Optical Tomography Specimen Enrichment Using Multi-Stage FACS Gating
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Solution Overview
Problem
Existing optical computed tomography systems face inefficiencies in processing large samples of biological specimens, particularly sputum, due to the need to analyze millions of cells, many of which are non-diagnostic, leading to excessive resource usage and time consumption.
Innovation Solution
A method and system for enriching specimens using fluorescence-activated cell sorting (FACS) with controlled specimen treatment and multiple gate construction to isolate target cells, such as bronchial epithelial cells, by constructing and optimizing primary and secondary gates based on scatterplot data to achieve high enrichment efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If all objects in the sample are processed for 3D reconstruction, then complete analysis coverage is achieved, but processing time and resource consumption become excessive
Solution Approach 1:
The patent extracts and isolates only the target objects (bronchial epithelial cells) from the mixed specimen using FACS gating based on specific optical properties. By separating target cells from non-target cells (oral squamous cells, immune cells, debris) before 3D reconstruction, the system achieves complete analysis coverage of relevant cells while dramatically reducing processing time and resources by excluding irrelevant objects.
2Productivity
If sample enrichment is performed to eliminate non-target cells, then processing efficiency improves, but system complexity increases
Solution Approach 1:
The patent performs preliminary enrichment of target cells using FACS gating before the main 3D reconstruction process. By pre-sorting and isolating bronchial epithelial cells based on their unique optical scattering properties in a first scatterplot, the system prepares an enriched sample that streamlines subsequent analysis. This preliminary action increases processing efficiency while managing complexity through automated gating algorithms.
3Manufacturing precision
If multiple gating stages are implemented to achieve high enrichment efficiency, then target cell isolation improves, but processing complexity increases
Solution Approach 1:
The patent segments the gating process into distinct stages: first scatterplot gating to isolate target cells from non-target cells based on optical properties, and second scatterplot gating to further refine the enriched sample. This segmentation of the enrichment process into manageable steps achieves high target cell isolation efficiency (95% or greater purity) while controlling complexity through systematic, multi-stage filtering rather than a single complex gate.
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 significantly reduces processing time and resource consumption by enriching samples to exclude non-target cells, allowing for more efficient 3D reconstruction and analysis of target cells, thereby improving the sensitivity and specificity 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
generating a first scatterplot of the first set of 2D event data; locating a first set of the known target objects in the first scatterplot
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.