Chromatography Fluorescence Analysis for Overlapping Multi-Dye Signals
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Solution Overview
Problem
Existing analysis methods struggle to accurately identify fluorescence emissions from multiple fluorescent substances due to spectral and space-time overlaps, particularly when using RGB color sensors that can detect only three colors, making it difficult to analyze components labeled with four or more fluorescent substances.
Innovation Solution
An analyzer is configured to separate samples by chromatography, acquire time-series fluorescence data in multiple wavelength bands, and compare this data with stored model fluorescence signals to determine the specific fluorescent substances labeling the components, even when there is a spectral and space-time overlap.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If N-color detection (N=3 for RGB sensor) is used to detect fluorescence from M kinds of fluorescent substances, then device complexity is reduced and cost is decreased, but measurement precision deteriorates when M>N due to spectral and space-time overlaps
Solution Approach 1:
The patent stores model fluorescence signals for each fluorescent substance in advance in a database. During analysis, these pre-stored models are compared with actual detection signals to identify fluorescent substances. This preliminary preparation of reference data enables accurate identification even when M>N, resolving the contradiction between using simple RGB sensors and achieving precise multi-fluorescent substance detection.
Solution Approach 2:
The patent creates and stores copies of characteristic fluorescence signals (model signals) for each fluorescent substance under various conditions (with/without other substances present). These copied reference signals are then used for comparison with actual detection signals, enabling identification without requiring complex real-time spectral decomposition, thus allowing accurate detection with simple RGB sensors.
2Adaptability or versatility
If M kinds of fluorescent substances are used to label components, then analysis versatility is improved, but difficulty of detecting and measuring increases due to spectral and space-time overlaps when M>N
Solution Approach 1:
The patent stores copies of model fluorescence signals for each fluorescent substance in a database, including signals obtained when each substance is present alone and when present with other substances. During detection, these stored models are compared with actual signals to identify fluorescent substances, enabling versatile multi-component analysis without increased detection difficulty.
Solution Approach 2:
The patent compares fluorescence signals across multiple wavelength bands (R, G, B) and time points to identify fluorescent substances. By analyzing parameter variations (wavelength distribution and temporal characteristics) rather than relying on single-point measurements, the system achieves accurate identification even with spectral and space-time overlaps.
3Manufacturing precision
If spectral and space-time overlaps are present among fluorescent substances, then manufacturing precision of labeling is improved, but loss of information increases due to inability to distinguish individual emissions
Solution Approach 1:
The patent pre-stores model fluorescence signals that represent the characteristic emission patterns of each fluorescent substance, including how they appear with and without other substances present. This preliminary data preparation preserves information about individual substance emissions, enabling later recovery and identification even when overlaps occur during actual detection.
Solution Approach 2:
The patent compares actual detection signals with stored model signals and uses this comparison feedback to identify which fluorescent substances are present. By continuously referencing the stored models and adjusting identification based on match quality, the system recovers lost information about individual emissions despite spectral and space-time overlaps.
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 detection of multiple components labeled with M kinds of fluorescent substances, even when M>N, by accurately identifying which substances are present despite spectral and space-time overlaps.
Implementation Method 1
separating a sample including a plurality of components labeled with any of M kinds of fluorescent substances by chromatography
Implementation Method 2
a plurality of kinds of components included in samples including biological samples, such as DNA, proteins, and cells, is labeled with a plurality of kinds of fluorescent substances (where the correspondence is not always one-to-one correspondence), fluorescence emitted from the plurality of kinds of fluorescent substances is detected being identified
Data Source
AI summary
An analysis system includes an analyzer configured to separate a sample including a plurality of components labeled with any of M kinds of fluorescent substances by chromatography and acquire first time-series data of fluorescence signals detected in N kinds (M>N) of wavelength bands in a state in which at least a part of the plurality of components is not completely separated; and a computer configured to compare the first time-series data with the second time-series data, and determine which kind of fluorescent substance of M kinds of fluorescent substances individually labels each of the plurality of components.