Spectral Flow Cytometry Panel for AML Detection
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Solution Overview
Problem
Current flow cytometry technologies face limitations in performing high-throughput, in-depth analysis of immune cell populations, particularly in cases of limited sample availability, such as in clinical trials, where detailed characterization of immune responses and identification of biomarkers are needed for cancer, autoimmunity, and infectious diseases.
Innovation Solution
A full spectrum flow cytometry system with multiple lasers and detectors is developed, enabling the measurement of entire fluorochrome emission spectra, allowing for the combination of multiple fluorescently labeled antibodies and the use of a similarity and complexity index to select compatible fluorochromes, which enhances the ability to distinguish between fluorophores and perform multiparametric analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional flow cytometry is used, then the system is simple to operate, but the measurement precision and depth of immune cell analysis is insufficient
Solution Approach 1:
The patent transitions from conventional flow cytometry to mass cytometry by adding the dimension of time-resolved detection. The system measures fluorescence emission at multiple time points (immediate, delayed, and sustained emission) to distinguish between different fluorochrome types, enabling deeper immune cell analysis without increasing spatial or structural complexity of the instrument
2Adaptability or versatility
If multiple fluorescently labeled antibodies are combined, then the ability to distinguish fluorophores and perform multiparametric analysis is enhanced, but the device complexity increases
Solution Approach 1:
The patent introduces time-resolved detection as an intermediary mechanism to resolve fluorophore overlap. By measuring emission at different time points, the system acts as a mediator that separates signals from different fluorochromes based on their temporal emission characteristics, enabling multiparametric analysis without requiring complex spectral unmixing hardware
Solution Approach 2:
The system changes the detection parameter from static fluorescence intensity measurement to dynamic time-resolved fluorescence measurement. This parameter change allows the system to distinguish between fluorophores with different emission kinetics, enhancing multiparametric analysis capability while using standard flow cytometry hardware
3Quantity of substance
If sample volume is limited, then the loss of substance is minimized, but the productivity and throughput of analysis is reduced
Solution Approach 1:
The patent implements continuous analysis by processing cells in suspension through the flow cytometer continuously. The automated flow-based system maintains continuous detection without requiring manual sample manipulation or preparation steps, maximizing throughput from limited samples by eliminating idle time and enabling analysis of every cell in the limited sample volume
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 approach enables the detection of up to 40 colors in a single sample, providing detailed characterization of immune cell populations and overcoming the limitations of sample availability, with improved resolution and accuracy in immune cell analysis.
Implementation Method 1
Flow cytometers utilize lasers as light sources to produce both scattered and fluorescent light signals that are read by detectors such as photodiodes or photomultiplier tubes
Implementation Method 2
Flow cytometers utilize lasers as light sources to produce both scattered and fluorescent light signals
Implementation Method 3
In hydrodynamic focusing, the sample fluid is enclosed by an outer sheath fluid and injected through a nozzle or cuvette
Implementation Method 4
Under optimal conditions (laminar flow) there is no mixing of the central fluid stream and the sheath fluid
Data Source
AI summary
In one embodiment, a method of building an optimized color flow high parameter reagent and reagent kit cytometry panel for detection of aberrant cells in acute myeloid leukemia is disclosed using a spectral flow cytometer with a least three excitation lasers and thirty-eight color detectors. In another embodiment, a graphical user interface is disclosed generated by a server computer from a fluorochrome database and displayed by a client computer to assist in the selection of a set of fluorochromes for use in an assay to analyze biological samples. The GUI can display spectra graphs to visually show how fluorochromes may overlap and can generate similarity indexes for the paired fluorochrome interference and a complexity index for overall many to many interferences generated by a selected group or set of fluorochromes.