Digital PCR Device Air Bubble Removal
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Solution Overview
Problem
In digital PCR, the presence of air bubbles affects the accuracy of fluorescence intensity and melting curve measurements, particularly when measuring mutant-type genes with small base sequence differences, leading to reduced measurement accuracy and gene type determination errors.
Innovation Solution
A digital PCR measuring apparatus that includes a temperature adjuster, a fluorescence measurement part, and a controller. The controller controls the temperature adjuster to increase the temperature, remove air bubbles, and then measure fluorescence intensity and melting curves with high accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If temperature is increased to measure melting curve, then DNA-probe hybridization stability is improved, but air bubbles are generated in the sample container
Solution Approach 1:
The patent applies preliminary action by performing a pre-measurement step before the actual melting curve measurement. The controller executes a preliminary measurement process that prepares the sample container by removing air bubbles through controlled temperature cycling and vibration, ensuring that the subsequent fluorescence intensity measurements are not interfered with by air bubbles. This preliminary preparation resolves the contradiction by eliminating the harmful effect before it can impact the main measurement process.
2Adaptability or versatility
If fluorescence measurement is performed at multiple wavelengths to determine multiple gene types, then gene type determination capability is improved, but measurement accuracy is reduced due to spectrum spread and light leakage
Solution Approach 1:
The patent applies the taking out principle by extracting and removing air bubbles from the measurement system before fluorescence measurement. By eliminating air bubbles that cause light scattering and interference, the system maintains high measurement precision even when performing multi-wavelength fluorescence measurements for multiple gene types. This allows the system to achieve both versatility in detecting multiple gene types and precision in fluorescence intensity measurement.
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 apparatus effectively removes the influence of air bubbles, enabling highly accurate measurement of fluorescence intensity and melting curves, which improves the accuracy of gene type determination.
Implementation Method 1
When air bubbles are generated in the vicinity of a sample as the temperature rises
Implementation Method 2
a fluorescently labeled probe that is hybridized to the target gene are used, and a melting curve of these is measured and analyzed
Data Source
AI summary
A digital PCR measuring apparatus capable of measuring a melting curve with high accuracy is provided. The digital PCR measuring apparatus includes a temperature adjuster that controls a temperature of a sample container including a plurality of minute regions, a fluorescence measurement part that measures fluorescence intensity of a plurality of minute regions, and a controller that controls the temperature adjuster and the fluorescence measurement part. The controller controls the temperature adjuster to raise a temperature of the sample container, and, after removing air bubbles generated in the sample container, measures fluorescence intensity of a plurality of minute regions while controlling the temperature adjuster to lower the temperature of the sample container, and measures a melting curve of a plurality of the minute regions.