Single-Channel Fluorescence Sequencing via Amplitude-Modulated Base Calling
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Solution Overview
Problem
Existing DNA sequencing systems require multiple excitation light sources and detectors, which increase sequencing time and expose samples to unnecessary light, leading to light-induced DNA damage and reduced sequencing speed.
Innovation Solution
A nucleic acid sequencing system utilizing a single light source and a single detector, employing amplitude modulation to encode nucleobases with different fluorescence intensities, allowing identification of four types of nucleobases in a single optical channel.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple excitation light sources and detectors are used, then measurement precision is improved, but device complexity increases and sequencing speed decreases
Solution Approach 1:
The patent merges multiple optical channels into a single detection channel by using fluorophores with different quantum yields that all emit at the same wavelength. This allows four types of nucleobases to be distinguished through amplitude modulation of fluorescence intensity in a single channel, eliminating the need for multiple detectors and light sources while maintaining identification accuracy
Solution Approach 2:
The patent changes the parameter being measured from wavelength (spectral differentiation) to intensity (amplitude modulation). By using fluorophores with different quantum yields, the system encodes nucleobase identity in fluorescence intensity levels rather than emission wavelengths, enabling single-channel detection of four nucleobase types
2Measurement precision
If multiple excitation light sources are used, then measurement precision is improved, but loss of time increases due to longer exposure requirements
Solution Approach 1:
The patent enables continuous fluorescence detection at a single wavelength by using fluorophores with different quantum yields. This allows for faster imaging because the system does not need to sequentially scan multiple wavelengths or channels, reducing imaging time while maintaining the ability to distinguish four nucleobase types through intensity variations
3Productivity
If exposure time is reduced to increase sequencing speed, then productivity is improved, but signal-to-noise ratio deteriorates
Solution Approach 1:
The patent changes from spectral resolution to intensity resolution, using fluorophores with different quantum yields that all emit at the same wavelength. This amplitude modulation approach allows for faster exposure times because the signal differentiation is based on intensity levels rather than wavelength separation, improving sequencing speed while maintaining signal-to-noise ratio through the use of a single optimized detection channel
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 reduces imaging time, simplifies optics, lowers instrument cost, and improves signal-to-noise ratio, enabling faster sequencing with reduced light exposure and lower computational requirements.
Implementation Method 1
excite deoxyribonucleic acid analogs conjugated with fluorescent labels in a target polynucleotide
Data Source
AI summary
The disclosed technology relates to systems and methods for nucleic acid sequencing utilizing a single light source and a single detector. The disclosed technology may use the light source to stimulate a fluorescence emission from a polynucleotide and identify a nucleobase in the polynucleotide based on the intensity of the fluorescence emission received by the detector. The disclosed technology may utilize four types of nucleotide analogs which emit light at four distinguishable levels when excited by the light source. In various embodiments, the four types of nucleotide analogs may be coupled to different fluorophores or the same fluorophore with different probabilities or copy numbers.