Serial Flow Emulsion Processing to Reduce Cross-Contamination
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Solution Overview
Problem
Existing serial flow emulsion systems face issues with cross-contamination between individual volumes of the dispersed phase and/or the channel and/or tube containing the emulsion, particularly in processes like emulsion-based digital nucleic acid amplification, leading to inaccuracies and contamination.
Innovation Solution
The system employs an intake system, injector, partitioner, reactor, and detector configuration where the intake and process systems are separate, with an injector positioned between them, allowing for discrete aliquots of dispersed phase to be injected into the process system, and includes features like fluoropolymer surfaces, spacer fluids, and surfactants to minimize cross-contamination, along with optimized conduit connections and flow control to maintain laminar flow.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If serial flow emulsion processing is used to process multiple samples, then productivity is improved, but cross-contamination between samples increases
Solution Approach 1:
The system segments the continuous emulsion flow into discrete droplets using a partitioner, creating individually isolated reaction chambers. This segmentation prevents cross-contamination between samples while maintaining continuous processing capability, as each droplet acts as an independent reaction vessel that can be processed sequentially without contaminating other samples.
Solution Approach 2:
A continuous phase fluid serves as an intermediary carrier between the intake system and reaction chambers. This intermediary fluid allows for the transport of dispersed phase samples through the system while maintaining physical separation between different sample volumes, enabling serial processing without direct contact between samples.
2Device complexity
If the same conduit is used for multiple samples, then device complexity is reduced, but cross-contamination between samples increases
Solution Approach 1:
The conduit system is segmented into distinct zones: an intake conduit for receiving samples, a separation conduit with partitioner for creating discrete droplets, and reaction conduits for processing. This segmentation allows the same physical conduit to be used for multiple samples while preventing carryover through the droplet separation mechanism.
Solution Approach 2:
The system employs periodic injection of dispersed phase samples into the continuous flow, with spacing between injections. This periodic action, combined with the continuous phase carrier, ensures that samples are introduced in discrete, separated intervals, preventing overlap and contamination between consecutive samples.
3Productivity
If emulsion-based digital nucleic acid amplification is performed in serial flow, then productivity is improved, but measurement precision decreases due to cross-contamination
Solution Approach 1:
The partitioner creates physically separated droplets that serve as individual reaction chambers for nucleic acid amplification. This segmentation ensures that each droplet contains a discrete, known amount of sample material, enabling accurate digital quantitation while maintaining high processing throughput through serial flow.
Solution Approach 2:
The continuous phase acts as an intermediary that carries discrete droplets through the reaction system without allowing mixing between different sample volumes. This intermediary carrier maintains the integrity of each droplet's contents, ensuring measurement precision is maintained even at high processing speeds.
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 configuration significantly reduces cross-contamination to levels below 0.1%, ensuring accurate and reliable processing of multiple samples without sample-to-sample carryover or environmental interference, maintaining high precision in emulsion-based processes.
Implementation Method 1
surfactants to minimize cross-contamination
Implementation Method 2
fluoropolymer surfaces
Implementation Method 3
spacer fluids
Implementation Method 4
optimized conduit connections and flow control to maintain laminar flow
Data Source
AI summary
Disclosed herein are systems and methods for serial flow emulsion processes. Systems and methods as described herein result in reduced cross-contamination.