Microfluidic Chamber Herringbone Structures for Cell Isolation

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Solution Overview

Problem

Current methods for isolating cells or virus particles of interest from biological fluids, particularly those present in low numbers amidst a high background of other cells or particles, face challenges in achieving high binding efficiency, specificity, and recovery, and often require labeled binding agents for identification.

Innovation Solution

A method involving labeling cells or virus particles with antibodies or antibody mimetics and contacting them with a solid support featuring an array of defined isolated spots of binding agents, combined with a microfluidic chamber with herringbone-structured transverse structures to facilitate chaotic mixing and enhance binding, allowing for direct identification and isolation without labeled agents.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If cells are isolated using conventional methods with labeled binding agents, then identification capability is improved, but device complexity and operational complexity increase

Engineering Contradiction:
Improveidentification capabilityVSAvoiddevice complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts the labeling step from the identification process. By using unlabeled binding agents for capture and relying on the intrinsic properties of cells (morphology, autofluorescence) for identification, the complex labeling and detection procedures are removed, simplifying the device while maintaining identification capability

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent enables cells to serve themselves for identification purposes. Instead of requiring external labels, the system utilizes the cells' own characteristics (shape, size, autofluorescence) to enable identification, thereby eliminating the need for additional labeling reagents and detection equipment

Inventive Principle:
Principle #25Self-service

2Measurement precision

If cells are isolated using conventional methods with labeled binding agents, then identification capability is improved, but ease of operation deteriorates

Engineering Contradiction:
Improveidentification capabilityVSAvoidease of operation
Core Design Contradiction:
Measurement precisionVSEase of operation

Solution Approach 1:

The patent removes the cumbersome labeling and detection steps from the operational workflow. By capturing cells with unlabeled binding agents and identifying them through intrinsic properties, the procedure becomes simpler and more straightforward to perform

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

Instead of labeling cells to enable identification, the patent inverts the approach by using unlabeled capture and relying on the cells' natural properties for identification, thereby simplifying the operational steps required

Inventive Principle:
Principle #13The other way round (Inversion)

3Productivity

If cells are captured on non-planar surfaces, then binding efficiency is improved, but accessibility for subsequent analysis deteriorates

Engineering Contradiction:
Improvebinding efficiencyVSAvoidaccessibility for subsequent analysis
Core Design Contradiction:
ProductivityVSEase of operation

Solution Approach 1:

The patent applies local quality by creating micropatterned arrays of binding agents on an otherwise planar surface. The binding activity is localized to specific micropatterned regions, maintaining high binding efficiency at these locations while the overall planar surface remains accessible for downstream operations

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The patent transitions from three-dimensional non-planar capture surfaces to two-dimensional micropatterned arrays on a planar substrate. This dimensional reduction maintains binding capacity while dramatically improving accessibility for subsequent single-cell analysis operations

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

4Quantity of substance

If cells are isolated from large volumes of biological fluid, then recovery of rare cells is improved, but binding efficiency deteriorates due to low cell concentration

Engineering Contradiction:
Improverecovery of rare cellsVSAvoidbinding efficiency
Core Design Contradiction:
Quantity of substanceVSProductivity

Solution Approach 1:

The patent segments the large volume of biological fluid into multiple smaller aliquots that are processed sequentially through the microfluidic device. This segmentation allows thorough mixing and binding in each small volume while maintaining the ability to process large total volumes, thereby improving both recovery and binding efficiency

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent incorporates preliminary chaotic mixing actions within the microfluidic device before the cell capture step. This preliminary mixing ensures that rare cells are evenly distributed and have maximum opportunity to encounter binding agents, improving binding efficiency even when processing large volumes with low cell concentrations

Inventive Principle:
Principle #10Preliminary action

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 high specificity and recovery of cells or virus particles of interest, facilitating their immobilization and subsequent single-cell analysis on a plane surface, improving the efficiency of cell capture and reducing false positives.

Implementation Method 1

a surface opposing the solid support, said surface having transverse structures that facilitate chaotic mixing of the fluid containing the cells or virus particles of interest when said fluid flows along said surface

Methodology Applied
Scientific EffectChaotic mixing: Convection

Implementation Method 2

contacting the labeled cells or virus particles of interest with a solid support (biochip), said solid support comprising an array of defined isolated spots of a solid support-bound second binding agent, wherein the first and second binding agents can bind to each other

Methodology Applied
Scientific EffectSpecific binding: Adsorption

Data Source

PatentEP3256572B1Immobilization of cells or virus particles on protein structures using a microfluidic chamber
Publication Date: 2019.08.21 KARLSRUHER INST FUR TECH
  • EP3256572B1 patent drawingFigure 1
  • EP3256572B1 patent drawingFigure 2
  • EP3256572B1 patent drawingFigure 3a~3d

AI summary

The present invention relates to methods for the immobilization of cells or virus particles of interest expressing one or more predetermined surface marker(s) on defined spots on a solid support, comprising the steps of providing a biological fluid sample suspected of containing cells or virus particles of interest; labeling the cells or virus particles of interest with antibodies, antibody fragments or antibody mimetics directed against the one or more predetermined surface marker(s) and carrying a first binding agent; and contacting the labeled cells or virus particles of interest with a solid support (biochip), said solid support comprising an array of defined isolated spots of a solid support-bound second binding agent, wherein the first and second binding agents can bind to each other. The present invention further relates to devices for the isolation of cells or virus particles of interest expressing one or more predetermined surface marker(s), comprising a microfluidic chamber comprising a solid support comprising an array of defined isolated spots of a solid support-bound binding agent, and a surface opposing the solid support, said surface having transverse structures that facilitate chaotic mixing of the fluid containing the cells or virus particles of interest when said fluid flows along said surface; and a capillary micropipette.