Digital Nucleic Acid Dispersion Verification By Fluorescence Imaging
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing digital nucleic acid detection technologies face challenges in efficiently confirming even dispersion of nucleic acid samples in microwells or microdroplets, leading to detection errors and cumbersome experimental methods due to issues like bubbles, uneven volumes, and the need for additional indicators and detection systems.
Innovation Solution
A method involving the use of nucleic acid primers combined with fluorescent indicator molecules, excited by a specific excitation light source and detected using a fluorescence emission band, to determine even dispersion through fluorescence imaging, reducing the need for additional indicators and detection systems.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If additional non-nucleic acid amplification fluorescent indicator or specific nucleic acid with primer, probe, and fluorescent indicator is added to detect poor sample dispersion, then detection accuracy is improved, but device complexity and experimental burden increase
Solution Approach 1:
The fluorescent indicator molecules serve dual purposes: they act as both the nucleic acid amplification fluorescent indicator and the dispersion condition indicator. The same excitation light source and detector system are used for both nucleic acid detection and dispersion assessment, eliminating the need for separate detection systems and reducing device complexity while maintaining detection accuracy.
Solution Approach 2:
The patent combines the nucleic acid amplification detection function and the dispersion condition detection function into a single integrated system. By using the same fluorescent indicator, excitation light source, and detector for both purposes, the patent merges two separate detection processes into one, thereby reducing device complexity and experimental burden while ensuring accurate detection.
2Measurement precision
If scattering signal detection system is added to detect poor sample dispersion, then detection accuracy is improved, but device complexity and experimental burden increase
Solution Approach 1:
The fluorescent indicator molecules serve dual purposes: they act as both the nucleic acid amplification fluorescent indicator and the dispersion condition indicator. The same excitation light source and detector system are used for both nucleic acid detection and dispersion assessment, eliminating the need for separate detection systems and reducing device complexity while maintaining detection accuracy.
Solution Approach 2:
The patent combines the nucleic acid amplification detection function and the dispersion condition detection function into a single integrated system. By using the same fluorescent indicator, excitation light source, and detector for both purposes, the patent merges two separate detection processes into one, thereby reducing device complexity and experimental burden while ensuring accurate detection.
3Measurement precision
If additional detection systems are added to confirm sample dispersion, then measurement precision is improved, but ease of operation deteriorates
Solution Approach 1:
The fluorescent indicator molecules serve dual purposes: they act as both the nucleic acid amplification fluorescent indicator and the dispersion condition indicator. The same excitation light source and detector system are used for both nucleic acid detection and dispersion assessment, eliminating the need for separate detection systems and reducing device complexity while maintaining detection accuracy.
Solution Approach 2:
The patent combines the nucleic acid amplification detection function and the dispersion condition detection function into a single integrated system. By using the same fluorescent indicator, excitation light source, and detector for both purposes, the patent merges two separate detection processes into one, thereby reducing device complexity and experimental burden while ensuring accurate detection.
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 method effectively confirms even dispersion, reducing detection errors and experimental burden by allowing for efficient confirmation of nucleic acid sample distribution in microwells or microdroplets, thereby enhancing the convenience and efficiency of nucleic acid amplification reactions.
Implementation Method 1
fluorescence is excited using an excitation light source with a first fluorescence excitation band, and fluorescence images are detected using a detector for detecting a first fluorescence emission band for the nucleic acid reaction samples dispersed in the reaction areas
Data Source
AI summary
A confirming method for dispersion conditions of digital nucleic acid amplification reaction samples in reaction areas includes the following steps. The nucleic acid reaction samples are dispersed in the reaction areas. The nucleic acid reaction samples include nucleic acid primers and fluorescent indicator molecules for nucleic acid amplification. Parts of the nucleic acid primers are combined with the fluorescent indicator molecules for nucleic acid amplification before a digital nucleic acid amplification reaction. Next, fluorescence is excited using an excitation light source with a first fluorescence excitation band, and fluorescence images are detected using a detector for detecting a first fluorescence emission band for the nucleic acid reaction samples dispersed in the reaction areas. Afterwards, whether the nucleic acid reaction samples are evenly distributed in the reaction areas is determined using the fluorescence images.