Abstract
The PAMONO-sensor (plasmon assisted microscopy of nano-objects) demonstrated an ability to detect and quantify individual viruses and virus-like particles. However, another group of biological vesicles—microvesicles (100–1000 nm)—also attracts growing interest as biomarkers of different pathologies and needs development of novel techniques for characterization. This work shows the applicability of a PAMONO-sensor for selective detection of microvesicles in aquatic samples. The sensor permits comparison of relative concentrations of microvesicles between samples. We also study a possibility of repeated use of a sensor chip after elution of the microvesicle capturing layer. Moreover, we improve the detection features of the PAMONO-sensor. The detection process utilizes novel machine learning techniques on the sensor image data to estimate particle size distributions of nano-particles in polydisperse samples. Altogether, our findings expand analytical features and the application field of the PAMONO-sensor. They can also serve for a maturation of diagnostic tools based on the PAMONO-sensor platform.
Highlights
IntroductionThe PAMONO-sensor (plasmon assisted microscopy of nano-objects) harnesses a surface plasmon resonance (SPR) phenomena for the imaging of biological and non-biological nano-objects [1,2]
The PAMONO-sensor harnesses a surface plasmon resonance (SPR) phenomena for the imaging of biological and non-biological nano-objects [1,2]
We demonstrate for the first time the ability of the PAMONO-sensor to perform a real-time observation of the binding of label-free individual microvesicles to the sensor surface functionalized by protein A/G and anti-target antibody
Summary
The PAMONO-sensor (plasmon assisted microscopy of nano-objects) harnesses a surface plasmon resonance (SPR) phenomena for the imaging of biological and non-biological nano-objects [1,2]. In the conventional SPR approach, the layers of biomolecules formed on a gold sensor surface alter the refractive conditions near the sensor surface. These alterations serve as a basis for the real-time measurements of the binding efficiency of biomolecules during determination of affinity constants or concentration measurements. Sensors 2017, 17, 244 long time, plasmonic sensors were not deemed as effective instruments for the quantification of the binding of individual nano-scale particles to the sensor surface. It is not surprising that SPR-based sensors were recently [10,11] tried out in studies focused on detection and quantification of another group of biological nano-vesicles—extracellular vesicles. Non-apoptotic EVs can carry different biologically active cargo and may be divided according to their size into two groups: exosomes (20–100 nm) and microvesicles (MVs) (100–1000 nm) [12,13]
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