FACS Gating for Enriched Optical Tomography Cell Analysis
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Solution Overview
Problem
Existing optical computed tomography systems face inefficiencies in processing large volumes of specimen samples for lung cancer detection, as they process all objects, including non-target cells, leading to excessive resource consumption and time, necessitating a method to enrich samples and eliminate extraneous objects.
Innovation Solution
A method and system for enriching specimens using fluorescence-activated cell sorting (FACS) by constructing and optimizing gates based on scatterplots of 2D event data to isolate target cells, ensuring predetermined percentages of target objects are captured within defined gates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If all objects in the specimen are processed for optical tomography analysis, then comprehensive detection coverage is achieved, but processing time and resource consumption increase excessively
Solution Approach 1:
The patent applies the extraction principle by removing non-target cells and extraneous objects from the specimen before optical tomography analysis. The system identifies and extracts only the relevant target cells (such as lung cancer cells) from the mixed cell population, eliminating the need to process all objects. This significantly reduces processing time and computational resources while maintaining detection coverage for the clinically relevant cell types.
Solution Approach 2:
The patent applies segmentation by dividing the specimen processing into distinct stages: initial specimen preparation, FACS-based cell sorting to separate target cells from non-target cells, and subsequent optical tomography analysis only on the sorted target cell population. This segmented approach allows the system to focus computational resources on analyzing only the relevant cell subsets rather than processing the entire heterogeneous specimen uniformly.
2Reliability
If all objects in the specimen are processed for optical tomography analysis, then comprehensive detection coverage is achieved, but resource consumption increases excessively
Solution Approach 1:
The system extracts and isolates only the target cell population from the mixed specimen using FACS sorting based on fluorescently labeled antibodies. By removing non-target cells (such as oral squamous cells and immune cells) before optical tomography analysis, the system dramatically reduces the number of objects requiring computationally intensive 3D reconstruction and analysis, thereby reducing energy and processing resource consumption while maintaining comprehensive detection coverage for the clinically relevant target cells.
3Reliability
If FACS gating is optimized to capture all target cells, then capture rate increases, but gate complexity and optimization time increase
Solution Approach 1:
The patent applies preliminary action by using fluorescently labeled antibodies that bind to specific cell surface markers on target cells before FACS analysis. This pre-labeling step creates distinct fluorescent signals that simplify gate construction in the FACS instrument, allowing operators to set gates based on fluorescence intensity thresholds rather than having to optimize complex multi-parameter gates. The preliminary staining action reduces the complexity of subsequent gating operations while maintaining high capture rates for the target cell population.
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 method enhances the efficiency of optical tomography analysis by enriching samples, reducing processing time and resources, while maintaining high capture rates of target cells, thereby improving the accuracy and speed of lung cancer detection.
Implementation Method 1
introducing a controlled specimen including a plurality of known objects treated with an Ab cocktail into a fluorescence-activated cell sorter (FACS)
Implementation Method 2
optical computed tomography system begins with specimen collection and preparation
Implementation Method 3
Images of objects, such as cells, in the specimen are collected while the cells are rotated around 360-degrees relative to the image collection optics
Data Source
AI summary
A method for enhancing gating performance of a cell sorter to prepare an enriched specimen for optical tomography cell analysis includes introducing a specimen into a FACS to generate 2D event data; generating a first scatterplot of the 2D data; identifying target objects; constructing a boundary within the first scatterplot to produce a first gate; counting target objects within the first gate; comparing the number of target objects within the first gate to a first predetermined value and adjusting the first gate as necessary. A boundary around a set of target objects is constructed in a second scatterplot to produce a subset second gate and target objects within the second gate are counted and the count compared to a second predetermined value. When a boundary around target objects meets specifications the first and second gates are stored in memory and used to enrich patient specimens.