges in distinguishing and identifying light-emitting particles with common emission wavelengths due to varying brightness, as the absolute light intensity measured is not an inherent property of the particle and is affected by the position within the light detection region, making it difficult to differentiate between particles with different brightness levels.
Innovation Solution
An optical analysis device and method that detects light from light-emitting particles using a confocal or multiphoton microscope, where the light detection region is moved periodically to capture light intensity changes over multiple cycles, allowing for the determination of a translational diffusional characteristic index value based on intensity variations, enabling discrimination and identification of particles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the scanning molecule counting method is used to detect light from light-emitting particles, then the concentration and number density of particles can be detected, but it becomes difficult to distinguish and identify particles with common emission wavelengths due to varying brightness
Solution Approach 1:
The invention changes the parameter used for particle identification from absolute brightness (light intensity) to relative brightness ratio. By comparing the light intensity of a particle at different time points during the scanning process, the system derives a ratio that characterizes each particle's brightness relative to the excitation light intensity at different positions, enabling identification of particles with common emission wavelengths but different brightness characteristics
Solution Approach 2:
The invention replaces the conventional approach of using absolute light intensity measurements with a ratio-based calculation method. Instead of relying on the absolute brightness value which varies with particle position in the excitation light field, the system uses the ratio of light intensities at different time points, substituting a mathematical transformation to eliminate position-dependent variations
2Productivity
If particles with different brightness levels are measured using the scanning molecule counting method, then light intensity data can be collected, but the absolute light intensity is not an inherent property of the particle and is affected by position within the light detection region
Solution Approach 1:
The invention transforms the unreliable absolute light intensity parameter into a reliable relative brightness ratio parameter. By calculating the ratio of light intensities at different time points during scanning, the system eliminates the influence of particle position within the excitation light field, making the brightness measurement an inherent property of the particle rather than dependent on measurement conditions
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the effective discrimination and identification of light-emitting particles, even at lower concentrations, without the need for particles with different emission wavelengths, enhancing the ability to detect and quantify particles in complex samples.
Implementation Method 1
the light emitted from a light-emitting particle may be fluorescence
Implementation Method 2
the light emitted from a light-emitting particle may be fluorescence, phosphorescence, chemiluminescence, bioluminescence, scattered light
Implementation Method 3
a light-emitting particle being dispersed and moving at random in the sample solution
Data Source
AI summary
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.