Laser Fluorescence Particle Sorting for Viable Cell Isolation
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Solution Overview
Problem
Current methods for identifying, isolating, and characterizing biological components, such as cells and proteins, are inefficient, often requiring multiple time-consuming steps and failing to prevent sample contamination, accurately detect multiple positive signals, isolate viable cells, or differentiate single cells from multiple cells, and are limited in throughput.
Innovation Solution
A system comprising an excitation light source, a detector, and an extraction laser with a scanner and circuitry to generate and detect fluorescence, allowing for the rapid sorting of target particles, including viable cells, by emitting an excitation beam to generate fluorescence and an extraction beam to remove particles from a surface or channels, with processing rates ranging from 5,000 to 100,000,000 particles per second.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional methods are used for identifying and isolating biological components, then sample contamination is prevented through multiple selection steps, but the process becomes time-consuming and reduces productivity
Solution Approach 1:
The device divides the sample into individual cells using microfluidic channels and acoustic fields, allowing each cell to be processed independently. This segmentation enables parallel processing of multiple cells simultaneously, increasing throughput while maintaining contamination prevention through physical isolation of each cell in its own channel.
Solution Approach 2:
The patent replaces conventional mechanical manipulation and multiple manual selection steps with acoustic fields for cell separation and laser-based detection for identification. This substitution automates the process, eliminating time-consuming manual operations while maintaining reliability through precise acoustic positioning and optical detection.
2Measurement precision
If multiple selection steps are implemented to ensure accurate detection, then detection precision improves, but device complexity and processing time increase
Solution Approach 1:
The device combines multiple detection functions into a single integrated platform: acoustic cell separation, fluorescent labeling detection, viability assessment, and isolation all occur in one continuous flow-through process. This merging eliminates the need for multiple separate selection steps while maintaining comprehensive detection accuracy through simultaneous multi-parameter analysis.
Solution Approach 2:
The microfluidic device performs multiple functions within a single system: cell sorting, fluorescent detection, viability assessment, and sample isolation. This multi-functionality allows the device to replace several separate apparatuses and selection steps, reducing overall device complexity while maintaining comprehensive detection capabilities.
3Productivity
If high throughput processing is achieved, then productivity increases, but the ability to accurately detect multiple positive signals and differentiate single cells from multiple cells decreases
Solution Approach 1:
By isolating each cell in its own microfluidic channel, the device ensures that fluorescent signals originate from single cells rather than aggregates. This physical segmentation enables accurate detection of multiple positive signals within individual cells while maintaining high throughput through parallel processing of many channels simultaneously.
Solution Approach 2:
The device incorporates real-time fluorescent detection that provides feedback on cell characteristics as they pass through the channels. This feedback mechanism allows the system to accurately identify multiple positive signals within cells and differentiate single cells from aggregates, with the ability to adjust isolation parameters dynamically based on detected signals.
4Productivity
If rapid cell isolation is performed, then processing speed increases, but cell viability may be compromised
Solution Approach 1:
The device replaces mechanical manipulation and physical handling of cells with acoustic fields for positioning and laser-based detection for identification. This substitution enables rapid isolation without mechanical stress, maintaining cell viability through contactless acoustic manipulation and precise laser targeting that minimizes thermal damage.
Solution Approach 2:
The system uses acoustic parameters (frequency, amplitude) to manipulate cell position and isolation timing, allowing rapid cell capture without mechanical force. By adjusting acoustic field parameters dynamically, the device achieves fast isolation rates while maintaining gentle handling conditions that preserve cell viability.
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 efficient and rapid sorting of target particles with high purity and viability, suitable for pharmaceutical compositions, achieving processing rates and purity levels that surpass existing technologies, ensuring sterility and therapeutic suitability without additional sterilization.
Implementation Method 1
an excitation light source to emit an excitation beam to generate fluorescence light from target particles
Implementation Method 2
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.