Drop-Carrier Particle Structures for Uniform Single-Cell Droplets
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Solution Overview
Problem
Current methods for single-molecule or single-cell assays face challenges in uniformly compartmentalizing small volumes without complex microfluidic instruments, and introducing solid supports or unique barcodes is difficult, leading to inefficiencies and high costs.
Innovation Solution
The development of sub-millimeter scale drop-carrier particles with well-defined 3D structures and chemical functionalities that create monodisperse droplet volumes, allowing for compartmentalization and association with solid supports, enabling reactions without microfluidics, and using sculpted microfluidic flows to fabricate Janus particles that stabilize aqueous droplets in an oil phase.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If microfluidic approaches are used to create monodisperse emulsions, then uniform droplet volumes are achieved, but high instrument cost and complexity are incurred
Solution Approach 1:
The patent uses disposable microwell arrays with hydrophobic coatings that can be easily discarded after use. This eliminates the need for expensive, complex microfluidic instruments while achieving uniform droplet volumes through the predefined well geometry and hydrophobic surface properties that prevent droplet coalescence.
Solution Approach 2:
The patent replaces the mechanical microfluidic pumping and flow control systems with a static microwell array system. Uniform droplet formation is achieved through the geometric constraints of the microwells and hydrophobic surface chemistry rather than through active mechanical control of fluid flow.
2Productivity
If microfluidic pumping systems are used, then droplet formation is achieved, but large dead-volumes limit small sample volume usage
Solution Approach 1:
The patent segments the sample into individual microwells, each containing a single droplet. This segmentation approach eliminates the need for large dead-volumes in pumping systems because the sample is partitioned into many small, discrete reaction compartments, allowing efficient use of minimal sample volumes.
Solution Approach 2:
The patent uses thin hydrophobic film coatings on the microwell surfaces to contain the droplets. This thin-film approach minimizes the volume of reagents and sample required compared to bulk microfluidic channels, enabling effective use of small sample volumes while maintaining droplet formation capability.
3Adaptability or versatility
If solid supports with reagents and barcodes are introduced into microwell arrays, then digital ELISA and single-cell RNAseq are enabled, but Poisson statistics limitations arise
Solution Approach 1:
The patent pre-loads solid supports with reagents and barcodes into the microwells before adding the sample. This preliminary action ensures that each compartment is properly prepared with the necessary components, enabling digital ELISA and single-cell RNAseq assays while maintaining uniform compartmentalization by avoiding post-loading variations.
Solution Approach 2:
The patent designs the system so that the solid supports themselves provide the necessary reagents and barcodes without requiring external intervention during the assay. This self-service approach reduces variability introduced by manual loading operations and maintains uniform compartmentalization while enabling versatile assay capabilities.
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 cost-effective, uniform compartmentalization of droplets compatible with standard equipment, facilitating single-molecule and single-cell assays with ease of use and compatibility with standard benchtop workflows, reducing the need for complex instruments.
Implementation Method 1
The interior region 14 of the drop-carrier particle 12 is, in one exemplary embodiment, hydrophilic. The exterior region 16 of the drop-carrier particle 12 is, in one exemplary embodiment, hydrophobic.
Implementation Method 2
sculpted microfluidic flows to fabricate Janus particles that stabilize aqueous droplets in an oil phase
Implementation Method 3
The combined drop-carrier particle with the dispersed phase (e.g., aqueous phase) contained therein is referred to as a particle-drop. The selective association results in compartmentalization of the dispersed phase solution into sub-microliter-sized volumes
Data Source
Figure 1
Figure 2A~2C
Figure 3A~4C
AI summary
The invention is directed to a particle system comprising a plurality of three-dimensional particles, each particle having an interior region defining a three-dimensional cavity or void and an exterior region, wherein the cavity or void is open to an external environment of the three-dimensional particle and wherein the cavity or void has a volume between about 1 pL and about 125 nL and a molecular capture region disposed on the interior region of the particles configured to capture cells, and to a method of performing an assay using the particle system.