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1.
Biosens Bioelectron ; 57: 162-70, 2014 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-24583687

RESUMO

This work deals with the design of a highly sensitive whole cell-based biosensor for heavy metal detection in liquid medium. The biosensor is constituted of a Love wave sensor coated with a polyelectrolyte multilayer (PEM). Escherichia coli bacteria are used as bioreceptors as their viscoelastic properties are influenced by toxic heavy metals. The acoustic sensor is constituted of a quartz substrate with interdigitated transducers and a SiO2 guiding layer. However, SiO2 shows some degradation when used in a saline medium. Mesoporous TiO2 presents good mechanical and chemical stability and offers a high active surface area. Then, the addition of a thin titania layer dip-coated onto the acoustic path of the sensor is proposed to overcome the silica degradation and to improve the mass effect sensitivity of the acoustic device. PEM and bacteria deposition, and heavy metal influence, are real time monitored through the resonance frequency variations of the acoustic device. The first polyelectrolyte layer is inserted through the titania mesoporosity, favouring rigid link of the PEM on the sensor and improving the device sensitivity. Also, the mesoporosity of surface increases the specific surface area which can be occupied and favors the formation of homogeneous PEM. It was found a frequency shift near -20±1 kHz for bacteria immobilization with titania film instead of -7±3 kHz with bare silica surface. The sensitivity is highlighted towards cadmium detection. Moreover, in this paper, particular attention is given to the immobilization of bacteria and to biosensor lifetime. Atomic Force Microscopy characterizations of the biosurface have been done for several weeks. They showed significant morphological differences depending on the bacterial life time. We noticed that the lifetime of the biosensor is longer in the case of using a mesoporous TiO2 layer.


Assuntos
Acústica/instrumentação , Biofilmes , Técnicas Biossensoriais/instrumentação , Escherichia/fisiologia , Metais Pesados/análise , Titânio/química , Biofilmes/crescimento & desenvolvimento , Cádmio/análise , Desenho de Equipamento , Escherichia/citologia , Metais Pesados/metabolismo , Porosidade , Sensibilidade e Especificidade , Transdutores
2.
Biosens Bioelectron ; 26(4): 1723-6, 2010 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-20810269

RESUMO

The present work deals with the development of a Love-wave bacteria-based sensor platform for the detection of heavy metals in liquid medium. The acoustic delay-line is inserted in an oscillation loop in order to record the resonance frequency in real-time. A Polydimethylsiloxane (PDMS) chip with a liquid chamber is maintained by pressure above the acoustic wave propagation path. Bacteria (Escherichia coli) were fixed as bioreceptors onto the sensitive surface of the sensor coated with a polyelectrolyte (PE) multilayer using a simple and efficient layer-by-layer (LbL) electrostatic self-assembly procedure. Poly(allylamine hydrochloride) (PAH cation) and poly(styrene sulfonate) (PSS anion) were alternatively deposited so that the strong attraction between oppositely charged polyelectrolytes resulted in the formation of a (PAH-PSS)(n)-PAH molecular multilayer. The real-time characterization of PE multilayer and bacteria deposition is based on the measurement of the resonance frequency perturbation due to mass loading during material deposition. Real-time response to various concentrations of cadmium (Cd(2+)) and mercury (Hg(2+)) has been investigated. A detection limit as low as 10(-12) mol/l has been achieved, above which the frequency increases gradually up to 10(-3) mol/l, after a delay of 60 s subsequent to their introduction onto bacterial cell-based biosensors. Beyond a 10(-3) mol/l a steep drop in frequency was observed. This response has been attributed to changes in viscoelastic properties, related to modifications in bacteria metabolism.


Assuntos
Técnicas Biossensoriais/métodos , Escherichia coli/efeitos dos fármacos , Metais Pesados/toxicidade , Acústica , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/estatística & dados numéricos , Cádmio/toxicidade , Células Imobilizadas , Meios de Cultura , Dimetilpolisiloxanos , Eletrólitos , Mercúrio/toxicidade , Poliaminas , Poliestirenos , Eletricidade Estática
3.
Biosens Bioelectron ; 18(5-6): 755-63, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12706589

RESUMO

Love wave sensors (ST-cut quartz substrate with interdigital transducers, SiO(2) guiding layer and sensitive coating) have been receiving a great deal of attention for a few years. Indeed, the wave coupled in a guiding layer confers a high gravimetric sensitivity and the shear horizontal (SH) polarization allows to work in liquid media. In this paper, an analytical method is proposed to calculate the Love wave phase velocity and the gravimetric sensitivity for a complete multilayer structure. This allows us to optimize the Love wave devices design in order to improve their gravimetric sensitivity in liquid media. As a model for virus or bacteria detection in liquids (drinking or bathing water, food em leader ) we design a model using M13 bacteriophage. The first step is the anti-M13 (AM13) monoclonal antibody grafting, on the device surface (SiO(2)). The second step is an immunoreaction in between the M13 bacteriophage and the AM13 antibody. The Love wave device allows to detect in real time the graft of the AM13 sensitive coating, as well as the immobilization of the M13 bacteriophages. With a pH change, the M13 bacteriophages can be removed from the sensor surface, in order to be numerated as plaque forming unit (pfu). Results on the sensitivity of Love waves are compared with similar immunological works with bulk acoustic wave devices, and demonstrate the high potentialities of Love waves sensors.


Assuntos
Acústica/instrumentação , Bacteriófago M13/isolamento & purificação , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Imunoensaio/instrumentação , Transdutores , Materiais Revestidos Biocompatíveis/síntese química , Materiais Revestidos Biocompatíveis/química , Desenho de Equipamento , Análise de Falha de Equipamento , Estudos de Viabilidade , Imunoensaio/métodos , Sistemas On-Line/instrumentação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Dióxido de Silício , Soluções/análise , Microbiologia da Água , Poluentes da Água/análise
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