Microchannel Cassette Sorting for Viable High-Throughput Cell Isolation
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Solution Overview
Problem
Current methods for identifying, isolating, and characterizing biological components such as cells and proteins are time-consuming, prone to sample contamination, and inefficient in detecting multiple positive signals, isolating viable cells, and differentiating single cells from multiple cells, with limitations in the number of cells that can be screened effectively.
Innovation Solution
A cassette system with microchannels and a housing structure that allows for the detection and sorting of target particles, including viable cellular material, using electromagnetic radiation and a method that involves loading a sample mixture into microchannels, scanning for target particles, and extracting them using an extraction beam, with the ability to process a high volume of cells per second while maintaining cell viability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used for identifying and isolating biological components, then the process can be completed with standard equipment, but the process becomes time-consuming and inefficient
Solution Approach 1:
The device segments the sample into individual cells using microfluidic channels, allowing parallel processing of multiple cells simultaneously. This segmentation enables high-throughput screening by distributing cells across numerous channels that can be analyzed concurrently, dramatically increasing productivity while reducing total sorting time.
Solution Approach 2:
The patent replaces conventional mechanical sorting mechanisms with optical detection and electromagnetic radiation-based sorting. Flow cytometry and laser-based identification systems substitute for mechanical separation methods, enabling faster non-contact cell manipulation and sorting, thereby improving throughput and reducing processing time.
2Manufacturing precision
If multiple selection steps are used to isolate target cells, then the purity of isolated cells can be improved, but the process becomes more complex and time-consuming
Solution Approach 1:
The device merges multiple selection functions into a single integrated platform that combines microfluidic cell separation, optical detection, and laser-based sorting. This consolidation maintains high purity requirements while reducing the number of discrete steps needed, as all selection criteria can be applied simultaneously through multi-parameter flow cytometry and coordinated laser activation.
Solution Approach 2:
The sorting device is designed with multi-functionality, capable of performing various cell selection operations through programmable laser activation and multiple detection channels. A single device can handle different cell types, markers, and selection criteria without requiring separate specialized equipment for each function, thereby maintaining precision while reducing overall system complexity.
3Measurement precision
If conventional detection methods are used, then the equipment required is simple, but the ability to detect multiple positive signals and differentiate single cells from multiple cells is insufficient
Solution Approach 1:
The patent employs optical detection systems including flow cytometry and laser-based fluorescence detection to replace simple visual or mechanical detection methods. These optical systems can simultaneously detect multiple fluorescent markers and distinguish single cells from cell clusters based on light scattering patterns and fluorescence intensity, achieving high measurement precision through non-mechanical optical fields.
4Productivity
If high-speed cell sorting is implemented, then the number of cells screened per second increases, but cell viability may be compromised
Solution Approach 1:
The device uses laser-based optical sorting instead of mechanical contact methods to manipulate and sort cells at high speeds. The laser creates localized electromagnetic fields that can deflect or manipulate individual cells without physical contact, minimizing mechanical stress and maintaining cell viability even during high-throughput sorting operations exceeding 100,000 cells per second.
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, efficient, and sterile sorting of target particles with high purity and viability, suitable for pharmaceutical compositions, particularly hematopoietic stem cells, with the capability to screen and extract cells at rates exceeding 100,000 cells per second and achieve greater than 95% purity and viability.
Implementation Method 1
a transmissive portion located in one or each of the first housing and the second housing, wherein the transmissive portion permits transmission of electromagnetic radiation from outside of the cassette to the substrate
Implementation Method 2
an excitation light source to emit an excitation beam to generate fluorescence light from target particles
Implementation Method 3
an extraction laser to provide an extraction beam to remove target particles from the surface or a plurality of channels
Data Source
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AI summary
This disclosure provides methods and apparatuses for sorting target particles. In various embodiments, the disclosure provides a cassette for sorting target particles, methods for sorting target particles, methods of loading a microchannel for maintaining sample material viability, methods of quantifying sample material, and an optical apparatus for laser scanning and particle sorting.