Single-Particle Cell Analysis With Aseptic Micro-Flow Cartridge Sorting
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Solution Overview
Problem
Existing cell sorting and analysis methods for regenerative medicine face challenges such as aseptic sorting without damage, adjusting sample concentration, handling large or clumped cells, purifying differentiating cells, and analyzing single cells, particularly in the context of water-in-oil emulsion droplets, with issues like contamination, damage, and inefficient sorting.
Innovation Solution
A cell analysis apparatus and method utilizing a flow path cartridge with sealed reservoirs, controlled air pressure, and deformable membranes for aseptic sorting, combined with fluorescence detection and adjustable flow rates, enables sorting and analyzing cells and clumps in an enclosed space, ensuring minimal damage and efficient removal of undifferentiated cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional cell sorters are used for sorting cells in regenerative medicine, then cells can be sorted and concentrated, but cells are subjected to heavy damage and the process is not aseptic
Solution Approach 1:
The patent employs a disposable micro-flow path cartridge that is used once and then discarded, eliminating the need for complex sterilization procedures and preventing cross-contamination. This single-use approach maintains aseptic conditions while enabling efficient cell sorting without the heavy damage caused by conventional reusable sorters.
Solution Approach 2:
The system separates the sorting function into a dedicated micro-flow path cartridge that can be independently replaced. This segmentation allows the main sorting apparatus to remain sterile and reusable while the disposable cartridge handles the actual cell processing, resolving the contradiction between productivity and contamination risk.
2Measurement precision
If flow cytometry is used to identify cells, then various types of cells can be detected, but the method is not suitable for regenerative medicine due to lack of asepsis and cell damage
Solution Approach 1:
The micro-flow path cartridge is designed as a disposable component that maintains aseptic conditions throughout the cell identification and sorting process. The cartridge is sterilized during manufacturing and used once, preventing contamination while enabling flow cytometry-based cell identification without the harmful effects of conventional reusable systems.
3Quantity of substance
If magnetic particles are used to separate cells, then specific cells can be isolated, but the method does not achieve complete removal of undifferentiated cells
Solution Approach 1:
The patent combines multiple separation mechanisms within the micro-flow path cartridge, including magnetic particle-based separation and flow-based sorting. This merged approach achieves complete removal of undifferentiated cells by utilizing the complementary strengths of different separation methods in an integrated system.
Solution Approach 2:
The system incorporates detection units that monitor the cell population in real-time, providing feedback to the sorting mechanism. This allows the system to adjust separation parameters dynamically to ensure complete removal of undifferentiated cells, achieving purification completeness that single-method approaches cannot attain.
4Productivity
If conventional sorting methods are used, then cells can be sorted, but the process is complex and not suitable for practical regenerative medicine applications
Solution Approach 1:
The disposable micro-flow path cartridge integrates multiple functions (flow control, sorting, detection) into a single pre-configured unit. This eliminates the need for complex external control systems and sterilization procedures, simplifying the overall system while maintaining high sorting capability for practical regenerative medicine applications.
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 allows for aseptic sorting of large cell clumps and efficient removal of undifferentiated cells, achieving zero undifferentiated cells per 10^6 differentiating cells, while enabling detailed gene analysis of individual cells or clumps, suitable for regenerative medicine applications.
Implementation Method 1
When a cell passes through an illumination region, light is scattered depending on the size, shape, and refractive index of the cell
Implementation Method 2
To detect a cell specifically dyed with a fluorescent dye by fluorescence, the wavelength of the laser beam is determined in accordance with the type of the fluorescent dye
Implementation Method 3
a third-A reservoir connected to the fourth branched flow path for delivering a pulse flow thereto; a third-B reservoir connected to the fifth branched flow path for changing a particle course through the pulse flow
Data Source
AI summary
An object of the present invention is to purify and concentrate differentiating cells derived from ES cells, iPS cells, or the like without damaging them.The above problem can be solved by an apparatus for analyzing and separating particles comprising: a flow path cartridge, an illumination unit, a detection unit for detecting particles of interest, a force generating unit,wherein a sample liquid reservoir (sample reservoir) connected to a first flow path; a fourth branched flow path and a fifth branched flow path which are connected to the first flow path; a third-A reservoir connected to the fourth branched flow path; a third-B reservoir connected to the fifth branched flow path; and a fourth reservoir for reserving particles which are not sorting; are formed on the cartridge, and each reservoir comprise a means which equalizes an air pressure in the each reservoir with an air pressure of an in-device air pressure control system, and a stream of the flow path in the cartridge is controlled by controlling the air pressure in the each reservoir through the each in-device air pressure control system.