Nanopore Throughput Speed Measurement Using Segmented Calibration
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Solution Overview
Problem
Current methods for characterizing the throughput properties of nanopores are inadequate, relying on macroscopic ionic current measurements that fail to provide a direct and unambiguous assessment of throughput speed, as they cannot distinguish between single and multiple objects passing through the pore and are convolved with temporal current behavior.
Innovation Solution
A method using a filiform calibration element with markers spaced at known distances, which interacts with the nanopore to produce detectable signals upon entry, exit, or within the pore, allowing for the determination of throughput speed by measuring time intervals or frequency of these interactions, and optionally complemented with ionic current monitoring.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If macroscopic ionic current measurements are used to characterize nanopore throughput, then the measurement can be performed with simple equipment, but the measurement precision is insufficient because it cannot distinguish between single and multiple objects passing through the pore
Solution Approach 1:
The calibration element is segmented into multiple markers spaced at known distances along its length. As each marker passes through the pore, it generates a discrete interaction event, allowing the system to measure throughput speed by timing the interval between successive events. This segmentation transforms a continuous measurement problem into discrete, distinguishable events that can be precisely timed and counted.
2Measurement precision
If ionic current monitoring is used to track objects through the pore, then continuous monitoring is possible, but the measurement becomes convolved with temporal current behavior making it difficult to extract accurate throughput speed
Solution Approach 1:
A calibration element with markers serves as an intermediary between the objects being studied and the detection system. The markers generate distinct interaction events (such as optical signals or mechanical blockades) that are easier to detect and time precisely than ionic current changes. This intermediary transforms the measurement into discrete events with clear temporal markers, eliminating the convolution problem inherent in continuous current monitoring.
3Productivity
If multiple objects pass through the pore simultaneously, then throughput increases, but the ionic current measurement cannot distinguish between single and multiple objects leading to measurement ambiguity
Solution Approach 1:
The calibration element contains multiple markers that act as copies or replicas along its length. As the calibration element passes through the pore, each marker generates a separate interaction event, creating a sequence of copies of the same signal pattern. By counting and timing these replicated events, the system can determine both the throughput speed and verify that objects are passing through individually, even at high throughput rates.
Data Source
AI summary
A method to determine the throughput speed v of a pore, comprising the steps of feeding, by means of a driving force F, a filiform calibration element through the pore, the calibration element having a plurality of markers spaced apart by known distances and configured to produce an interaction event that transmits a signal away from the pore upon interaction with the pore, detecting a plurality of interaction events, and determining a time interval Δt between successive interaction events, and/or a frequency ω of interaction events.