Multi-Capillary Fluorescence Binning for Sensitivity and Dynamic Range
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Solution Overview
Problem
Existing capillary electrophoresis instruments struggle to achieve both high sensitivity and high dynamic range when analyzing multiple types of fluorophores, as methods combining high-sensitivity and low-sensitivity modes fail to accurately identify and quantify multiple fluorophores.
Innovation Solution
A multi-capillary electrophoresis instrument that controls the composition of the electrophoresis separation medium, laser beam settings, 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
1Duration of action of moving object
If methods combining high-sensitivity and low-sensitivity modes are used to expand dynamic range, then the dynamic range is improved, but the ability to accurately identify and quantify multiple fluorophores deteriorates
Solution Approach 1:
The patent applies dynamics by making the binning configuration adjustable and adaptable. The system dynamically optimizes the binning parameters (number of bins, pixels per bin) based on the specific measurement requirements, allowing transition between different sensitivity configurations without physical hardware changes. This enables the system to maintain high measurement precision while achieving expanded dynamic range through software-controlled parameter adaptation.
Solution Approach 2:
The patent utilizes parameter changes by optimizing the binning parameters (number of bins, pixels per bin, wavelength ranges) to achieve both high sensitivity and high dynamic range simultaneously. By carefully selecting and adjusting these parameters, the system can resolve multiple fluorophores accurately while handling a wide concentration range, eliminating the need to switch between high-sensitivity and low-sensitivity modes.
2Measurement precision
If the number of pixels per bin is increased to improve sensitivity, then the sensitivity is improved, but the dynamic range deteriorates
Solution Approach 1:
The patent applies segmentation by dividing the detector pixels into multiple bins, where each bin can be independently configured with optimal pixel counts. This segmentation allows different wavelength regions to have different bin configurations, enabling high sensitivity in regions where fluorophores are present while maintaining dynamic range through appropriate pixel allocation. The segmented bin structure resolves the contradiction by allowing localized optimization rather than uniform configuration.
Solution Approach 2:
The patent implements local quality by assigning different numbers of pixels to different bins based on the specific measurement requirements for each wavelength region. Instead of using a uniform pixel allocation, the system optimizes the pixel count for each bin locally, allocating more pixels to bins detecting weak signals (high sensitivity) and fewer pixels to bins detecting strong signals (maintaining dynamic range). This local optimization resolves the sensitivity-dynamic range contradiction.
3Productivity
If multiple fluorophores are analyzed simultaneously, then the productivity is improved, but the measurement precision deteriorates due to signal overlap
Solution Approach 1:
The patent applies dimensionality change by utilizing the wavelength dimension to resolve fluorophore overlap. By dispersing light across multiple wavelength bins and analyzing the spectral distribution, the system can distinguish between multiple fluorophores even when their emission spectra overlap. This spectral dimension allows simultaneous analysis of multiple fluorophores with high precision, maintaining both productivity and measurement accuracy.
Solution Approach 2:
The patent utilizes feedback through iterative optimization of binning parameters based on the detected signal characteristics. The system analyzes the spectral data and adjusts the bin configuration to optimize the separation and quantification of multiple fluorophores. This feedback mechanism ensures that even when fluorophores are analyzed simultaneously, the measurement precision is maintained by adapting the binning parameters to the actual spectral distribution.
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 with a wide concentration range without concentration adjustment, allowing for the identification and quantification of multiple fluorophores in a single mode.
Implementation Method 1
a laser beam is incident perpendicularly to each axis of E (E is an integer of 1 or more) capillaries arranged on the same plane and irradiates them simultaneously. As a result, E light-emitting points arranged on a straight line are formed on the E capillaries arranged on the same plane. When DNA fragments labeled with G (G is an integer of 1 or more) 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 by a second camera lens.
Implementation Method 3
On the image sensor, the E wavelength-dispersed images of the fluorescences from the E light-emitting points are arranged in parallel to each other without being mixed with each other. Therefore, the fluorescence from each light-emitting point can be spectroscopically measured independently.
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.