Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/23795 |
Resumo: | Fluorescence Lifetime Image Microscopy (FLIM) of nanostructured graphene substrates was used to measure the recently observed nanoscale distance-scaling of the fluorescence lifetime of dyes located in the vicinity of graphene sheets, at distances up to about 30 nm. The results were compared with a Resonant Energy Transfer (RET) theoretical model and used to establish an experimental fluorescence lifetime-to-nanoscale distance conversion function. In the following, this nano-optical relation was used for the design of a Deoxyribonucleic Acid (DNA) biosensor. Graphene was functionalized with fluorescently labeled DNA molecular beacons that unfold during hybridization with complementary DNA, and thereby change the distance of the fluorescent dye from the graphene surface. The spatial distribution of DNA molecular beacons binding to the surface of a graphene flake was studied, as well as the temporal kinetics of the hybridization reaction using time-lapse FLIM measurements. The results showed a vertical ascent of a fluorescent label relative to the graphene surface with a distance extension that is in accordance with the expected molecular length of the specific DNA sequence used. Interestingly, an intermediate state associated to a distance of a few nanometers was identified with a lifespan of about 85 minutes. The developed graphene-based DNA sensor was shown to enable optical detection of nanoscale distances in liquid media. These findings indicate that the fluorescence lifetime-based detection coupled with nanoscale interaction effects may find applications in various biosensing applications such as health and food-quality tracing. For the processing of FLIM data, several fluorescence lifetime calculation algorithms were compared and integrated into a specially designed and implemented analysis software toolbox in MATLAB. |
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Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime CharacterizationGraphenenanoscale distance sensingfluorescence lifetimeFLIMenergy transferDNA biosensingMATLABGrafenomedição de distância à nanoescalatempo de vida de fluorescênciaFLIMtransferência de energiabiosensor de ADNMATLABDomínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e TecnologiasFluorescence Lifetime Image Microscopy (FLIM) of nanostructured graphene substrates was used to measure the recently observed nanoscale distance-scaling of the fluorescence lifetime of dyes located in the vicinity of graphene sheets, at distances up to about 30 nm. The results were compared with a Resonant Energy Transfer (RET) theoretical model and used to establish an experimental fluorescence lifetime-to-nanoscale distance conversion function. In the following, this nano-optical relation was used for the design of a Deoxyribonucleic Acid (DNA) biosensor. Graphene was functionalized with fluorescently labeled DNA molecular beacons that unfold during hybridization with complementary DNA, and thereby change the distance of the fluorescent dye from the graphene surface. The spatial distribution of DNA molecular beacons binding to the surface of a graphene flake was studied, as well as the temporal kinetics of the hybridization reaction using time-lapse FLIM measurements. The results showed a vertical ascent of a fluorescent label relative to the graphene surface with a distance extension that is in accordance with the expected molecular length of the specific DNA sequence used. Interestingly, an intermediate state associated to a distance of a few nanometers was identified with a lifespan of about 85 minutes. The developed graphene-based DNA sensor was shown to enable optical detection of nanoscale distances in liquid media. These findings indicate that the fluorescence lifetime-based detection coupled with nanoscale interaction effects may find applications in various biosensing applications such as health and food-quality tracing. For the processing of FLIM data, several fluorescence lifetime calculation algorithms were compared and integrated into a specially designed and implemented analysis software toolbox in MATLAB.Imagens de Microscopia de Tempo de Vida de Fluorescência (FLIM) de substratos de grafeno nano-estruturados foram usadas para a medição da recentemente observada alteração do tempo de vida de fluorescência de corantes quando localizados na vizinhança de grafeno para distâncias até cerca de 30 nm. Os resultados foram comparados com um modelo teórico de Transferência Ressonante de Energia (RET) e utilizados para o estabelecimento de uma função de calibração experimental entre tempo de vida de fluorescência e distâncias à nano-escala. Posteriormente, esta relação nano-ótica foi utilizada para o desenvolvimento de um biossensor de Ácido Desoxirribonucleico (ADN). Substratos de grafeno foram funcionalizados com sinais moleculares (molecular beacons) de ADN que se desdobram durante a hibridização com ADN complementar, alterando a distância do corante fluorescente à superfície de grafeno. A distribuição espacial da ligação de sinais moleculares de ADN à superfície do grafeno foi estudada, bem como a cinética de reação de hibridização, usando medições de FLIM por lapso de tempo. Os resultados mostraram a ascensão vertical dos marcadores fluorescentes relativamente à superfície, com uma