Autofluorescence Spectrum Correction for Particle Population Separation
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Solution Overview
Problem
Existing flow cytometry methods struggle to accurately analyze autofluorescence from microparticles due to varying autofluorescence levels among different populations, making it difficult to separate and correct for autofluorescence to enhance analysis accuracy.
Innovation Solution
A particle analysis system and method that includes a light detector and information processing unit to acquire and process autofluorescence data, creating spectral plots to identify and record autofluorescence populations, facilitating their separation and correction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If autofluorescence correction is performed using conventional flow cytometry methods, then analysis accuracy can be improved, but it becomes difficult to separate and identify multiple autofluorescence populations with different autofluorescence characteristics
Solution Approach 1:
The patent transitions from conventional two-dimensional flow cytometry plots to three-dimensional spectral plots that incorporate wavelength information as a third dimension. This dimensional expansion enables the separation and identification of multiple autofluorescence populations by their distinct spectral signatures, resolving the difficulty of population separation while maintaining measurement precision
Solution Approach 2:
The patent introduces spectral reference data as an intermediary tool that mediates between the complex autofluorescence signals and the analysis process. By comparing measured spectra against reference spectra of known autofluorescence sources, the system can identify and separate different autofluorescence populations, improving both detection capability and analysis accuracy
2Loss of information
If spectral plots with logarithmic axis are used to display low intensity data, then the spectrum of low intensity microparticles can be visualized, but the spectrum appears with excessively high dispersion and negative numbers cannot be displayed
Solution Approach 1:
The patent implements dynamic axis scaling in spectral plots where the display range and scaling are automatically adjusted based on the intensity range of the data being visualized. This dynamic adaptation allows low intensity spectra to be displayed with appropriate resolution without the excessive dispersion problems of fixed logarithmic scales, while maintaining the ability to represent the full dynamic range of intensities
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
Enables accurate identification and separation of multiple autofluorescence populations, improving the analysis of microparticles by correcting for autofluorescence variations.
Implementation Method 1
measures the intensity and spectrum of fluorescence or scattered light emitted from the microparticles
Implementation Method 2
measures the intensity and spectrum of fluorescence or scattered light emitted from the microparticles
Data Source
AI summary
A particle analysis system comprising: a light detector that acquires light generated by irradiating a particle with excitation light; and an information processing unit that outputs a spectral plot including spectrum information of an autofluorescence population specified in a two-dimensional plot of measurement data each of which corresponds to the acquired light and spectrum information of the measurement data and that records the spectrum information of the autofluorescence population as an autofluorescence reference spectrum in a fluorescence separation process.