Fluorescence Detection Phase Control for PCR Noise Reduction
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Solution Overview
Problem
In PCR reaction processing apparatuses with multiple fluorescence detection devices, interference occurs due to overlapping wavelength ranges of excitation and fluorescence light, leading to noise and reduced sensitivity in fluorescence measurements.
Innovation Solution
A reaction processing apparatus with a channel for sample movement, featuring multiple fluorescence detection devices where the excitation lights from different devices flash at a predetermined duty ratio with a phase difference within specific ranges to minimize interference, and optical heads with numerical apertures between 0.07 to 0.23, ensuring proper spacing and alignment to reduce noise.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple fluorescence detection devices are arranged in the channel extending direction to detect fluorescence from samples passing through a single channel, then the detection capability for multiple fluorescent dyes is improved, but interference between detection devices occurs due to overlapping wavelength ranges
Solution Approach 1:
The patent applies periodic action by making excitation lights from multiple fluorescence detection devices flash alternately at predetermined duty ratios with specific phase differences. This temporal separation allows each detection device to operate in distinct time windows, preventing wavelength overlap interference while maintaining the ability to detect multiple fluorescent dyes simultaneously through the same channel.
2Object-affected harmful factors
If excitation lights from multiple fluorescence detection devices are made to flash with predetermined duty ratios and phase differences, then interference between devices is minimized, but the system complexity increases
Solution Approach 1:
The patent employs dynamics by implementing controllable flashing mechanisms with adjustable duty ratios and phase differences for excitation lights. This dynamic control allows the system to optimize interference reduction while maintaining operational flexibility. The temporal modulation of excitation lights creates a manageable complexity structure that can be controlled through software or simple circuitry.
3Measurement precision
If optical heads with specific numerical apertures (0.07 to 0.23) are used with proper spacing and alignment, then noise is reduced and measurement sensitivity is improved, but the device configuration becomes more complex
Solution Approach 1:
The patent applies parameter changes by specifying optimal numerical aperture ranges (0.07 to 0.23) for optical heads and defining precise spacing and alignment parameters. These parameter optimizations reduce noise and improve measurement sensitivity. By establishing specific parameter ranges rather than requiring complex adaptive systems, the patent achieves high measurement precision with manageable configuration complexity.
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 configuration enhances the sensitivity of fluorescence measurements by minimizing noise and interference, allowing for accurate real-time monitoring of DNA amplification during PCR.
Implementation Method 1
a first fluorescence detection device that irradiates a sample inside a first fluorescence detection region set in the channel with first excitation light and also detects first fluorescence produced from the sample by the irradiation with the first excitation light; and a second fluorescence detection device that irradiates a sample inside a second fluorescence detection region set in the channel with second excitation light and also detects second fluorescence produced from the sample by the irradiation with the second excitation light
Data Source
AI summary
A reaction processing apparatus includes: a reaction processing vessel; a first fluorescence detection device that irradiates a sample with first excitation light and detects first fluorescence produced from the sample; and a second fluorescence detection device that irradiates a sample with second excitation light and detects second fluorescence produced from the sample. The wavelength range of the first fluorescence and the wavelength range of the second excitation light overlap at least partially. The first excitation light and the second excitation light flash at a predetermined duty ratio d. The phase difference between the flashing of the first excitation light and the flashing of the second excitation light is set within a range of 2π(pm−Δpm) (rad) to 2π(pm+Δpm) (rad) or within a range of 2π[(1−pm)−Δpm] (rad) to 2π[(1−pm)+Δpm] (rad), where pm=d−d2 and Δpm =0.01*pm.