Micro-fluidic Device Isolating Nucleic Acids via DEP Electrodes

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Solution Overview

Problem

Current methods for extracting nucleic acid materials from biological cells are inefficient in selectively capturing specific types of nucleic acids and often result in mixing of biological cells or capture objects, leading to contamination and reduced specificity.

Innovation Solution

A micro-fluidic device with dielectrophoresis (DEP) electrodes and isolation pens that allow for selective activation and deactivation of electrodes, enabling the placement, lysis, and capture of nucleic acid materials within individual cells, using capture objects with high specificity for particular types of nucleic acids, while preventing mixing between pens.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional nucleic acid extraction methods are used, then nucleic acid materials can be extracted from biological cells, but the methods lack efficiency in selectively capturing specific types of nucleic acids and result in mixing of biological cells or capture objects leading to contamination

Engineering Contradiction:
Improvespecificity of nucleic acid captureVSAvoidcontamination from mixing
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The device segments individual biological cells into separate isolation pens within the micro-fluidic channel, preventing mixing between cells. Each isolation pen acts as an independent compartment that maintains physical separation while allowing controlled medium exchange through diffusion, thereby eliminating cross-contamination and enabling specific capture of nucleic acids from individual cells.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent implements local quality by providing each isolation pen with specific capture objects tailored to capture particular types of nucleic acids (e.g., mRNA, miRNA, DNA) from individual cells. This localized specificity ensures that each pen performs a specialized function for capturing desired nucleic acid materials while maintaining physical isolation from other pens, thus improving both specificity and preventing contamination.

Inventive Principle:
Principle #3Local quality

2Measurement precision

If individual cells are processed separately to prevent contamination, then specificity is improved, but the complexity of the device increases due to multiple isolation pens and selective electrode activation

Engineering Contradiction:
Improvespecificity of nucleic acid captureVSAvoidmicro-fluidic device structure
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The micro-fluidic device employs a universal control mechanism where a single set of DEP electrodes can selectively activate and deactivate multiple isolation pens through patterned electrical fields. This multi-functional capability allows the system to process multiple individual cells in parallel while using a unified electrode structure, thereby reducing overall device complexity compared to having separate control mechanisms for each pen.

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

Solution Approach 2:

The patent utilizes micro-fluidic hydraulic principles to enable automatic medium exchange and cell manipulation through the isolation pens. By designing the channel geometry and opening positions to leverage fluid flow dynamics and diffusion, the system achieves automated cell processing and medium replacement without requiring complex mechanical actuators or additional control components for each individual pen.

Inventive Principle:
Principle #29Pneumatics and hydraulics

3Productivity

If capture objects with high specificity are used, then nucleic acid capture efficiency is improved, but the difficulty of preventing mixing between pens increases

Engineering Contradiction:
Improvenucleic acid capture efficiencyVSAvoidprevention of mixing
Core Design Contradiction:
ProductivityVSDifficulty of detecting and measuring

Solution Approach 1:

The isolation pens are nested within the micro-fluidic channel structure, with each pen containing capture objects that are themselves nested within the pen's interior space. This nested configuration ensures that capture objects with high specificity remain physically confined within their respective pens, preventing mixing between different pens while maintaining the high capture efficiency provided by the specific capture objects.

Inventive Principle:
Principle #7Nested doll (Nesting)

Solution Approach 2:

The patent introduces a liquid medium as an intermediary that facilitates controlled exchange between the isolation pens and the external environment. The medium allows for diffusion-based nutrient and reagent exchange while the physical barriers of the pen walls prevent direct mixing between pens. This intermediary mechanism enables high-specificity capture objects to function effectively without direct exposure to other pens' contents.

Inventive Principle:
Principle #24Intermediary (Mediator)

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 solution effectively captures specific nucleic acid materials with high specificity, preventing contamination and allowing for the correlation of captured nucleic acids with their originating cells, enhancing the efficiency and accuracy of nucleic acid extraction.

Implementation Method 1

an electrode activation substrate comprising dielectrophoresis (DEP) electrodes at a surface of said substrate, wherein each said electrode is configured to be selectively activated and deactivated

Methodology Applied
Scientific EffectDielectrophoresis: Dielectric Permittivity

Implementation Method 2

a single opening sized and positioned to permit exchange of a first liquid medium in said pen with a second liquid medium flowing past said opening in said channel by diffusion

Methodology Applied
Scientific EffectDiffusion: Diffusion

Data Source

PatentEP3083980B1Capturing specific nucleic acid materials from individual biological cells in a micro-fluidic device
Publication Date: 2021.10.13 BERKELEY LIGHTS INC
  • EP3083980B1 patent drawingFigure 1
  • EP3083980B1 patent drawingFigure 2A
  • EP3083980B1 patent drawingFigure 2B

AI summary

Individual biological cells can be selected in a micro-fluidic device and moved into isolation pens in the device. The cells can then be lysed in the pens, releasing nucleic acid material, which can be captured by one or more capture objects in the pens. The capture objects with the captured nucleic acid material can then be removed from the pens. The capture objects can include unique identifiers, allowing each capture object to be correlated to the individual cell from which the nucleic acid material captured by the object originated.