Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles
Autor(a) principal: | |
---|---|
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 |
Resumo: | 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. |
id |
RCAP_d3a98c73302500aa5884caebc4fc4ee7 |
---|---|
oai_identifier_str |
oai:run.unl.pt:10362/139625 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesiclesmicrofluidicsextracellular vesiclespressure-driven flownanoparticlesresistive-pulse sensingDomínio/Área Científica::Engenharia e Tecnologia::NanotecnologiaExtracellular 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.As Vesiculas Extracelulares (EVs) estão envolvidas em diversos processos biológicos do corpo humano. No entanto, devido ao papel destas vesiculas na comunicação entre células, este envolvimento não é exclusivo às funções corporais convencionais, uma vez que podem promover a disseminação de doenças. Assim, o inte-resse para usar as EVs como biomarcadores de agentes patológicos tem vindo a crescer. Apesar disto, os mé-todos atualmente implementados para isolar e caracterizar EVs são, muitas vezes, trabalhosos e envolvem equipamentos de laboratório volumosos, dificultando a sua implementação no típico contexto hospitalar. As-sim sendo, existe um grande interesse em desenvolver novas técnicas capazes de as caracterizar adequada-mente. Para combater este problema, estudámos a combinação de Deteção de Pulso Resistivo com fluxo gerado por pressão em dois nanoporos diferentes – um formado a partir de capilares de vidro e outro integrado em dispositivos de microfluídica. Primeiro, caracterizámos o transporte usando partículas modelo, que ao atraves-sarem o poro geram picos de corrente. Estes testes foram seguidos pela análise de amostras biológicas, como sérum humano e EVs purificadas. Os resultados mostram que o tamanho da partícula influencia fortemente a amplitude e a largura do pico de corrente. Além disso, é possível relacionar a alteração relativa da corrente com o diâmetro esperado da partícula, enquanto a frequência está relacionada com a concentração das partículas. Ao analisar esta alteração relativa da corrente conseguimos identificar nas amostras biológicas a existência de populações com diferentes dimensões. O presente projeto expande o conhecimento sobre técnicas de deteção e dimensionamento de partículas, sendo também útil para compreender as diferentes possibilidades de usar Deteção de Pulso Resistivo na análise de EVs.Ainla, AlarÁguas, HugoRUNCalado, Madalena Rosa Correia2022-06-08T12:56:47Z2022-022022-02-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/139625enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:16:52Zoai:run.unl.pt:10362/139625Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:49:27.927182Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
title |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
spellingShingle |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles Calado, Madalena Rosa Correia microfluidics extracellular vesicles pressure-driven flow nanoparticles resistive-pulse sensing Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
title_short |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
title_full |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
title_fullStr |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
title_full_unstemmed |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
title_sort |
Pressure-Driven Resistive Pulse Nanopore Sensing for the Characterization of Extracellular Vesicles |
author |
Calado, Madalena Rosa Correia |
author_facet |
Calado, Madalena Rosa Correia |
author_role |
author |
dc.contributor.none.fl_str_mv |
Ainla, Alar Águas, Hugo RUN |
dc.contributor.author.fl_str_mv |
Calado, Madalena Rosa Correia |
dc.subject.por.fl_str_mv |
microfluidics extracellular vesicles pressure-driven flow nanoparticles resistive-pulse sensing Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
topic |
microfluidics extracellular vesicles pressure-driven flow nanoparticles resistive-pulse sensing Domínio/Área Científica::Engenharia e Tecnologia::Nanotecnologia |
description |
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. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-06-08T12:56:47Z 2022-02 2022-02-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/139625 |
url |
http://hdl.handle.net/10362/139625 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799138093457997824 |