Cell Detection via Flow Cytometry and Imaging Verification
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Solution Overview
Problem
The flow cytometry method often results in nonspecific sorting of cells due to staining nonspecificity, leading to inaccurate gene analysis, and sometimes multiple cells being sorted into a single well, which affects the effectiveness and accuracy of DNA amplification and subsequent gene analysis.
Innovation Solution
A cell detection method that involves sorting target cells into a container with multiple wells using flow cytometry, followed by imaging to obtain additional information from the cells, allowing for precise determination and isolation of target cells for accurate gene analysis, which includes staining cells before sorting, correlating well positions with sorting and imaging data, and using centrifugation to move cells to the well bottom for imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If flow cytometry method is used to sort target cells, then cell sorting efficiency is improved, but sorting accuracy deteriorates due to nonspecific staining
Solution Approach 1:
The patent introduces an intermediary imaging verification step between flow cytometry sorting and subsequent analysis. The imaging device captures images of sorted cells to verify sorting accuracy, acting as a mediator that detects and corrects sorting errors caused by nonspecific staining without reducing the efficiency of the flow cytometry method itself.
Solution Approach 2:
The system implements feedback by using imaging information to verify and correct sorting results. The imaging device provides feedback on whether cells were correctly sorted into wells, allowing for identification and correction of missorted cells, thereby improving sorting accuracy while maintaining the high throughput of flow cytometry.
2Speed
If flow cytometry sorting is performed based on fluorescence intensity, then sorting speed is improved, but reliability of gene analysis deteriorates due to missorting
Solution Approach 1:
The imaging device serves as an intermediary verification step that checks the reliability of flow cytometry sorting without reducing sorting speed. By capturing images of sorted cells and verifying their correct placement in wells, the system ensures reliable gene analysis results while maintaining high sorting throughput.
Solution Approach 2:
The patent replaces reliance solely on flow cytometry's mechanical sorting mechanism with an additional optical verification system. The imaging device provides a second layer of verification using optical methods to confirm correct sorting, thereby enhancing reliability without sacrificing the speed advantage of flow cytometry.
3Productivity
If multiple cells are sorted into one well, then throughput is improved, but measurement accuracy deteriorates
Solution Approach 1:
The imaging system provides feedback on the number of cells in each well, allowing the system to identify wells containing multiple cells. This feedback mechanism enables correction or re-sampling of multi-cell wells, ensuring measurement accuracy is maintained while preserving the high throughput capability of sorting multiple cells into arrays of wells.
Solution Approach 2:
The patent replaces reliance on flow cytometry's mechanical assumption of one-cell-per-well with an optical verification system. The imaging device directly observes and counts cells in each well, providing accurate measurement information that corrects for instances where multiple cells were accidentally sorted into a single well, thereby maintaining both throughput and accuracy.
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
This method enables effective and accurate gene analysis by ensuring only target cells are analyzed, improving the accuracy of DNA amplification and reducing contamination, thereby enhancing the reliability of genetic information obtained.
Implementation Method 1
the information such as forward-scattered light, side-scattered light, and fluorescence intensity is obtained from cells
Implementation Method 2
the information such as forward-scattered light, side-scattered light, and fluorescence intensity is obtained from cells
Implementation Method 3
the imaging step includes a step of moving the cells to a bottom surface of each of the wells by centrifugation
Data Source
AI summary
Provided is a cell detection method which makes it possible to effectively and accurately perform gene analysis only on target cells. The cell detection method includes a sorting step of obtaining first information derived from cells in a sample solution by using a flow cytometry method and sorting target cells into a container having arrays of wells each having an opening based on the first information, an imaging step of imaging the cells sorted into the container, and a determination step of obtaining second information derived from cells based on the image of the cells captured by the imaging step and determining cells to be analyzed from the sorted cells.