Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets

Geetika Raizada; Balasubramaniam Namasivayam; Sameh Obeid; Benjamin Brunel; Wilfrid Boireau; Eric Lesniewska; Celine Elie-Caille

doi:10.3791/64210

Plataforma analítica multimodal en un chip de resonancia de plasmón de superficie multiplexada pa...

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Multimodal Analytical Platform on a Multiplexed Surface Plasmon Resonance Imaging Chip for the Analysis of Extracellular Vesicle Subsets

Plataforma analítica multimodal en un chip de resonancia de plasmón de superficie multiplexada para el análisis de subconjuntos de vesículas extracelulares

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06:12 min

March 17, 2023

DOI: 10.3791/64210-v

Geetika Raizada¹, Balasubramaniam Namasivayam², Sameh Obeid³, Benjamin Brunel¹, Wilfrid Boireau¹, Eric Lesniewska⁴, Celine Elie-Caille¹

¹Université de Franche-Comté, CNRS UMR-6174,FEMTO-ST Institute, ²Lille Neuroscience & Cognition research centre, ³Institut Galien Paris-Saclay, CNRS UMR-8612 , Université Paris-Saclay, ⁴Interdisciplinary Lab Carnot Bourgogne LICB, CNRS UMR-6303,Université de Bourgogne Franche-Comté

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Overview

This study presents a novel multiparametric analytical platform for characterizing extracellular vesicle (EV) subsets with high throughput. The platform combines multiplexed biosensing methods with atomic force microscopy and Raman spectroscopy to analyze EVs in real-time, focusing on their phenotypes, size profiles, and nanomechanical properties.

Key Study Components

Research Area

Extracellular vesicle characterization
Multiparametric analytical techniques
Biosensing methods

Background

Complexity of EVs makes subpopulation identification challenging.
Importance of EVs as biomarkers for pathologies and drug delivery.
Need for sensitive detection methods in various biological samples.

Methods Used

Combines multiplexed biosensing, atomic force microscopy, and Raman spectroscopy.
Utilizes biofunctionalized microarrays for detecting EVs in real-time.
Label-free approach to minimize interference with EV characterization.

Main Results

The technique shows effective detection of EVs based on size and composition.
High-resolution imaging reveals the diameter of EVs ranging from 30 to 300 nanometers, predominantly around 60 nanometers.
Raman spectra indicate lipid membrane composition, confirming the method's effectiveness.

Conclusions

The study successfully demonstrates a reliable method for extensive EV characterization.
This work has significant implications for biomarker research and therapeutic applications.

Frequently Asked Questions

What are extracellular vesicles?

Extracellular vesicles (EVs) are small membrane-bound particles secreted by cells that play roles in intercellular communication and can serve as biomarkers.

How does the described platform improve EV detection?

The platform improves detection by combining multiple analytical techniques for enhanced sensitivity and specificity in identifying EV subsets.

Why is a label-free method important for this analysis?

A label-free method allows real-time monitoring of EV interactions without the potential interference caused by labeling agents.

What types of samples can this method analyze?

This method can analyze various biological samples, including conditioned media and blood plasma.

What are the potential applications of this research?

The findings can be applied in diagnostics as biomarkers for diseases and in drug delivery systems utilizing EVs.

How does the biochip preparation affect the outcome?

Careful biochip preparation is crucial for ensuring reproducible and accurate detection of EVs during analysis.

Can traditional methods be used alongside this new technique?

Yes, traditional methods like Western blotting and nanoparticle tracking can be used for correlation and validation of findings.

Este artículo propone una nueva generación de plataformas analíticas multiparamétricas con mayor rendimiento para la caracterización de subconjuntos de vesículas extracelulares. El método se basa en una combinación de métodos de biodetección multiplexada con análisis metrológicos y morfomecánicos por microscopía de fuerza atómica, junto con espectroscopia Raman, para calificar objetivos vesiculares atrapados en un biochip de microarray.

La naturaleza compleja de los EV hace que sea difícil reconocer la subpoblación de interés, y este protocolo puede detectarlos con su fenotipo, perfil de tamaño y propiedades nanomecánicas. Esta combinación de técnica es un método sin etiqueta, que nos permite detectar EV en tiempo real desde los medios de afección y el plasma sanguíneo. Esta técnica permite caracterizar profundamente el subconjunto de nanopartículas de interés, con el fin de ser utilizadas como bioindicadores de patología, o también utilizadas como nanopartículas para la administración de fármacos.

Dado que es un método muy sensible, es imperativo prestar atención a la preparación del biochip antes de inyectar su muestra de interés. Después de recubrir y funcionalizar los chips, incubar el biochip en una mezcla de 200 milimolares de etil dimetilaminopropilcarbodiimida/N-hidroxisuccinimida y 50 milimolares por litro de N-hidroxisuccinimida durante al menos 30 minutos en la oscuridad a temperatura ambiente. Usando el observador, agregue 300 nanolitros de solución de ligando e incube el biochip bajo un baño sónico durante 30 minutos.

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