deslocamento total que está de acordo com o comprimento molecular esperado para a sequência de ADN utilizada. Curiosamente, um estado intermedio associado a uma distância de alguns nanómetros foi identificado, tendo este uma duração de cerca de 85 minutos. Foi então demonstrado que o sensor de ADN desenvolvido permite a deteção ótica de distâncias à nano-escala em meio líquido. Estes resultados indicaram que a deteção baseada em tempo de vida de fluorescência, acoplada aos efeitos desta interação ótica à nano-escala pode ser utilizada em várias aplicações de biodeteção, tal como na saúde e no rastreamento da qualidade alimentar. Para o tratamento de dados de FLIM, vários algoritmos de cálculo de tempo de vida foram comparados e integrados num programa de análise especificamente desenhado e implementado para o efeito, em ambiente MATLAB.International Iberian Nanotechnology Laboratory – INLSilva, Ana G.Nieder, Jana B.RUNAdão, Ricardo Miguel Ribeiro2018-10-02T00:30:13Z2017-09-272017-09-27T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/23795enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:12:08Zoai:run.unl.pt:10362/23795Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:27:53.913717Repositório 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 |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
title |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
spellingShingle |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization Adão, Ricardo Miguel Ribeiro Graphene nanoscale distance sensing fluorescence lifetime FLIM energy transfer DNA biosensing MATLAB Grafeno medição de distância à nanoescala tempo de vida de fluorescência FLIM transferência de energia biosensor de ADN MATLAB Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
title_short |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
title_full |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
title_fullStr |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
title_full_unstemmed |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
title_sort |
Development of a Nanoscale-Sensitive DNA Sensor using Functionalized Graphene Substrates and Fluorescence Lifetime Characterization |
author |
Adão, Ricardo Miguel Ribeiro |
author_facet |
Adão, Ricardo Miguel Ribeiro |
author_role |
author |
dc.contributor.none.fl_str_mv |
Silva, Ana G. Nieder, Jana B. RUN |
dc.contributor.author.fl_str_mv |
Adão, Ricardo Miguel Ribeiro |
dc.subject.por.fl_str_mv |
Graphene nanoscale distance sensing fluorescence lifetime FLIM energy transfer DNA biosensing MATLAB Grafeno medição de distância à nanoescala tempo de vida de fluorescência FLIM transferência de energia biosensor de ADN MATLAB Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
topic |
Graphene nanoscale distance sensing fluorescence lifetime FLIM energy transfer DNA biosensing MATLAB Grafeno medição de distância à nanoescala tempo de vida de fluorescência FLIM transferência de energia biosensor de ADN MATLAB Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias |
description |
Fluorescence Lifetime Image Microscopy (FLIM) of nanostructured graphene substrates was used to measure the recently observed nanoscale distance-scaling of the fluorescence lifetime of dyes located in the vicinity of graphene sheets, at distances up to about 30 nm. The results were compared with a Resonant Energy Transfer (RET) theoretical model and used to establish an experimental fluorescence lifetime-to-nanoscale distance conversion function. In the following, this nano-optical relation was used for the design of a Deoxyribonucleic Acid (DNA) biosensor. Graphene was functionalized with fluorescently labeled DNA molecular beacons that unfold during hybridization with complementary DNA, and thereby change the distance of the fluorescent dye from the graphene surface. The spatial distribution of DNA molecular beacons binding to the surface of a graphene flake was studied, as well as the temporal kinetics of the hybridization reaction using time-lapse FLIM measurements. The results showed a vertical ascent of a fluorescent label relative to the graphene surface with a distance extension that is in accordance with the expected molecular length of the specific DNA sequence used. Interestingly, an intermediate state associated to a distance of a few nanometers was identified with a lifespan of about 85 minutes. The developed graphene-based DNA sensor was shown to enable optical detection of nanoscale distances in liquid media. These findings indicate that the fluorescence lifetime-based detection coupled with nanoscale interaction effects may find applications in various biosensing applications such as health and food-quality tracing. For the processing of FLIM data, several fluorescence lifetime calculation algorithms were compared and integrated into a specially designed and implemented analysis software toolbox in MATLAB. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-09-27 2017-09-27T00:00:00Z 2018-10-02T00:30:13Z |
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/10362/23795 |
url |
http://hdl.handle.net/10362/23795 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
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embargoedAccess |
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application/pdf |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799137905907597312 |