Amplicon-Based Temperature Calibration for Microfluidic Channels
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current microfluidic devices face challenges in precisely calibrating the temperature of multiple thermal control elements, which is essential for reliable PCR and thermal melt analysis, due to the complexity and cost of third-party calibration methods, and the need for accurate temperature uniformity across channels.
Innovation Solution
The method involves introducing amplicons with known melting temperatures into microfluidic channels, determining their melting temperatures, comparing them to expected values, and adjusting the thermal control elements accordingly, using synthetic or genomic DNA-based calibrants like Ultra Conserved Elements (UCEs) to ensure accurate calibration without interfering with sample amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If third-party calibration methods are used to calibrate thermal control elements, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The microfluidic device performs self-calibration using built-in thermal control elements and measurable thermal parameters. The system uses the known thermal characteristics of the microfluidic channel and reagents to automatically determine calibration parameters without requiring external third-party calibration services, thereby reducing complexity while maintaining precision.
Solution Approach 2:
The patent introduces thermal reagents or calibrant substances with known thermal properties as intermediaries to facilitate the calibration process. These intermediaries provide reference points for measuring thermal parameters, enabling accurate calibration through measurable thermal transitions without requiring complex external equipment.
2Measurement precision
If third-party calibration methods are used to calibrate thermal control elements, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The calibration data and thermal parameters are pre-determined through theoretical modeling and stored in the system. During actual operation, the system quickly retrieves and applies these pre-calculated calibration parameters, eliminating the need for time-consuming third-party calibration procedures while maintaining measurement precision.
Solution Approach 2:
The patent replaces physical third-party calibration procedures with computational methods. The system uses mathematical models and algorithms to calculate calibration parameters based on measured thermal data, substituting manual mechanical calibration processes with automated computational analysis that is both faster and equally precise.
3Manufacturing precision
If calibrants are used for temperature calibration, then manufacturing precision is improved, but they may interfere with sample amplification
Solution Approach 1:
The patent uses different calibrant substances for different thermal calibration points, each optimized for specific temperature ranges. The calibrants are selectively applied only where needed for calibration purposes in specific microfluidic channels or zones, rather than uniformly throughout the entire system, minimizing interference with sample amplification in other areas.
Solution Approach 2:
The calibration function is separated from the sample amplification function. The patent uses dedicated calibration channels or separate measurement protocols where calibrants are introduced only for calibration purposes, while sample amplification occurs in separate channels or during different time periods, eliminating cross-interference between the two functions.
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 approach allows for precise and efficient calibration of microfluidic channels, reducing downtime and maintaining high throughput by using calibrants that do not affect sample amplification, ensuring consistent and reliable temperature control for PCR and thermal melt analysis.
Implementation Method 1
amplicons with known melting temperatures into microfluidic channels, determining their melting temperatures
Data Source
AI summary
The present invention relates to the use of one or more amplicons as temperature calibrators. In some embodiments, the calibrators may be used to calibrate the temperature of a microfluidic channel in which amplification and/or melt analysis is performed. In some embodiments, the amplicons may be genomic, ultra conserved elements and/or synthetic. The amplicon(s) may have a known or expected melt temperature(s). The calibrators may be added to primers of study or may follow or lead the primers of study in the channel. The amplicon(s) may be amplified and melted, and the temperature(s) at which the amplicon(s) melted may be determined. The measured temperature(s) may be compared to the known temperature(s) at which the amplicon(s) was expected to melt. The difference(s) between the measured and expected temperatures may be used to calibrate/adjust one or more temperature control elements used to control and/or detect the temperature of the channel. In other embodiments, the UCE primers may function as a positive control to validate amplification has occurred.