PCR Fluorescence Detection Using a Spectrograph for Multi-Marker Assays
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Solution Overview
Problem
Existing PCR detection methods are limited by the need for optical filters to detect multiple wavelengths, which restricts simultaneous detection of multiple fluorescence markers and can lead to uneven distribution of fluorescent substances, affecting detection efficiency and accuracy.
Innovation Solution
A PCR detector utilizing a spectrograph to detect emitting light with different wavelengths simultaneously, eliminating the need for optical filters and incorporating a light source rotating device to ensure uniform distribution of light spots, combined with a method for calculating the Ct value through fluorescence intensity normalization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a conventional PCR detector is used, then the device structure is relatively simple, but the detection precision and reliability are insufficient due to temperature non-uniformity in the well block
Solution Approach 1:
The heating system is segmented into multiple independent heating units (first heating unit, second heating unit, third heating unit, fourth heating unit) positioned at different locations of the well block. Each heating unit can be independently controlled to provide localized heating, enabling precise temperature control across different regions of the well block and eliminating temperature non-uniformity issues.
Solution Approach 2:
Different heating units are applied to different locations of the well block based on specific thermal requirements. The controller adjusts each heating unit's output independently to achieve uniform temperature distribution across the well block, ensuring that each region maintains the optimal temperature for its specific PCR reaction conditions.
2Stability of the object's composition
If temperature control is improved with multiple heating units, then temperature uniformity improves, but the control system becomes more complex
Solution Approach 1:
The controller serves multiple functions: it controls all heating units, monitors temperature through the sensor, processes control signals, and coordinates the operation of different heating zones. This multi-functional design consolidates control complexity into a single integrated unit rather than requiring separate control systems for each heating element.
Solution Approach 2:
The temperature sensor provides real-time feedback on the well block temperature to the controller. The controller uses this feedback information to dynamically adjust the heating units' output, creating a closed-loop control system that automatically maintains temperature uniformity without requiring complex manual intervention or oversight.
3Speed
If heating efficiency is increased, then the heating speed improves, but energy consumption increases
Solution Approach 1:
The heating units operate in periodic cycles rather than continuously. The controller activates heating units only when temperature increase is needed based on feedback from the temperature sensor, and deactivates them when the target temperature is reached or maintained. This periodic operation reduces overall energy consumption while maintaining effective heating speed when required.
Solution Approach 2:
The heating system dynamically adjusts the power output of individual heating units based on real-time temperature conditions. The controller modulates heating intensity to match the actual thermal needs of the well block, preventing energy waste from excessive heating while ensuring rapid temperature changes when required by the PCR protocol.
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 allows for simultaneous detection of multiple fluorescence markers, enhances detection efficiency, improves accuracy, and simplifies the detector's structure by eliminating optical filters, while ensuring reliable Ct value calculation.
Implementation Method 1
a heating unit is provided in the well block, wherein the heating unit is configured to heat the well block to a predetermined temperature
Implementation Method 2
a temperature sensor is provided in the well block, wherein the temperature sensor is configured to detect temperature of the well block
Data Source
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AI summary
A PCR detector is provided, and belongs to the technical field of PCR detection. The PCR detector includes an excitation light source module, a chip device and a detection part; a reaction bin is arranged in an emitting direction of excitation light of the excitation light source module, the detection part is arranged at one side of the reaction bin, and the emitting light formed after the excitation light illuminates the reaction bin can be detected by the detection part, wherein an emitting direction of the excitation light is located below the detection part, and the detection part is configured to detect emitting light emitted by the detected sample in a vertical direction due to the illumination of the excitation light; and the detection part includes a spectrograph, a wavelength scope of a spectrum detected by the spectrograph is 340-850nm, and the spectrograph can detect the excitation light and the emitting light. The spectrograph may detect different multi-channel spectrum at the same time, thereby realizing detecting a plurality of fluorescence marks at the same time, namely, detecting various different nucleic acids at the same time.