Flow Cytometer Waveform Characterization Using Basis Functions
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Solution Overview
Problem
Current flow cytometry techniques fail to effectively utilize the vast amount of data obtained for each sample, often discarding detailed waveform information, which can lead to missed identification of cell doublets, debris, and valuable cell shape data.
Innovation Solution
The method involves generating a waveform from detected radiated light and transforming it using basis functions like wavelet or Fourier transformations to obtain coefficients that characterize particle properties, enabling detailed analysis of physical and biological properties without requiring prior knowledge or extensive raw data storage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If detailed waveform information is retained for analysis, then measurement precision of particle properties is improved, but loss of time in processing and storing data increases
Solution Approach 1:
The patent extracts only the essential characteristics of waveforms by transforming them into coefficients (e.g., through Fourier or wavelet transforms). Instead of retaining and processing entire detailed waveforms, the system extracts key coefficient values that capture the essential particle properties, thereby reducing data volume and processing time while maintaining measurement precision.
Solution Approach 2:
The patent transforms waveform data from the time domain to the frequency domain or wavelet domain, changing the parameter representation. By converting continuous waveform signals into discrete coefficient parameters, the system reduces the dimensional complexity of the data while preserving the information needed for accurate particle characterization.
2Device complexity
If traditional threshold-based analysis is used, then device complexity is reduced, but loss of information about cell shape and doublets increases
Solution Approach 1:
The patent replaces the mechanical/threshold-based analysis approach with a mathematical transformation approach. Instead of using simple voltage thresholds to gate particles, the system applies Fourier or wavelet transforms to extract coefficient patterns that encode waveform shape information, thereby preserving details about cell shape and doublets while maintaining analytical simplicity.
3Reliability
If waveform smoothing algorithms are applied, then reliability of peak detection is improved, but loss of information on detailed waveform shapes increases
Solution Approach 1:
Rather than smoothing the entire waveform which would lose shape details, the patent extracts specific coefficient information from the transform domain that captures the essential shape characteristics needed for reliable peak detection and particle identification, preserving waveform information while achieving reliable detection.
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 allows for accurate characterization of particles by extracting detailed waveform characteristics, improving the identification of cell types and properties, and enabling real-time sorting without the need for fluorescent markers, thus enhancing data processing efficiency and accuracy.
Implementation Method 1
The optical signals, derived from radiated light, for example from emission of fluorescence or from light scatter, are converted into voltage-versus-time pulse waveforms through the operation of a detector, such as photodiode or photomultiplier detectors.
Data Source
AI summary
The invention relates to a method and system for characterizing particles using a flow cytometer comprising generating a waveform, as a digital representation of detected radiated light, and transforming said waveform using one or more basis functions and obtaining one or more coefficients characterizing the waveform. The one or more coefficients characterizing the waveform preferably correspond to particular properties of the particle(s), thereby enabling analysis of physical properties of the particles (such as size or shape) or biological properties of the particles, such as cell type, localization and/or distribution of molecules within the cell and/or on the cell surface, structural elements of the cell such as the nucleus or the cytoskeleton, antibody or antibody-fragment binding to the cell or cell morphology. Preferred embodiments of the invention relate to methods and systems in which the waveform is transformed by a wavelet transformation or Fourier transformation.