Shaped Particle Nanovials for Single-Cell Flow Assay Isolation
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Solution Overview
Problem
Existing biological assay technologies are limited in their ability to perform functional analysis and sorting of individual cells at high-throughput due to volume constraints and cross-talk issues, requiring specialized and expensive equipment, which hinders widespread adoption.
Innovation Solution
The use of shaped particles or 'nanovials' that act as suspendable and sortable microwells, allowing for sub-nanoliter volume containment of single cells, enabling fluid exchange and molecular readouts, and compatible with commercial flow cytometers for high-throughput analysis and sorting.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If microwell plate format is used for biological assays, then experiments can be scaled and integrated with lab automation infrastructure, but the volume is hundreds of thousands of times larger than single cells limiting functional property analysis
Solution Approach 1:
The invention segments the microwell plate into individual single-cell compartments using shaped particles. Each particle contains a single cell in a minimal volume cavity, effectively dividing the large microwell volume into numerous tiny isolated compartments. This allows high-throughput analysis (hundreds of thousands of cells) while reducing each cell's volume from microliters to attoliters, enabling functional property analysis at single-cell resolution.
2Ease of operation
If standard microwell plates are used, then fluid exchange is simple, but cross-talk between samples occurs and prevents single-cell resolution
Solution Approach 1:
The shaped particles act as intermediary containers between the bulk fluid environment and individual cells. Each particle's cavity physically isolates the cell and its secretions from surrounding samples, preventing cross-talk. The particles can be manipulated as a suspension in standard microwell plates, maintaining ease of fluid exchange while the particle cavities provide the necessary physical barriers to prevent sample contamination.
3Measurement precision
If microfluidic technologies are used to create cell-isolating volumes, then single-cell analysis based on secreted molecules is enabled, but the ability to add reagents and wash reactions is limited
Solution Approach 1:
The shaped particles provide universal functionality that combines the advantages of microfluidic isolation with microwell plate versatility. The particles can be used in standard microwell plates, allowing integration with existing lab automation infrastructure. Reagents can be added to the bulk fluid to interact with cells in particle cavities, and wash reactions can be performed by fluid exchange in the plate. The particles themselves can be functionalized with capture agents, making the system adaptable to various assay types while maintaining single-cell resolution.
4Volume of stationary object
If microfluidic devices are used, then volumes approaching cell size are achieved, but specialized and expensive equipment is required hindering widespread adoption
Solution Approach 1:
Instead of using complex microfluidic devices to create isolation volumes, the invention uses simple shaped particles (spheres, beads, or other geometric forms) that can be mass-produced using conventional manufacturing methods. These particles are suspended in standard microwell plates, copying the isolation function of microfluidic chambers but using inexpensive, easily manufactured components. This eliminates the need for specialized microfluidic equipment while achieving the same single-cell volume isolation, making the technology accessible to widespread adoption.
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
Enables high-throughput screening of >500,000 events per hour, identifying rare antibody-secreting cells within a day, and is compatible with standard assay formats, overcoming volume limitations and cross-talk issues.
Implementation Method 1
Fluids are easily exchanged by centrifugation and pipetting
Implementation Method 2
sorting of individual cells based on light scatter properties
Implementation Method 3
labeled with fluorescent reporters enabling sorting based on secreted molecules
Data Source
AI summary
Suspendable shaped particles or nanovials are used to viably sort single cells based on their secreted product at high-throughput using only commonly accessible lab infrastructure. These shaped particles act as a solid support which facilitates cell attachment, templates formation of uniform aqueous compartments which prevent cross-talk between cells, and captures secreted proteins. Using this platform, high-throughput screening of producer cells on relative IgG production, B/plasma cells based on secreted antibody binding to antigen, and T cells based on cytokine secretion is demonstrated using commercially available flow sorters. These shaped particles are easily distributed and used, democratizing access to high-throughput functional cell screening.