Plasmonic Scattering Microscopy Exosome Analysis
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Solution Overview
Problem
Current SPR microscopy techniques face limitations such as low spatial resolution due to a parabolic tail-shaped point spread function, limited throughput due to an expensive total internal reflection fluorescence objective with a semi-circular field of view, and difficulty in multiplexed protein marker analysis on a small field of view.
Innovation Solution
The implementation of plasmonic scattering microscopy (PSM) technology, which provides a Gaussian distributed point spread function for high spatial resolution, automatic image processing with conventional software, and a significantly larger field of view compared to SPR microscopy and nanoparticle tracking analysis instruments, enabling high throughput analysis of exosomes and multiplexed protein marker analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If SPR microscopy is used for exosome analysis, then biomarker detection capability is provided, but spatial resolution deteriorates due to parabolic tail-shaped point spread function
Solution Approach 1:
The system separates the detection functions into two independent modules: SPR microscopy for biomarker detection and NTA for size distribution analysis. This segmentation allows each module to optimize for its specific function without compromising the other, resolving the spatial resolution limitation of SPR microscopy while maintaining biomarker detection capability.
2Measurement precision
If total internal reflection fluorescence objective is used in SPR microscopy, then biomarker analysis capability is enabled, but throughput deteriorates due to limited field of view
Solution Approach 1:
The system transitions from a single-dimension approach (SPR microscopy alone) to a multi-dimensional integrated platform that combines SPR microscopy with NTA. This dimensional expansion allows simultaneous acquisition of biomarker data and size distribution data across a larger field of view, thereby increasing throughput while maintaining analytical capability.
3Measurement precision
If SPR microscopy with small field of view is used, then biomarker detection is achieved, but multiplexed protein marker analysis becomes difficult
Solution Approach 1:
The integrated platform provides universal functionality by combining SPR microscopy for biomarker detection with NTA for size distribution analysis. This multi-functional system can perform both single-marker and multiplexed protein marker analysis, as well as size distribution measurements, making it adaptable to various analytical requirements without being constrained by the limited field of view of SPR microscopy alone.
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
PSM achieves high spatial resolution and throughput for exosome analysis, allowing for efficient multiplexed protein marker analysis and monitoring of antibody binding kinetics without the need for labels, thus overcoming the limitations of existing SPR microscopy techniques.
Implementation Method 1
detecting light scattered by the surface-bound exosomes to produce a set of exosome imaging data
Implementation Method 2
PSM was first developed to realize label-free single-molecule imaging on SPR microscopy
Data Source
AI summary
Provided herein are methods of detecting exosomes, including unlabeled exosomes. In some embodiments, the methods include disposing a fluidic sample that comprises a plurality of exosomes in a chamber that is positioned at least partially within a fluidic device in which an inner surface of the chamber comprises a first set of exosome binding moieties that are capable of binding the exosomes. In some embodiments, the methods also include binding a portion of the plurality of exosomes to a portion of the first set of exosome binding moieties to produce surface-bound exosomes, introducing an incident light toward the inner surface of the chamber prior to, concurrent with, and/or after, producing the surface-bound exosomes, and detecting light scattered by the surface-bound exosomes to produce a set of exosome imaging data. Related fluidic devices, systems, and computer readable media are also provided.