Nanopore Arrays With Opaque Layers for Background Noise Reduction
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nanopore sequencing technologies face challenges in reducing background noise during optical detection of single molecules, particularly due to light transmission through nanopore arrays, which complicates and expenses microscopy systems.
Innovation Solution
The implementation of nanopore arrays with opaque layers that block or absorb excitation light, reducing background noise by preventing light from passing through the signal generation regions, allowing for direct illumination of optical labels and minimizing nonspecific fluorescence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If direct illumination systems are used to excite fluorescent labels, then detection sensitivity is improved, but background noise increases due to light transmission through the nanopore array
Solution Approach 1:
The system segments the illumination path by introducing an opaque layer that divides the nanopore array into illuminated and non-illuminated regions. Only the signal generation region adjacent to the opaque layer receives excitation light, while other regions remain dark. This spatial segmentation allows direct illumination to excite fluorescent labels with high intensity while preventing light from transmitting through the entire array and creating background noise.
Solution Approach 2:
The opaque layer creates local quality differences across the nanopore array by providing selective optical absorption. The region adjacent to the opaque layer has high light absorption properties, creating a localized excitation zone with intense illumination for sensitive detection. Other regions maintain low light transmission, minimizing background noise. This local quality approach enables high detection sensitivity without pervasive background interference.
2Object-affected harmful factors
If opaque layers are added to block light transmission, then background noise is reduced, but device complexity increases
Solution Approach 1:
The harmful light transmission is extracted and removed from the system by introducing an opaque layer that selectively blocks excitation light. This simple additive component extracts the problematic light transmission without requiring complex microscopy systems or multiple optical elements. The opaque layer serves as a straightforward light-blocking element that reduces background noise while adding minimal structural complexity.
Solution Approach 2:
The opaque layer functions as a simple, inexpensive light-blocking component that can be integrated into the nanopore array substrate. Rather than requiring expensive, complex microscopy systems with multiple optical elements and alignment mechanisms, the solution uses a basic opaque material layer that provides effective light blocking. This approach trades minimal, simple structural addition for significant reduction in background noise, avoiding the need for complex imaging infrastructure.
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 effectively reduces background noise in optical signals, enabling more accurate and cost-effective single molecule analysis by preventing light from exciting materials outside the intended reaction sites, thus enhancing the reliability of sequence determination in nanopore sequencing.
Implementation Method 1
an opaque layer co-extensive therewith, the nanopore array comprising a plurality of apertures and separating a first chamber and a second chamber... wherein the opaque layer substantially prevents light from passing through the nanopore array
Implementation Method 2
different kinds of nucleotides of the polynucleotides are labeled with different fluorescent labels that generate distinguishable fluorescent signals... exciting with an excitation beam the fluorescent labels of the polynucleotides as they translocate through the signal generation regions
Data Source
Figure 1A
Figure 1B
Figure 1C
AI summary
The invention is directed to nanopore arrays comprising opaque layers that reduce background fluorescence in optical signal collected in applications of such arrays for analyzing molecules. In some embodiments, such arrays are used to determine characteristics of polymers, such as polynucleotides, in methods comprising the steps of translocating polymers through nanopores of such arrays wherein polymers have one or more optical labels, exciting optical labels of the polymers in a signal generation region of each nanopore extending from the opaque layer toward the direction of the excitation beam, detecting optical signals from the signal generation regions of each nanopore to determine characteristics of the polymer translocating therethrough.