Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles

• Autor(a) principal: Calado, Madalena Rosa Correia
• Data de Publicação: 2022
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10362/139625
• topic: microfluidics; extracellular vesicles; pressure-driven flow; nanoparticles; resistive-pulse sensing; Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia

Extracellular Vesicles (EVs) are highly involved in several biological processes within the human body. However, their involvement is not exclusive to conventional bodily functions since these vesicles can promote the spread and development of diseases due to their role in cell-to-cell communications. Thus, EVs have be-come serious biomarker candidates for the detection of pathological agents. Despite this, the current methods used to isolate and characterize EVs are often laborious and involve bulky laboratory instruments, hindering their routine use in healthcare. Therefore, developing new techniques to characterize them properly is of great interest. To tackle this problem, we studied the combination of Resistive-Pulse Sensing with pressure-driven flow in two different nanopores – one formed from glass capillaries and another integrated into microfluidic chips. First, we characterized the transport using model particles, which generated current spikes when moving through the pore. These experiments were followed by using biological samples, such as human serum and purified EVs. The results show that particle size strongly influences the current spike amplitude and width. Furthermore, it is possible to link relative current shift with the expected particle diameter, while frequency relates to particle concentration. By analyzing the relative current shift, we identified different population sizes in the complex biological samples. This work extends the knowledge on particle detection and sizing techniques but is also helpful to under-stand the possibilities of using Resistive Pulse Sensing in the analysis of EVs.