Multi-Capillary Electrophoresis Binning for Fluorophore Dynamic Range
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Solution Overview
Problem
Existing methods for achieving high sensitivity and high dynamic range in capillary electrophoresis instruments fail when analyzing multiple types of fluorophores, as they cannot accurately identify and quantify the concentrations of each fluorophore due to spectral crosstalk and saturation issues.
Innovation Solution
A multi-capillary electrophoresis instrument that controls the composition of the electrophoresis separation medium, laser beam, optical system, exposure time, and bin settings, along with optimal pixel configurations on the image sensor, to achieve both high sensitivity and high dynamic range by setting the number of pixels and software binning within predetermined ranges.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If wavelength dispersion is used to measure multiple fluorophores, then measurement precision is improved, but device complexity increases due to the need for precise optical system configuration and bin settings
Solution Approach 1:
The patent optimizes specific parameters including the number of pixels assigned to each wavelength bin, exposure time, and bin settings to achieve the optimal balance between sensitivity and dynamic range. By adjusting these parameters within predetermined ranges, the system maintains high measurement precision while managing device complexity.
2Measurement precision
If exposure time is increased to improve sensitivity, then measurement precision is improved, but dynamic range is reduced due to signal saturation
Solution Approach 1:
The patent implements dynamic adjustment of exposure time based on the concentration range being measured. The system selects appropriate exposure times from predetermined settings to optimize both sensitivity and dynamic range, avoiding fixed exposure time limitations. This dynamic approach allows the system to adapt to different concentration levels and maintain optimal performance across the entire dynamic range.
3Productivity
If multiple fluorophores are analyzed simultaneously, then productivity is improved, but measurement precision deteriorates due to spectral crosstalk between fluorophores
Solution Approach 1:
The patent divides the wavelength spectrum into distinct bins, with each bin assigned to a specific fluorophore. This segmentation prevents spectral crosstalk by ensuring that each fluorophore's signal is measured in its dedicated wavelength range. The system simultaneously measures multiple fluorophores in parallel across different bins, maintaining both high productivity and measurement precision.
4Measurement precision
If bin width is decreased to improve wavelength resolution, then measurement precision is improved, but signal intensity is reduced due to fewer pixels per bin
Solution Approach 1:
The patent introduces a second dimension of optimization by adjusting the number of pixels assigned to each bin while maintaining appropriate bin width. Instead of relying solely on bin width for resolution, the system uses a combination of bin width and pixel allocation to achieve both high wavelength resolution and sufficient signal intensity. This multi-dimensional approach allows simultaneous optimization of resolution and signal strength.
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 analysis of multiple fluorophores across a wide concentration range without concentration adjustment, allowing for the identification and quantification of each fluorophore's concentration effectively.
Implementation Method 1
When DNA fragments labeled with G types of fluorophores passes through each light-emitting point on each capillary by electrophoresis, the fluorophores are excited by the laser beam irradiation and emit fluorescences
Implementation Method 2
E collimated fluorescences are dispersed by wavelength through a transmission-type diffraction grating, and E wavelength-dispersed images are formed on an image sensor
Implementation Method 3
a capillary electrophoresis instrument for DNA analysis, a laser beam is incident perpendicularly to each axis of E capillaries arranged on the same plane and irradiates them simultaneously
Data Source
AI summary
In an instrument configured to spectroscopically divide fluorescences emitted from a plurality of capillaries and collectively measure the fluorescences using an image sensor, when the number of pixels of a binning region on the image sensor on which a predetermined wavelength-band component of each fluorescence is projected is denoted by Bm, the number of pixels of hardware binning is denoted by Bh, the number of pixels of software binning is denoted by Bs, Bm=Bh×Bs, the total noise measured in a case where Bm=Bh=Bs=1 is denoted by N, the readout noise is denoted by Nr, the dark-current noise is denoted by Nd, and the shot noise is denoted by Ns, Bm, Bh, Bs, N, Nr, Nd, and Ns satisfy a predetermined relationship, thereby realizing high sensitivity and high dynamic range in fluorescence measurement.