Flow Cytometry Gate Calibration for Stable Particle Subpopulation Sorting
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Solution Overview
Problem
Current flow cytometry systems for defining subpopulations are subjective and variable, leading to inconsistencies in yield and purity, especially in sex sorting cytometers, and fail to adapt to operational changes due to factors like temperature or pressure.
Innovation Solution
A self-calibrating identification system that refines subpopulation definitions by integrating additional information from particle mixtures, continuously updating critical ranges of values, and using a control apparatus to assign subpopulation classifications based on measured characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If manual visual inspection and manual gate drawing are used to define subpopulations, then operator flexibility and visual judgment can be applied, but subjectivity and variability between operators and runs increase
Solution Approach 1:
The system performs self-calibration by automatically defining gates and subpopulations based on algorithmic analysis of particle characteristics. The control apparatus autonomously identifies critical ranges and updates gate definitions without requiring manual operator intervention, thereby eliminating subjectivity while maintaining operational simplicity.
Solution Approach 2:
The system dynamically adjusts gate parameters and critical ranges based on measured particle characteristics. By changing the parameters from fixed manual values to dynamically calculated values based on particle data distributions, the system achieves both automation and consistency while preserving the ability to adapt to different sample types.
2Device complexity
If static gate definitions are used for subpopulation identification, then simplicity and ease of implementation are maintained, but adaptability to operational changes (temperature, pressure) is lost
Solution Approach 1:
The gate definitions transition from static to dynamic through continuous self-calibration. The control apparatus periodically recalculates critical ranges and updates gate parameters based on current particle measurements, allowing the system to adapt to environmental changes while maintaining a simple implementation framework.
Solution Approach 2:
The system implements feedback loops where particle measurement data is continuously fed back to the control apparatus, which then adjusts gate definitions accordingly. This feedback mechanism enables automatic adaptation to operational changes without increasing overall system complexity.
3Ease of manufacture
If manual gate drawing is used to define subpopulations, then initial setup is simple, but yield and purity consistency decreases
Solution Approach 1:
The system performs self-calibration to automatically optimize gate definitions for maximum yield and purity. By eliminating manual gate drawing in favor of algorithmic determination based on particle characteristics, the system maintains ease of initial setup while dramatically improving consistency of sorting results.
4Stability of the object's composition
If fixed critical ranges are assigned to subpopulations, then stability and predictability are achieved, but measurement precision and accuracy decrease
Solution Approach 1:
Critical ranges transition from fixed to dynamically adjusted values through continuous self-calibration. The control apparatus periodically recalculates optimal critical ranges based on current particle data, achieving both stability through systematic methodology and precision through data-driven adjustments.
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
Enhances operational consistency, reduces variability, and improves yield and purity by dynamically adapting to changes in the cytometric environment.
Implementation Method 1
As the stained sperm cells (a plurality of particles) are irradiated, the fluorochrome in the plurality of particles emits a responsive fluorescent light
Implementation Method 2
The detection component may generate an electrical, analog signal in response to the received light
Data Source
AI summary
Systems and methods are provided for identification and discrimination of subpopulations within a mixture of particles. The systems and methods implement continuous calibration of the classification of particles within the mixture to provide consistency in operation and to reduce inter- and intra-batch processing variation. The systems and methods produce advantageously sorted particle products.