Cell Cage Array for Parallel Single Cell Genetic Analysis

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Solution Overview

Problem

Current methods for genetic analysis of single cells are limited by the small amount of genetic material available, making it difficult to perform high-throughput, parallel nucleic acid analysis, and existing technologies are costly, time-consuming, and not straightforward for large-scale single-cell analysis.

Innovation Solution

A method involving a cell cage array with large and small openings to isolate single cells, followed by mounting on a microarray with unique DNA barcodes for nucleic acid amplification and sequencing, allowing for parallel analysis of nucleic acids from millions of cells using simple and inexpensive technology.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional single cell techniques are used, then genetic analysis can be performed on individual cells, but the process is slow, tedious, and limited to analyzing only a handful of cells at a time

Engineering Contradiction:
Improvenumber of cells analyzed per runVSAvoidtime required for analysis
Core Design Contradiction:
ProductivityVSLoss of time

Solution Approach 1:

The system segments the cell analysis process into individual cell isolation within separate cell cages, allowing parallel processing of thousands of cells simultaneously. Each cell cage acts as an independent reaction chamber, enabling massive parallelization of nucleic acid extraction and analysis.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The cell cage array serves as an intermediary structure that bridges single-cell isolation and high-throughput analysis. The array format allows cells to be captured individually while maintaining organization for parallel processing, converting a sequential process into a parallel one.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Productivity

If microfluidics or robotics are used to scale up single cell analysis, then more cells can be analyzed in parallel, but the equipment becomes costly and the process becomes complex

Engineering Contradiction:
Improvethroughput of cell analysisVSAvoidcomplexity of equipment and process
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The cell cage array serves multiple functions: cell isolation, nucleic acid extraction, and parallel analysis platform. This multi-functional design eliminates the need for separate complex microfluidic devices or robotic systems, achieving high throughput with simpler, more versatile equipment.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The system uses inexpensive, disposable cell cage arrays that can be discarded after a single use, eliminating the need for expensive, complex, and difficult-to-maintain microfluidic devices or robotic systems. The low cost allows for high throughput without proportionally increasing equipment investment.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

3Productivity

If microarrays with unique DNA barcodes are used, then parallel analysis of millions of cells becomes possible, but the system requires integration of multiple components and processes

Engineering Contradiction:
Improvescaling capacity to millions of cellsVSAvoidintegration complexity of system components
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The system merges cell cage array technology with microarray technology featuring unique DNA barcodes. This combination allows each cell to be individually isolated and simultaneously tagged with a unique identifier, enabling parallel tracking and analysis of millions of cells through a single integrated platform.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The system changes the scale parameter from analyzing handfuls of cells to millions of cells by implementing high-density cell cage arrays with unique DNA barcodes. This parameter change enables massive parallelization while the standardized protocol maintains simplicity despite the increased scale.

Inventive Principle:
Principle #35Parameter changes

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 massively parallel nucleic acid analysis of single cells, scaling to millions of cells with reduced costs and time, while maintaining high accuracy and reproducibility, overcoming the limitations of existing techniques.

Implementation Method 1

each cell cage comprises a large opening adjacent to said cell reservoir and at least one small opening adjacent to said enclosed reservoir, wherein the liquid medium flows through large openings of said cell cages, that are large enough for a single cell to enter each cage on the side adjacent to the cell reservoir, and through the small openings of said cell cages, that are small enough to prohibit cells to exit on the side adjacent to the enclosed reservoir

Methodology Applied
Scientific EffectPhysical containment through size-selective filtration: Filter (physical)

Implementation Method 2

mounting said cell cage array on a microarray comprising a plurality of DNA spots, wherein each DNA spot comprises a probe and a nucleic acid barcode unique for said DNA spot

Methodology Applied
Scientific EffectNucleic acid hybridization:

Implementation Method 3

introducing a buffer and inducing conditions for cell lysis and nucleic acid synthesis and amplification, to produce amplified nucleic acid molecules in the cages

Methodology Applied
Scientific EffectIn vitro transcription:

Data Source

PatentEP3253491B1System and method for single cell genetic analysis
Publication Date: 2021.03.24 TECHNION RES & DEV FOUND LTD
  • EP3253491B1 patent drawingFigure 1A~1C
  • EP3253491B1 patent drawingFigure 1D~1F
  • EP3253491B1 patent drawingFigure 1G~1H

AI summary

The present invention provides methods and systems for parallel analysis of a single cell's nucleic acid.