Temperature-Dependent Fluorophores for Nucleic Acid Sequencing
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Solution Overview
Problem
Current nucleic acid sequencing technologies relying on fluorescence signal detection require complex and expensive optics, limiting throughput and efficiency in DNA sequencing.
Innovation Solution
The use of thermally-dependent fluorophores that emit light at different intensities over specific temperature ranges, allowing for the determination of incorporated nucleotides with fewer chemistry steps and simpler optics by adjusting the temperature during sequencing cycles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If conventional fluorescence-based sequencing is used, then base detection can be achieved, but complex and expensive optics are required
Solution Approach 1:
The patent changes the detection parameter from wavelength (color) to temperature. By using fluorophores whose emission intensity is temperature-dependent rather than wavelength-dependent, the system can distinguish between different nucleotides through temperature variations instead of complex wavelength filtering optics. This parameter change fundamentally simplifies the optical detection system while maintaining base detection capability.
Solution Approach 2:
The patent extracts and eliminates the complex wavelength filtering optics from the sequencing system. By replacing the conventional approach that requires multiple optical filters and dichroic mirrors with a temperature-based detection method, the system removes unnecessary optical components while preserving the ability to detect incorporated nucleotides.
2Productivity
If conventional fluorescence detection with multiple optical filters is used, then nucleotide distinction is achieved, but throughput is limited
Solution Approach 1:
The patent changes the detection parameter from wavelength to temperature, enabling faster detection cycles. By using temperature-dependent fluorescence intensity changes rather than wavelength filtering, the system can rapidly distinguish between nucleotides without the time-consuming optical filter switching required in conventional systems, thereby increasing throughput.
Solution Approach 2:
The patent replaces the mechanical optical filtering system with a thermal detection method. Instead of using physical optical filters that must be switched or tuned to detect different wavelengths, the system uses temperature variations to modulate fluorescence intensity, substituting a thermal field for a mechanical-optical field and enabling faster detection.
3Illumination intensity
If high-power lasers and large-area flow cells are used, then sufficient fluorescence signals are generated, but cost and complexity increase
Solution Approach 1:
The patent changes the signal detection parameter from wavelength-specific intensity (requiring high-power lasers) to temperature-dependent intensity ratios. By measuring changes in fluorescence intensity relative to temperature rather than absolute intensity at specific wavelengths, the system can achieve sufficient signal discrimination without requiring high-power laser illumination, thereby reducing optical system complexity and cost.
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 enables efficient DNA sequencing with reduced need for complex optics, achieving higher throughput and accuracy by utilizing temperature-dependent fluorescence to distinguish between nucleotides without the need for wavelength filtering.
Implementation Method 1
The selected fluorophores emit light in a temperature-dependent manner over a range of temperatures between room temperature and an upper temperature (e.g., 100° C.).
Data Source
AI summary
Disclosed herein are improved methods and systems for sequencing nucleic acid that exploit the temperature-dependence of the emitted intensity of fluorescent dyes. The temperature of the sequencing reaction is adjusted during each sequencing cycle, and the emission, or lack of emission, of light meeting or exceeding a threshold by the fluorescent dyes at different temperatures, or within different temperature ranges, is used to detect the fluorescent labels of the incorporated dNTPs and thereby sequence the nucleic acid. The disclosed methods enable a determination of the dNTP incorporated at any given site with a reasonable number of chemistry steps without the complex optics necessary for prior-art systems.