Flow Matrix Fluorescence Analysis With Ellipse Gating
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Solution Overview
Problem
Current flow matrix analyzers face challenges in quantitative analysis due to cumbersome steps, long analysis times, human subjectivity in threshold setting, and fluorescence wavelength interference, leading to inaccurate results.
Innovation Solution
A method and apparatus for analyzing flow matrix data that includes fluorescence compensation, ellipse gate construction, and automatic classification of microsphere particles, using FITC, PE, and PerCP fluoresceins, to enhance accuracy and simplify the analysis process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If manual threshold setting is used during analysis, then the analysis process is simple to operate, but the quantitative accuracy is affected due to human subjectivity
Solution Approach 1:
The system performs automatic threshold determination through image processing algorithms that analyze the binary image data and autonomously establish classification thresholds, eliminating the need for manual intervention and removing human subjectivity from the quantification process
Solution Approach 2:
The manual mechanical process of setting thresholds by human operators is replaced with automated computational image processing algorithms that objectively determine thresholds based on data analysis, substituting human judgment with algorithmic decision-making
2Device complexity
If fixed position threshold is used in each measurement, then the analysis steps are simplified, but the quantitative accuracy deteriorates due to poor quantitative accuracy
Solution Approach 1:
The threshold determination transitions from a static fixed position approach to a dynamic adaptive process where thresholds are automatically adjusted based on the actual distribution characteristics of the measured data, allowing the system to adapt to different measurement conditions while maintaining simplicity
3Productivity
If fluorescence compensation is not performed, then the analysis process is faster and simpler, but the accuracy of detection results deteriorates due to mutual interference of fluorescences
Solution Approach 1:
Fluorescence compensation is performed as a preliminary processing step before the main classification and quantification processes, correcting the spectral overlap issues in advance so that subsequent analysis can proceed quickly without sacrificing accuracy
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
The method and apparatus provide high-accuracy quantitative and qualitative analysis by automating the classification process, reducing interference, and improving the precision of concentration calculations.
Implementation Method 1
The flow matrix analyzer measures various biophysical properties of particles by detecting fluorescence pulse signals specifically labeled with microsphere particles specificity
Implementation Method 2
acquiring and sampling a forward-scattered light pulse signal outputted by a flow matrix analyzer
Data Source
AI summary
A method for analyzing flow matrix data includes: acquiring and sampling a forward-scattered light pulse signal outputted by a flow matrix analyzer and a plurality of fluorescence pulse signals; performing fluorescence compensation on three pieces of fluorescence pulse data after the sampling, and mapping true data of two classification fluorescence pulses to a scatter plot so as to form a two-dimensional array; converting the two-dimensional array into a binary image, and constructing an ellipse gate related to the gathering region; obtaining intrinsic characteristics and movement characteristics of the ellipse gate; and obtaining coordinates of two focuses on the ellipse gate; calculating a distance between each microsphere particle and the two focuses on each ellipse gate; and calculating a median value of true data of calibration fluorescence pulses on all microsphere particles.