Multiplex PCR Fluorescence Detection Without Filter Wheels
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Solution Overview
Problem
Conventional real-time PCR instruments face challenges in efficiently detecting multiple target species using fluorescent dyes due to complex electromechanical parts and limited spectral resolution, leading to reduced accuracy and increased operating time.
Innovation Solution
A multiplex fluorescence detection device with a plurality of optical modules optimized for discrete wavelength bands, coupled via a multi-legged optical fiber bundle to a single detector, and a laser valve control system for precise fluid flow management, allowing for simultaneous detection of multiple reactions and independent temperature modulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional real-time PCR instruments use white light sources with filter wheels for spectrally resolving fluorescent dyes, then detection capability is achieved, but device complexity increases and reliability decreases due to complicated electromechanical parts
Solution Approach 1:
The patent replaces the mechanical filter wheel system with an optical fiber bundle that delivers light from a single source to multiple detection channels. This eliminates the moving mechanical parts (filter wheel, motors, bearings) while maintaining the ability to resolve multiple fluorescent dye signals through optical path separation rather than mechanical filtering.
Solution Approach 2:
The single light source serves multiple functions by illuminating multiple reaction chambers simultaneously through the optical fiber bundle. The system achieves multi-wavelength detection capability without requiring multiple separate light sources or mechanical filtering components, as each optical channel can be independently optimized for specific fluorescent dyes.
2Adaptability or versatility
If conventional real-time PCR instruments use filter wheels for spectral resolution, then multiple fluorescent dyes can be detected, but operating time increases due to mechanical movement and positioning
Solution Approach 1:
The patent eliminates the mechanical filter wheel positioning system and replaces it with a static optical fiber bundle configuration. Spectral resolution is achieved through optical path separation and wavelength-specific detection in parallel channels, eliminating the time required for mechanical movement and positioning of filters during operation.
Solution Approach 2:
The system enables continuous simultaneous detection of multiple fluorescent dyes across all reaction chambers without interruption. The static optical configuration allows all detection channels to operate continuously in parallel, eliminating the sequential filtering process that would require mechanical movement and time delays between measurements.
3Measurement precision
If conventional PCR instruments use complicated electromechanical parts for dye detection, then spectral separation is achieved, but reliability decreases due to susceptibility to wear
Solution Approach 1:
The patent replaces the electromechanical filter wheel assembly with a purely optical static system using fiber optics. This eliminates components subject to mechanical wear (motors, bearings, gears, belts) while maintaining spectral separation capability through optical path design. The system has no moving parts in the detection pathway, significantly improving reliability and reducing maintenance requirements.
4Measurement precision
If conventional real-time PCR instruments detect multiple target species, then detection accuracy is improved, but device complexity and operating time increase
Solution Approach 1:
The patent merges multiple detection functions into a single integrated optical system. The optical fiber bundle combines light delivery to multiple chambers and simultaneous collection from multiple channels into one unified structure, eliminating the need for separate mechanical filtering systems for each wavelength. This consolidation maintains multi-species detection accuracy while reducing overall system complexity.
Solution Approach 2:
The single optical fiber bundle system performs multiple functions simultaneously: delivering excitation light to multiple reaction chambers, collecting fluorescent signals from multiple dyes, and enabling parallel detection across all channels. This multi-functional design achieves accurate multiplex detection without requiring separate specialized components for each detection task.
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
The solution enables accurate, real-time detection of multiple target species with reduced operating time and increased accuracy, supporting various biological reactions and allowing for portable operation in remote settings.
Implementation Method 1
an energy source that outputs electromagnetic energy at a first level to determine a position of the disk and at a second level to open the valve
Implementation Method 2
emits a laser beam that is detected by sensor through a slot in the disk to map the disk position with respect to the rotating platform
Implementation Method 3
the laser control valve system may focus higher-energy laser light on a valve to open the valve and allow contents to flow
Implementation Method 4
The plurality of optical modules may be optically coupled to a single detector by a multi-legged optical fiber bundle
Implementation Method 5
Each of the optical modules may be optimized for detection of a respective fluorescent dye at a discrete wavelength band
Data Source
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AI summary
Techniques are described for the detection of multiple target species in real-time PCR (polymerase chain reaction). For example, a system comprises a data acquisition device and a detection device coupled to the data acquisition device. The detection device includes a rotating disk having a plurality of process chambers having a plurality of species that emit fluorescent light at different wavelengths. The device further includes a plurality of removable optical modules that are optically configured to excite the species and capture fluorescent light emitted by the species at different wavelengths. A fiber optic bundle coupled to the plurality of removable optical modules conveys the fluorescent light from the optical modules to a single detector. In addition, the device may control the flow of fluid in the disk by locating and selectively opening valves separating chambers by heating the valves with a laser.