Multicapillary Electrophoresis Signal Normalization via Raman Reference
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Solution Overview
Problem
Multicapillary electrophoresis devices face challenges in making quantitative comparisons between capillaries due to variations in fluorescence intensity, despite methods like varying irradiation time and internal standard corrections, which fail to accurately normalize signals across multiple capillaries.
Innovation Solution
A multicapillary electrophoresis device with a capillary array, a light source, photodetector, and an arithmetic control unit that computes and corrects fluorescence signal intensities using a correction index determined for each capillary and fluorophore combination, and optionally measures Raman light to calculate this index.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a capillary array with multiple capillaries is used to accelerate analysis, then productivity is improved, but measurement precision deteriorates due to configurational variation in fluorescence intensity
Solution Approach 1:
The patent introduces Raman light as an intermediary reference signal to mediate between the excitation light source and the fluorescence detection. By measuring Raman light from the separation medium at the same wavelength as the excitation light, the system creates a reference that captures the actual excitation conditions for each capillary, enabling accurate normalization of fluorescence signals across multiple capillaries
Solution Approach 2:
The patent changes the parameter being measured from only fluorescence intensity to a combination of Raman light intensity and fluorescence intensity. By using the ratio of fluorescence intensity to Raman light intensity, the system compensates for variations in excitation light delivery to different capillaries, thereby normalizing the measurement parameter to enable accurate quantitative comparison
2Measurement precision
If internal standard reference material is used to correct fluorescence intensity, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent extracts the reference measurement function from the fluorescence detection system by using Raman light from the separation medium itself. Instead of adding complex internal standard materials to the samples, the system separates the reference measurement (Raman) from the analyte measurement (fluorescence), using the medium's own Raman signal as the reference
Solution Approach 2:
The separation medium serves itself as the reference standard by providing Raman light at the excitation wavelength. The same medium that performs the separation function also provides the reference signal for normalization, eliminating the need for separate internal standard materials and simplifying the overall system
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 quantitative comparisons between capillaries by normalizing fluorescence signal intensities, reducing variations and ensuring consistent analysis across multiple capillaries.
Implementation Method 1
Each DNA is fluorescently labeled and emits fluorescence by irradiation with excitation light
Implementation Method 2
a photodetector detecting fluorescence from a sample in each capillary
Implementation Method 3
an arithmetic control unit computing a signal intensity of the fluorescence according to a signal from the photodetector and corrects the signal intensity according to a correction index
Implementation Method 4
a correction index computation unit computing the correction index. The correction index computation unit irradiates the capillaries with excitation light and measures Raman light and computes the correction index based on a signal intensity of the Raman light
Data Source
AI summary
A multicapillary electrophoresis device includes: a capillary array formed by arranging a plurality of capillaries; a light source irradiating the capillaries with excitation light; a photodetector detecting fluorescence from a sample in the capillaries; and an arithmetic control unit calculating a signal intensity of the fluorescence according to a signal from the photodetector. The arithmetic control unit is configured so as to correct the signal intensity according to a correction index determined for each of the combinations of any of the capillaries and a fluorophore labeled to the sample.