Microwell Array with Unique DNA Tags for High-Throughput Cell Screening

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Solution Overview

Problem

Traditional methods for cultivating and screening biological entities are slow, laborious, and expensive, often failing to culture microbes and requiring complex methods for identifying metabolites, enzymes, or therapeutic benefits, which limits the discovery of new insights and products.

Innovation Solution

A microfabricated device with a high density array of microwells, where each well contains a unique tag for identifying the species cultivated, allowing for high-throughput cultivation, screening, and localization of cells using nucleic acid molecules with target-specific and location-specific sequences, enabling efficient cultivation, analysis, and product development.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If traditional cultivation methods are used, then simplicity of operation is maintained, but productivity and throughput are low

Engineering Contradiction:
Improvethroughput of cultivation and screeningVSAvoidcomplexity of cultivation system
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The system segments the cultivation process into individual microwells (e.g., 1000 microwells per slide), allowing parallel cultivation of many biological entities simultaneously. Each microwell acts as an independent cultivation unit, enabling high-throughput processing while maintaining simple individual well operations.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The microwell array system serves multiple functions: cultivation, screening, identification, and localization of biological entities. The same platform handles diverse tasks including growing cells, screening for metabolites/enzymes, and identifying species through unique tags, eliminating the need for separate specialized equipment for each function.

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

2Measurement precision

If traditional screening methods are used, then method simplicity is maintained, but measurement precision and identification accuracy are insufficient

Engineering Contradiction:
Improveaccuracy of species identificationVSAvoidcomplexity of screening method
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

Unique tags (DNA barcodes, fluorescent markers, or magnetic beads) serve as intermediaries between the biological entities and the identification system. These tags provide precise species identification without requiring complex direct analysis of the biological entities themselves, simplifying the screening process while maintaining high accuracy.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The system uses fluorescently labeled unique tags that emit different colors or fluorescence intensities corresponding to different species or conditions. This optical signaling enables rapid, automated, and precise identification of biological entities through simple fluorescence detection, avoiding complex biochemical analysis.

Inventive Principle:
Principle #32Color changes

3Productivity

If high density microwell arrays are used, then productivity and throughput are improved, but ease of operation and handling become difficult

Engineering Contradiction:
Improvenumber of simultaneous cultivationsVSAvoidhandling of microwell arrays
Core Design Contradiction:
ProductivityVSEase of operation

Solution Approach 1:

The system combines thousands of microwells into a single integrated slide or chip that can be handled as one unit. This merging allows high-throughput cultivation while maintaining simple operation, as the entire array can be loaded, incubated, and analyzed together rather than handling individual wells separately.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The system replaces manual mechanical handling of individual wells with automated liquid handling robots and imaging systems. These automated systems perform pipetting, incubation, and analysis of the microwell arrays, eliminating the labor-intensive manual operations while maintaining ease of use through simple sample loading.

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

4Measurement precision

If unique tags with nucleic acid molecules are used, then measurement precision and identification accuracy are improved, but loss of substance and cost increase

Engineering Contradiction:
Improveaccuracy of location identificationVSAvoidconsumption of nucleic acid tags
Core Design Contradiction:
Measurement precisionVSLoss of substance

Solution Approach 1:

The system uses DNA barcodes as information copies that can be amplified through PCR. Instead of needing unique physical tags for each biological entity, the system uses replicable genetic sequences that can be copied and detected, dramatically reducing material consumption while maintaining precise identification capability.

Inventive Principle:
Principle #26Copying

Solution Approach 2:

The system changes the detection parameter from detecting rare unique molecules to detecting amplified nucleic acid sequences. By using PCR amplification, trace amounts of nucleic acid tags can be detected after exponential amplification, reducing the initial amount of substance needed while maintaining high detection sensitivity and precision.

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

This approach streamlines the cultivation workflow, supports high-throughput screening, and facilitates the identification and localization of biological entities, leading to the discovery of new insights and products, such as antibiotics and fertilizers, by efficiently culturing and analyzing cells in a high-density array format.

Implementation Method 1

the nucleic acid molecule comprising a target-specific nucleotide sequence for annealing to a target nucleic acid fragment present in at least one species of interest

Methodology Applied
Scientific EffectAnnealing:

Data Source

PatentEP3285926B1Kit and method for high throughput microbiology applications
Publication Date: 2022.03.02 ISOLATION BIO INC
  • EP3285926B1 patent drawingFigure 1
  • EP3285926B1 patent drawingFigure 2A~2C
  • EP3285926B1 patent drawingFigure 3A~3B

AI summary

A microfabricated device defining a high density array of microwells is described for cultivating cells from a sample. A series of unique tags may be disposed in the microwells to identify one or more species of cells and locate the particular microwells in which each species was cultivated. A unique tag may be a nucleic acid molecule including a target-specific nucleotide sequence for annealing to a target nucleic acid fragment and a location-specific nucleotide sequence predetermined to identify one or more microwells. The device may be incubated to grow a plurality of cells, which may be split into an analysis portion and a reserve portion. High throughput methods are described for cultivating, screening, and determining a relative and/or absolute abundance of cells from a sample.