Fluorescence Detection Instrument Multiplexing qPCR
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Solution Overview
Problem
Existing fluorescence detection instruments for multiplexing qPCR applications face challenges with bulky size, weight, and inadequate signal-to-noise ratio, requiring multiple optical devices and sequential sample placement, which complicates temperature and reaction time management.
Innovation Solution
A compact fluorescence detection instrument featuring a modular design with adjustable optical components, including a heating module, detecting module, and actuation module, allowing for simultaneous illumination and detection of multiple samples with improved signal-to-noise ratio through precise alignment and tilting of optical axes, and utilizing a combination of light sources and filters for efficient excitation and emission signal separation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple optical devices are employed for different excitation filters in multiplexing qPCR, then the detection capability for multiple fluorescent probes is improved, but the size, weight, and cost of the instrument increase significantly
Solution Approach 1:
A single optical device is designed to perform multiple functions by detecting different fluorescent signals through sequential excitation wavelengths. The system uses one light source that can emit multiple wavelengths and one detector that can detect multiple fluorescent emissions, replacing the need for multiple separate optical devices for different excitation filters.
Solution Approach 2:
Multiple optical components are merged into a single integrated optical device. The system combines the light source, excitation filter, and detector into one unit that can handle multiple excitation wavelengths and detect multiple fluorescent signals, thereby reducing the overall instrument size and weight.
2Adaptability or versatility
If sequential sample placement is used for detecting multiple fluorescent probes, then the detection of different excitation and fluorescence signals is improved, but the temperature and reaction time changes affect the detection results
Solution Approach 1:
The system maintains continuous detection of all samples simultaneously without sequential placement. All reaction containers remain in the detection area throughout the process, allowing continuous monitoring of fluorescent signals from multiple probes without removing or repositioning samples, thereby maintaining stable temperature and reaction conditions.
Solution Approach 2:
The optical system dynamically switches between different excitation wavelengths to excite different fluorescent probes while all samples remain in place. The single optical device can selectively excite different fluorophores at different wavelengths and detect their emissions, enabling multiplexing without physical sample movement.
3Device complexity
If a single optical device is used for multiplexing qPCR, then the instrument size and cost are reduced, but the signal-to-noise ratio must be sufficiently high to distinguish multiple fluorescent signals
Solution Approach 1:
Excitation filters are used as intermediaries to selectively transmit specific excitation wavelengths to the samples while blocking other wavelengths. This allows the single optical device to distinguish between different excitation sources and their corresponding fluorescent emissions, improving signal-to-noise ratio by preventing cross-excitation of different fluorophores.
Solution Approach 2:
The system exploits the different emission colors (wavelengths) of different fluorescent probes. By using a light source that can emit multiple wavelengths and a detector that can distinguish different emission wavelengths, the system can identify and measure multiple fluorescent signals simultaneously based on their characteristic color signatures, maintaining high signal-to-noise ratio.
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 instrument achieves a high signal-to-noise ratio of up to 347, reducing size and cost while enabling efficient multiplexing qPCR applications with flexible operation for single and multi-color detections, improving precision and convenience.
Implementation Method 1
After the light source at specific wavelength illuminates on the targeted nucleic acids, the DNA-binding fluorescent probes of the nucleic acids will react and enable fluorescent signals to be emitted
Implementation Method 2
Each excitation filter is able to transfer at least one excitation light at specific wavelengths from the light source to the assembly of multiple individual sites
Implementation Method 3
The second filter is located between the detecting module and the heating module
Data Source
AI summary
The disclosure relates to a fluorescence detection instrument, including a base, a heating module, a detecting module, an illumination module, and an actuation module. The heating module, the detecting module, and the actuation module are disposed on the base. The heating module includes plural heating holders, wherein each of the plural heating holders is adapted to accommodate a light-transmissive reaction container adapted to contain a fluorescent reaction mixture with at least one targeted fluorescent probe respectively. The detecting module is configured with the heating module to form plural detection channels, wherein the plural heating holders are located at the plural detection channels respectively. The actuation module is connected with the illumination module and adapted to drive the illumination module to move to at least one predetermined position to selectively match at least one combination of the heating holder on the corresponding detection channel.