Microfluidic Droplet Generation for Uniform Sample Discretization

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Solution Overview

Problem

Current methods for discretizing sample volumes are limited by their complexity, cost, and inefficiency, particularly in generating small, uniform volumes suitable for biochemical assays, as they often require specialized equipment and are not well-suited for continuous monitoring or handling very small volumes.

Innovation Solution

A fluidic device that partitions samples based on interplay between fluidic forces, interfacial tension, and channel geometry, allowing for simple, robust, and versatile discretization of sample volumes into localized arrays, which can be easily manipulated and analyzed, using minimal fluid interconnects and straightforward flow geometry.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If traditional methods like nebulizers or emulsion mixing are used to discretize sample volumes, then small individual volumes can be generated, but the droplets exhibit large size variations and are difficult to individually manipulate and analyze

Engineering Contradiction:
Improvedroplet size uniformityVSAvoidindividual droplet manipulation
Core Design Contradiction:
Manufacturing precisionVSEase of operation

Solution Approach 1:

The continuous sample flow is segmented into discrete droplets by an immiscible carrier fluid flowing through a microfluidic channel. The channel geometry and flow rates are controlled to produce monodisperse droplets of uniform size, which can then be individually manipulated and analyzed in subsequent steps

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

An immiscible carrier fluid acts as an intermediary substance that encapsulates the sample into discrete droplets. This carrier fluid enables the sample to be discretized into uniform volumes while maintaining stability and facilitating individual manipulation through subsequent electrowetting or dielectrophoretic forces

Inventive Principle:
Principle #24Intermediary (Mediator)

2Manufacturing precision

If microwell platforms with manual or robotic dispensing are used, then spatially localized samples can be created, but the process is tedious, expensive, and difficult for volumes below 0.5μL

Engineering Contradiction:
Improvespatial localization accuracyVSAvoiddispensing equipment requirements
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

Traditional mechanical dispensing systems are replaced with a microfluidic system that uses controlled fluid flow and interfacial tension to generate and position droplets. Electrowetting or dielectrophoretic forces are then used to manipulate the droplets into desired spatial arrangements, eliminating the need for complex robotic pipettes and enabling handling of sub-0.5μL volumes

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The system changes key parameters including flow rates of sample and carrier fluid, channel geometry dimensions, and applied electric field strengths to control droplet formation, size, and positioning. By adjusting these parameters, the system achieves precise spatial localization without requiring complex dispensing equipment

Inventive Principle:
Principle #35Parameter changes

3Ease of operation

If electrowetting is used for droplet manipulation, then droplet positioning is improved, but it mostly works for samples above the nanoliter scale and requires sophisticated electrode patterns

Engineering Contradiction:
Improvedroplet positioning controlVSAvoidelectrode pattern complexity
Core Design Contradiction:
Ease of operationVSDevice complexity

Solution Approach 1:

Instead of using complex sophisticated electrode patterns, the invention employs simpler electrode arrangements that apply electric fields locally to specific droplets. The electrodes are positioned to create targeted electric field gradients that enable individual droplet manipulation through electrowetting or dielectrophoresis, reducing overall device complexity while maintaining positioning control

Inventive Principle:
Principle #3Local quality

4Manufacturing precision

If patterned SAMs are used to generate smaller droplet volumes, then uniform nanoliter-scale droplets can be formed, but surface preparation is labor intensive and the substrate is often opaque

Engineering Contradiction:
Improvedroplet volume uniformityVSAvoidsurface preparation complexity
Core Design Contradiction:
Manufacturing precisionVSEase of manufacture

Solution Approach 1:

The invention extracts the droplet formation process from the substrate surface itself, using a microfluidic channel system where droplets are formed by fluid flow control rather than by surface patterning. This eliminates the need for labor-intensive SAM preparation and gold coating, and allows use of transparent substrates for optical monitoring while still achieving uniform nanoliter-scale droplet volumes

Inventive Principle:
Principle #2Taking out (Extraction)

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

The device enables efficient and economical generation of uniform sample volumes, suitable for various applications including PCR, genotyping, and biomedical assays, with the ability to combine different manipulation and detection methods, reducing sample loss and increasing analytical flexibility.

Implementation Method 1

A fluidic device partitions samples based on interplay between fluidic forces, interfacial tension, and channel geometry

Methodology Applied
Scientific EffectInterfacial tension: Surface Tension

Implementation Method 2

The discretized volumes reside within the sample compartments. The spacing in a large array of samples is predefined by the spacing of the device sample compartments

Methodology Applied
Scientific EffectCapillary action: Capillary Action

Data Source

PatentEP2324352B1Method and apparatus for the discretization and manipulation of sample volumes
Publication Date: 2020.06.03 UNIV OF WASHINGTON
  • EP2324352B1 patent drawingFigure 1a~1c
  • EP2324352B1 patent drawingFigure 1Aa~1Ac
  • EP2324352B1 patent drawingFigure 2a~2e

AI summary

Embodiments of the present invention relate to methods and apparatuses for the discretization and manipulation of sample volumes that is simple, robust, and versatile. It is a fluidic device that partitions a sample by exploiting the interplay between fluidic forces, interfacial tension, channel geometry, and the final stability of the formed droplet and/or discretized volume. These compartmentalized volumes allow for isolation of samples and partitioning into a localized array that can subsequently be manipulated and analyzed. The isolation of the discretized volumes along with the device's inherent portability render our invention versatile for use in many areas, including but not limited to PCR, digital PCR, biological assays for diagnostics and prognostics, cancer diagnosis and prognosis, high throughput screening, single molecule and single cell reactions or assays, the study crystallization and other statistical processes, protein crystallization, drug screening, environmental testing, and the coupling to a wide range of analytical detection techniques for biomedical assays and measurements. The minimal fluid interconnects and simple flow geometry makes the device easy to use and implement, economical to fabricate and operate, and robust in its operations.