Multi-Wavelength Spectral Scatter Flow Cytometry for Nanoparticle Detection

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Solution Overview

Problem

Current methods in flow cytometry are inadequate for detecting and sorting single nanoparticles and extracellular vesicles due to limitations in available tools and reagents, particularly for particles smaller than 500 nm, which are often undetectable due to signal loss in debris and electronic noise.

Innovation Solution

A microfluidic apparatus utilizing a multi-wavelength illumination source and detector system that elastically side-scatters light to determine the presence of nanotags, including single nanotags, by comparing side-scatter intensity characteristics with predetermined profiles, allowing for the detection and sorting of nanoscale particles and extracellular vesicles.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional flow cytometry methods are used, then detection of larger particles is possible, but detection of nanoparticles smaller than 500 nm is lost in signal debris and electronic noise

Engineering Contradiction:
Improvedetection sensitivityVSAvoidsignal loss in debris and electronic noise
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The detection system segments the broad optical spectrum into multiple wavelength channels, with each channel optimized for detecting specific nanoparticle sizes or types. This spectral segmentation allows the system to resolve nanoparticle signals from background noise by analyzing scattering patterns at different wavelengths simultaneously.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The system changes the illumination wavelength parameter across multiple channels to optimize detection for different nanoparticle types. By varying the wavelength and analyzing the resulting elastic scattering patterns, the system can distinguish nanoparticle signals from debris and electronic noise through spectral deconvolution algorithms.

Inventive Principle:
Principle #35Parameter changes

2Measurement precision

If multi-wavelength illumination is used to detect nanotags, then detection precision improves, but device complexity increases

Engineering Contradiction:
Improvenanotag detection precisionVSAvoidmulti-wavelength illumination system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The multi-wavelength illumination system serves multiple functions: it enables detection of different nanoparticle types, provides spectral information for identification, and allows multiplexed detection of multiple nanotag types simultaneously. This multi-functionality justifies the increased complexity by delivering comprehensive nanoparticle characterization capabilities.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent introduces computational algorithms as an intermediary that processes the multi-wavelength scattering data. These algorithms deconvolute the complex spectral signals and identify nanoparticle presence and type, effectively managing the complexity of the multi-wavelength system through software-based signal interpretation.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Measurement precision

If elastic side-scatter detection is used, then single nanoparticle detection is enabled, but detection of multiple nanotag types simultaneously becomes challenging

Engineering Contradiction:
Improvesingle nanoparticle detection capabilityVSAvoidmultiplexed nanotag type detection
Core Design Contradiction:
Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The system adds the wavelength dimension to elastic scattering detection, transforming it from a single-parameter measurement into a multi-dimensional spectral analysis. By detecting scattering intensity across multiple wavelengths simultaneously, the system can distinguish between different nanotag types based on their unique spectral scattering patterns while maintaining single nanoparticle detection sensitivity.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

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 the detection and sorting of single nanoparticles and extracellular vesicles with diameters as small as 100 nm or smaller, improving the ability to characterize and analyze nanoscale particles based on specific attributes, enhancing the sensitivity and accuracy of flow cytometry.

Implementation Method 1

the multi-wavelength detection beam comprises light that is elastically side-scattered by an interaction between the multi-wavelength illumination beam and the nanotags in the microfluidic target

Methodology Applied
Scientific EffectElastic side-scattering: Scattering

Data Source

PatentEP3701246B1Optical configuration methods for spectral scatter flow cytometry
Publication Date: 2021.02.24 THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY DEPARTMENT OF HEALTH & HUMAN SERVICES
  • EP3701246B1 patent drawingFigure 1
  • EP3701246B1 patent drawingFigure 2A~2D
  • EP3701246B1 patent drawingFigure 3A~3C

AI summary

Apparatus include an illumination source configured to produce and direct a multi-wavelength illumination beam to a microfluidic target that can include nanotags, a detector configured to receive a multi-wavelength detection beam from the microfluidic target and to produce a detection signal, wherein the multi-wavelength detection beam comprises light that is elastically side-scattered by an interaction between the multi-wavelength illumination beam and the nanotags in the microfluidic target, and a processor configured to receive the detection signal and to determine the presence of the nanotags in the microfluidic target by comparing multiple wavelength side-scatter intensity characteristics of the detection signal with predetermined multi-wavelength elastic side-scatter intensity profiles of one or more nanotag types. Methods are also disclosed that determine the presence of different nanotags responsive to a multi-wavelength detection beam based on a detected signal and predetermined multi-wavelength elastic side-scatter intensity profiles for different nanotag types.