Optical Analysis Method for Single Particle Detection Accuracy
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Solution Overview
Problem
Current optical analysis techniques using confocal microscopes struggle to accurately detect and quantify light-emitting particles at low concentrations due to overlapping signals from multiple particles, leading to inaccurate counting and concentration measurements.
Innovation Solution
An optical analysis method that adjusts the apparent light detection region by setting a threshold value for signal detection, ensuring only single particles are detected, even at high concentrations, by modifying the analysis processing to define a narrower detection region based on light intensity distribution and moving the light detection region within the sample solution.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Area of stationary object
If the light detection region is made larger to detect more particles, then the detection coverage is improved, but multiple particles overlap causing signal confusion and reducing measurement precision
Solution Approach 1:
The patent dynamically adjusts the threshold parameter for signal detection based on the concentration of particles in the sample. By changing this parameter, the system can adapt to different particle densities, effectively narrowing the detection region's impact when particle concentration is high, thus preventing signal overlap while maintaining detection sensitivity
Solution Approach 2:
The system dynamically adjusts the light detection region size and threshold values based on real-time particle concentration measurements. This dynamic adaptation allows the detection region to effectively shrink or expand, preventing particle signal overlap at high concentrations while maintaining comprehensive detection at low concentrations
2Measurement precision
If the threshold value is lowered to detect fainter signals, then detection sensitivity is improved, but noise increases leading to false detections
Solution Approach 1:
The system uses feedback from the detected signal distribution and concentration measurements to dynamically adjust the threshold value. By continuously monitoring the signal characteristics and adapting the threshold accordingly, the system maintains high detection sensitivity while filtering out noise, preventing both false negatives and false positives
Solution Approach 2:
The threshold parameter is dynamically changed based on the measured particle concentration and signal distribution characteristics. This parameter adaptation allows the system to optimize detection sensitivity for faint signals while maintaining reliability by raising the threshold when noise levels increase
3Measurement precision
If statistical processing is applied to analyze fluorescence intensity fluctuations, then molecular characteristics can be determined, but the concentration range is limited and measurement time increases
Solution Approach 1:
The patent extracts individual particle signals from the overall fluorescence intensity data by applying dynamic thresholding and signal segmentation. This extraction method isolates single-particle events from the statistical fluctuations, enabling direct counting and concentration measurement without requiring lengthy autocorrelation analysis, thus reducing measurement time while maintaining precision
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 method enhances the accuracy of detecting and counting light-emitting particles, expanding the concentration range for reliable measurements and improving the precision of particle concentration and number density determination.
Implementation Method 1
the light emitted from a light-emitting particle may be fluorescence
Implementation Method 2
detection and/or measurement of faint light at a single photon or single fluorescent molecule level have become possible by using an optical system of a confocal microscope and a super high sensitive light detection technique capable of the photon counting
Data Source
AI summary
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.