Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells
Autor(a) principal: | |
---|---|
Data de Publicação: | 2015 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/35343 |
Resumo: | An important step toward understanding interactions between nanoparticles (NPs) and bacteria is the ability to directly observe NPs interacting with bacterial cells. NPbacteria mixtures typical in nanomedicine, however, are not yet amendable for direct imaging in solution. Instead, evidence of NPcell interactions must be preserved in derivative (usually dried) samples to be subsequently revealed in high-resolution images, e.g., via scanning electron microscopy (SEM). Here, this concept is realized for a mixed suspension of model NPs and Staphylococcus aureus bacteria. First, protocols for analyzing the relative colloidal stabilities of NPs and bacteria are developed and validated based on systematic centrifugation and comparison of colony forming unit (CFU) counting and optical density (OD) measurements. Rate-dependence of centrifugation efficiency for each component suggests differential sedimentation at a specific predicted rate as an effective method for removing free NPs after co-incubation; the remaining fraction comprises bacteria with any associated NPs and can be examined, e.g., by SEM, for evidence of NPbacteria interactions. These analytical protocols, validated by systematic control experiments and high-resolution SEM imaging, should be generally applicable for investigating NPbacteria interactions. |
id |
RCAP_808f28277eb1f3d45965c42528f77489 |
---|---|
oai_identifier_str |
oai:repositorium.sdum.uminho.pt:1822/35343 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cellsEngenharia e Tecnologia::NanotecnologiaScience & TechnologyAn important step toward understanding interactions between nanoparticles (NPs) and bacteria is the ability to directly observe NPs interacting with bacterial cells. NPbacteria mixtures typical in nanomedicine, however, are not yet amendable for direct imaging in solution. Instead, evidence of NPcell interactions must be preserved in derivative (usually dried) samples to be subsequently revealed in high-resolution images, e.g., via scanning electron microscopy (SEM). Here, this concept is realized for a mixed suspension of model NPs and Staphylococcus aureus bacteria. First, protocols for analyzing the relative colloidal stabilities of NPs and bacteria are developed and validated based on systematic centrifugation and comparison of colony forming unit (CFU) counting and optical density (OD) measurements. Rate-dependence of centrifugation efficiency for each component suggests differential sedimentation at a specific predicted rate as an effective method for removing free NPs after co-incubation; the remaining fraction comprises bacteria with any associated NPs and can be examined, e.g., by SEM, for evidence of NPbacteria interactions. These analytical protocols, validated by systematic control experiments and high-resolution SEM imaging, should be generally applicable for investigating NPbacteria interactions.financial support from the following sources: grant SFRH/BPD/47693/2008 from the Portuguese Foundation for Science and Technology (FCT); FCT Strategic Project PEst-OE/EQB/LA0023/2013; project “BioHealth Biotechnology and Bioengineering approaches to improve health quality”, Ref. NORTE-07-0124-FEDER-000027, cofunded by the Programa Operacional Regional do Norte (ON.2−O Novo Norte), QREN, FEDER; project “Consolidating Research Expertise and Resources on Cellular and Molecular Biotechnology at CEB/IBB”, ref. FCOMP-01-0124-FEDER- 027462.American Chemical SocietyUniversidade do MinhoSousa, CláudiaSequeira, DianaKolen´ko, Yury V.Pinto, Inês M.Petrovykh, D. Y.2015-032015-03-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/35343engSousa, Cláudia; Sequeira, Diana; Kolen'ko, Yury V.; Pinto, Inês M.; Petrovykh, D. Y., Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells. Analytical Chemistry, 87(9), 4641-4648, 20150003-27001520-688210.1021/ac503835a25764323http://pubs.acs.org/doi/abs/10.1021/ac503835ainfo: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:RCAAP2023-07-21T12:09:14Zoai:repositorium.sdum.uminho.pt:1822/35343Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:00:36.531653Repositório 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 |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
title |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
spellingShingle |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells Sousa, Cláudia Engenharia e Tecnologia::Nanotecnologia Science & Technology |
title_short |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
title_full |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
title_fullStr |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
title_full_unstemmed |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
title_sort |
Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells |
author |
Sousa, Cláudia |
author_facet |
Sousa, Cláudia Sequeira, Diana Kolen´ko, Yury V. Pinto, Inês M. Petrovykh, D. Y. |
author_role |
author |
author2 |
Sequeira, Diana Kolen´ko, Yury V. Pinto, Inês M. Petrovykh, D. Y. |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Sousa, Cláudia Sequeira, Diana Kolen´ko, Yury V. Pinto, Inês M. Petrovykh, D. Y. |
dc.subject.por.fl_str_mv |
Engenharia e Tecnologia::Nanotecnologia Science & Technology |
topic |
Engenharia e Tecnologia::Nanotecnologia Science & Technology |
description |
An important step toward understanding interactions between nanoparticles (NPs) and bacteria is the ability to directly observe NPs interacting with bacterial cells. NPbacteria mixtures typical in nanomedicine, however, are not yet amendable for direct imaging in solution. Instead, evidence of NPcell interactions must be preserved in derivative (usually dried) samples to be subsequently revealed in high-resolution images, e.g., via scanning electron microscopy (SEM). Here, this concept is realized for a mixed suspension of model NPs and Staphylococcus aureus bacteria. First, protocols for analyzing the relative colloidal stabilities of NPs and bacteria are developed and validated based on systematic centrifugation and comparison of colony forming unit (CFU) counting and optical density (OD) measurements. Rate-dependence of centrifugation efficiency for each component suggests differential sedimentation at a specific predicted rate as an effective method for removing free NPs after co-incubation; the remaining fraction comprises bacteria with any associated NPs and can be examined, e.g., by SEM, for evidence of NPbacteria interactions. These analytical protocols, validated by systematic control experiments and high-resolution SEM imaging, should be generally applicable for investigating NPbacteria interactions. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-03 2015-03-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/35343 |
url |
http://hdl.handle.net/1822/35343 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Sousa, Cláudia; Sequeira, Diana; Kolen'ko, Yury V.; Pinto, Inês M.; Petrovykh, D. Y., Analytical protocols for separation and electron microscopy of nanoparticles interacting with bacterial cells. Analytical Chemistry, 87(9), 4641-4648, 2015 0003-2700 1520-6882 10.1021/ac503835a 25764323 http://pubs.acs.org/doi/abs/10.1021/ac503835a |
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.publisher.none.fl_str_mv |
American Chemical Society |
publisher.none.fl_str_mv |
American Chemical Society |
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_ |
1799132401846190080 |