Capillary Electrophoresis Noise Removal via Multi-Wavelength Matrix Conversion
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Solution Overview
Problem
In DNA sequencing using capillary electrophoresis, noise fluorescence peaks from impurities can overlay or be detected separately, leading to inaccurate determination of base species due to their distinct fluorescence spectra, which are not properly accounted for in conventional matrix conversion methods.
Innovation Solution
The method involves determining the fluorescence intensity ratio for noise peaks in multiple wavelength bands and performing a matrix conversion that includes both labeled and noise fluorescent substances to isolate and remove noise peaks, ensuring accurate intensity calculation for labeled fluorescent substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional matrix conversion methods are used to analyze fluorescence spectra, then the analysis process is simple, but noise fluorescence peaks from impurities are not properly accounted for, leading to inaccurate determination of base species
Solution Approach 1:
The fluorescence spectrum is segmented into multiple wavelength bands, and noise peaks are identified and separated from valid signal peaks. The matrix conversion process is divided into handling noise peaks and valid peaks separately, allowing accurate determination of base species while maintaining manageable process complexity through systematic segmentation of the analysis task.
Solution Approach 2:
Noise fluorescence peaks from impurities are extracted and removed from the analysis. By identifying peaks that correspond to noise rather than actual base species, these noise peaks are taken out of the matrix conversion calculation, preventing them from causing inaccurate determination results while maintaining the accuracy of base species analysis.
2Measurement precision
If multiple wavelength bands are measured to identify fluorescent substance types, then the ability to distinguish between different fluorescent substances improves, but the complexity of data processing increases
Solution Approach 1:
Different wavelength bands are measured with different purposes: some bands are optimized for detecting specific fluorescent substance types while others are used for noise identification. The local quality of each wavelength band is utilized appropriately - certain bands provide high discrimination power for base species identification while others help characterize noise patterns, balancing the complexity of processing with improved distinction capability.
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 accurate identification and detection of biologically relevant elements like base species by removing noise fluorescence peaks, thereby improving the reliability of DNA sequencing results.
Implementation Method 1
The molecular weight separation is performed by electrophoresis using, for example, a polyacrylamide gel
Implementation Method 2
the main method is to fill a gel or a polymer that allows the molecular weight separation in a capillary and performs the electrophoresis
Implementation Method 3
the above-described fragments are preliminarily labeled with four types of fluorescent substances different by each terminal base species and irradiated with an excitation light at a specific detecting position, and the base species are determined from differences between the generated fluorescence spectra
Data Source
AI summary
There is a phenomenon where a noise in a spike shape caused by, for example, impurities and a noise peak having a spectrum different from a wavelength spectrum of a labeled fluorescent substance are detected during a capillary electrophoresis. Therefore, the disclosure provides a technique to identify an intensity of the labeled fluorescent substance itself without an effect by a noise fluorescence peak caused by impurities. In the disclosure, a fluorescence intensity property (a fluorescence profile of a noise) common to the noise peaks is set, the noise peak is handled as a fluorescent substance different from the labeled fluorescent substance, and the fluorescent substance and the noise are separated by color converting with the labeled fluorescent substance+the noise fluorescent substance (see FIG. 5).