Inert Gas Bubble Segmentation for Hydrodynamic Particle Sorting

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Solution Overview

Problem

Current flow cytometry methods face challenges in performing fast and continuous analysis of individual cells or cell populations, especially when cells need to be digested or their membrane destroyed, as the cells tend to mix and lose their distinct identity, leading to contamination and interference in measurement techniques.

Innovation Solution

The introduction of inert gas bubbles between hydrodynamically focused particles in a channel allows for sharp and contaminant-free separation of cells, regulating the gas introduction to synchronize hydrodynamic focusing, sample preparation, and detection, enabling individual cell analysis without the need for fluorescence activation or labeling.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If cells are digested or membrane destroyed for complex analysis, then analytical capability is improved, but cell separation and identification capability deteriorates due to mixing and loss of distinct identity

Engineering Contradiction:
Improveanalytical capabilityVSAvoidcell separation and identification
Core Design Contradiction:
Adaptability or versatilityVSReliability

Solution Approach 1:

The continuous liquid flow carrying multiple cells is segmented into discrete portions by introducing gas bubbles between hydrodynamically focused cells. This segmentation isolates each cell in its own liquid portion, preventing mixing even when cells are digested or membranes are destroyed, thus maintaining both analytical capability and cell identification reliability

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Gas bubbles are introduced as an intermediary substance between adjacent hydrodynamically focused cells. These bubbles act as physical barriers that prevent direct contact and mixing between cell-containing liquid portions, enabling reliable cell separation and identification while allowing complex analytical processing

Inventive Principle:
Principle #24Intermediary (Mediator)

2Adaptability or versatility

If fluorescence labeling is used for cell differentiation, then cell differentiation capability is improved, but measurement accuracy deteriorates due to spectral interferences limiting parameter examination

Engineering Contradiction:
Improvecell differentiation capabilityVSAvoidparameter examination accuracy
Core Design Contradiction:
Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The invention extracts and eliminates the need for fluorescence labeling by directly introducing gas bubbles for cell separation. This removes the source of spectral interferences while maintaining cell differentiation capability through alternative means, thereby improving measurement precision without sacrificing versatility

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The optical/chemical fluorescence labeling system is replaced with a purely mechanical separation approach using gas bubbles. This substitution eliminates spectral interferences entirely while achieving cell differentiation through physical separation and individual analysis of each cell

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

3Productivity

If continuous flow analysis is performed on cells in liquid, then analysis speed is improved, but sample preparation quality deteriorates leading to contamination and interference

Engineering Contradiction:
Improveanalysis speedVSAvoidsample preparation quality
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

Gas bubbles are introduced periodically between hydrodynamically focused cells to create discrete, separated portions of cell-containing liquid. This periodic action maintains continuous flow for high-speed analysis while ensuring each cell portion is isolated and contamination-free, thus preserving both productivity and sample preparation quality

Inventive Principle:
Principle #19Periodic action

Solution Approach 2:

The gas bubble introduction creates local isolation zones around individual cells or small cell groups. Each localized portion maintains high preparation quality through physical separation, while the overall system continues to operate in continuous flow mode for rapid analysis

Inventive Principle:
Principle #3Local quality

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 method provides accurate, label-free separation and analysis of cells, preventing contamination and interference, allowing for the determination of complex analytes like biomolecules without altering the sample, and enabling the analysis of a wide range of parameters without spectral interferences.

Implementation Method 1

separating or sorting hydrodynamically focused particles contained in a liquid within a channel

Methodology Applied
Scientific EffectHydrodynamic focusing:

Implementation Method 2

In a channel with wall, inert gas bubbles are introduced between hydrodynamically focused particles

Methodology Applied
Scientific EffectGas bubble separation: Bubble

Data Source

PatentEP4394350A1Apparatus and method for separating particles
Publication Date: 2024.07.03 KROMAT
  • EP4394350A1 patent drawingFigure 1a~1b
  • EP4394350A1 patent drawingFigure 2a~2b
  • EP4394350A1 patent drawingFigure 3

AI summary

The invention relates to the field of analysis of membrane-bound compartments or analytes therein. The present invention relates to an apparatus for separating or sorting hydrodynamically focused particles contained in a liquid said apparatus comprising - a channel with wall, for conveying a liquid comprising hydrodynamically focused particles; - gas inlet opening for introducing inert gas bubbles into the flow channel to separate groups of hydrodynamically focused particles of defined or pre-determined particle number; - means for regulation of the introduction of inert gas into the flow channel; and - preferably means for counting the particles The present invention further relates to a method for sorting the hydrodynamically focused particles in the flow channel using the apparatus of the invention.