Cell Population Profiling for Rare Subset Detection at High Throughput
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Solution Overview
Problem
Current techniques for phenotypic analysis of cell populations, such as flow cytometry, mass cytometry, and single-cell sequencing, either provide low-resolution profiling of large numbers of cells with high throughput or high-resolution profiling of a small number of cells with low throughput, failing to effectively characterize rare cell subsets within heterogeneous populations.
Innovation Solution
A method involving sorting cells into multiple types based on marker expression, labeling with target-binding molecules, and analyzing using combinations of flow cytometry and mass cytometry, with the use of identifiers to enhance resolution and throughput.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If flow cytometry is used for high-throughput analysis, then the number of cells analyzed per second is high (25,000 cells/second), but the number of markers that can be analyzed is limited (～30 markers)
Solution Approach 1:
The patent combines flow cytometry and mass cytometry into a hybrid system that integrates the high throughput capability of flow cytometry with the high marker capacity of mass cytometry, enabling simultaneous analysis of thousands of cells with hundreds of markers
2Quantity of substance
If mass cytometry is used to increase the number of markers analyzed (～40 markers), then the throughput decreases to (500-1,000 cells/second)
Solution Approach 1:
The patent merges the advantages of both flow and mass cytometry by integrating flow cytometry's high cell throughput with mass cytometry's ability to detect numerous markers simultaneously, achieving both high marker capacity and high throughput
3Measurement precision
If single cell sequencing (CITE-Seq) is used for high-resolution profiling of hundreds of markers, then the throughput is much lower (a few thousand cells per experiment)
Solution Approach 1:
The patent integrates the high-resolution marker detection capability of sequencing-based methods with the high throughput of flow cytometry, enabling analysis of hundreds of markers across tens of thousands of cells simultaneously
4Measurement precision
If current techniques are used for phenotypic analysis, then either high throughput or high resolution can be achieved, but not both simultaneously
Solution Approach 1:
The patent creates a hybrid cytometry system that merges flow cytometry and mass cytometry technologies, enabling simultaneous achievement of high throughput and high resolution for comprehensive cell population analysis
Solution Approach 2:
The patent develops a multi-functional system that can perform both flow cytometry and mass cytometry analyses, providing universal capability to achieve high throughput or high resolution depending on the specific experimental requirements
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 high-throughput and high-resolution profiling of heterogeneous cell populations, allowing for the analysis of a greater number of molecules and cell types with improved sensitivity to rare cell populations.
Implementation Method 1
analysis by flow cytometry and mass cytometry. However, these techniques have limitations. Flow cytometry employs laser excitation and subsequent detection and evaluation of light scatter and emission spectra
Implementation Method 2
Cells can be stained with antibodies that bind specifically to molecules of interest, and the antibodies can be labelled e.g. by conjugation to a fluorophore. Cells expressing the molecules of interest can be identified by detection of signal emitted by the fluorophore following excitation of the cell with a laser within the excitation spectrum of the fluorophore
Implementation Method 3
Analysis of populations of cells by mass cytometry employs antibodies conjugated to elemental metal isotopes. Stained cells are introduced into an inductively coupled mass spectrometer by droplet nebulisation, and the elemental isotopes are detected and correlated with the presence of the molecules
Data Source
AI summary
Methods for evaluating the expression of one or more molecules of interest by cell types of interest within a heterogeneous population of cells are provided, comprising: (i) sorting cells of the heterogeneous population of cells into two or more cell types based on expression of one or more cell type markers by the cells; (ii) labelling the cells with target-binding molecules capable of binding to one or more molecules of interest; and (iii) analysing the cells in order to determine expression of the one or more molecules of interest by the one or more cell types of interest.