Repeated Particle Sorting for High-Purity Cell and Droplet Handling
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Solution Overview
Problem
Conventional cell sorting and dispensing technologies face challenges such as high cell damage, low throughput, contamination, limited droplet size, and difficulty in handling water-in-oil droplets, especially in fluorocarbon oil, and inadequate analysis of cell clusters.
Innovation Solution
A method and apparatus for purifying target particles by repeated sorting, dispensing single particles using a flow-path chip with controlled air pressure, and analyzing cell clusters based on scattered light signal ratios.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If conventional jet in air type cell sorter is used to separate target cells from high concentration non-target cells, then sorting speed is fast, but cell damage is high and processing time is excessively long for high concentration samples
Solution Approach 1:
The sorting process is divided into multiple sequential stages using a series of sorting chambers with decreasing aperture sizes. Target cells are sorted in each chamber according to their size characteristics, progressively separating them from non-target cells. This segmentation allows high-speed sorting without the need for excessive dilution, reducing processing time while maintaining cell integrity.
Solution Approach 2:
The invention replaces the conventional jet in air mechanical sorting system with a microfluidic-based sorting system. This substitution uses controlled fluid flow and pressure differential mechanisms to achieve cell separation, eliminating the high-impact jet mechanism that causes cell damage while maintaining efficient sorting speed.
2Speed
If conventional jet in air type cell sorter is used for high concentration samples, then sorting speed is maintained, but processing time becomes excessively long requiring sample dilution
Solution Approach 1:
The sorting process is divided into multiple sequential stages using a series of sorting chambers with decreasing aperture sizes. Target cells are sorted in each chamber according to their size characteristics, progressively separating them from non-target cells. This segmentation allows high-speed sorting without the need for excessive dilution, reducing processing time while maintaining cell integrity.
Solution Approach 2:
The microfluidic system maintains continuous flow and sorting action through multiple chambers without interruption. The airtight connection between chambers ensures continuous pressure differential-driven flow, allowing the system to process high concentration samples continuously at high speed without the time loss associated with dilution and reprocessing.
3Manufacturing precision
If repeated sorting is performed to purify target particles from high concentration non-target particles, then purity is improved, but processing complexity increases
Solution Approach 1:
The sorting system is segmented into multiple independent chambers connected in series, each with progressively smaller apertures. This physical segmentation allows repeated sorting to occur automatically as particles flow through each chamber, achieving high purification precision without requiring complex external control mechanisms for repeated processing.
Solution Approach 2:
Each sorting chamber serves multiple functions: it acts as both a filtering stage and a sorting stage, and the same chamber structure is reused across all stages with varying aperture sizes. This universality reduces device complexity by using a standardized modular design that performs multiple sorting functions.
4Ease of manufacture
If water in oil droplets are handled in fluorocarbon oil using conventional methods, then droplet formation is achieved, but dispensing is difficult due to floating and settling issues
Solution Approach 1:
The invention uses pressure differential control through airtight connections between chambers to manipulate droplet movement and dispensing. By controlling the pressure in each chamber, the system can precisely control the movement of water-in-oil droplets through the sorting path and into collection vessels, overcoming the floating and settling issues that plague conventional dispensing methods.
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 rapid purification of target particles from high-concentration samples, reliable dispensing of single particles and water-in-oil droplets, and accurate determination of cell clusters, reducing processing time and improving purity.
Implementation Method 1
a flow cytometer, and a method for analyzing cell cluster by analyzing flow cytometry data
Data Source
AI summary
One object of the present invention is to provide a method or apparatus for purifying target particles from high-concentration particles in a short time. The above problem can be solved by a method for purifying target particles, characterized in that the method comprises a step of sorting the target particles from a high concentration of non-target particles, wherein the sorting step is repeated for three times or more.