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1.
Anal Chem ; 83(3): 888-95, 2011 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-21214189

RESUMO

We demonstrate the temperature mediated applications of a previously proposed novel localized dielectric heating method on the surface of dual purpose silicon field effect transistor (FET) sensor-heaters and perform modeling and characterization of the underlying mechanisms. The FETs are first shown to operate as electrical sensors via sensitivity to changes in pH in ionic fluids. The same devices are then demonstrated as highly localized heaters via investigation of experimental heating profiles and comparison to simulation results. These results offer further insight into the heating mechanism and help determine the spatial resolution of the technique. Two important biosensor platform applications spanning different temperature ranges are then demonstrated: a localized heat-mediated DNA exchange reaction and a method for dense selective functionalization of probe molecules via the heat catalyzed complete desorption and reattachment of chemical functionalization to the transistor surfaces. Our results show that the use of silicon transistors can be extended beyond electrical switching and field-effect sensing to performing localized temperature controlled chemical reactions on the transistor itself.


Assuntos
Temperatura Alta , Silício/química , Transistores Eletrônicos , Catálise , Microscopia Eletrônica de Varredura , Propriedades de Superfície
2.
Lab Chip ; 9(19): 2789-95, 2009 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-19967115

RESUMO

We demonstrate electrically addressable localized heating in fluid at the dielectric surface of silicon-on-insulator field-effect transistors via radio-frequency Joule heating of mobile ions in the Debye layer. Measurement of fluid temperatures in close vicinity to surfaces poses a challenge due to the localized nature of the temperature profile. To address this, we developed a localized thermometry technique based on the fluorescence decay rate of covalently attached fluorophores to extract the temperature within 2 nm of any oxide surface. We demonstrate precise spatial control of voltage dependent temperature profiles on the transistor surfaces. Our results introduce a new dimension to present sensing systems by enabling dual purpose silicon transistor-heaters that serve both as field effect sensors as well as temperature controllers that could perform localized bio-chemical reactions in Lab on Chip applications.


Assuntos
Temperatura Alta , Silício/química , Transistores Eletrônicos , Corantes Fluorescentes/química , Dispositivos Lab-On-A-Chip , Micro-Ondas , Propriedades de Superfície
3.
Anal Biochem ; 308(2): 358-63, 2002 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-12419350

RESUMO

S-Adenosyl-L-methionine (AdoMet) which is biologically synthesized by AdoMet synthetase bears an S configuration at the sulfur atom. The chiral sulfonium spontaneously racemizes to form a mixture of S and R isomers of AdoMet under physiological conditions or normal storage conditions. The chirality of AdoMet greatly affects its activity; the R isomer is not accepted as a substrate for AdoMet-dependent methyltransferases. We report a stereospecific colorimetric assay for (S,S)-adenosylmethionine quantification based on an enzyme-coupled reaction in which (S,S)-AdoMet reacts with 2-nitro-5-thiobenzoic acid to form AdoHcy and 2-nitro-5-methylthiobenzoic acid. The transformation is catalyzed by recombinant human thiopurine S-methyltransferase (TPMT, EC 2.1.1.67) and is associated with a large spectral change at 410 nm. Accumulation of the S-adenosylhomocysteine (AdoHcy) product, a feedback inhibitor of TPMT, slows the assay. AdoHcy nucleosidase (EC 3.2.2.9) irreversibly cleaves AdoHcy to adenine and S-ribosylhomocysteine, significantly shortening the assay time to less than 10 min. The assay is linear from 5 to at least 60 microM (S,S)-AdoMet.


Assuntos
Metiltransferases/metabolismo , Polilisina/análogos & derivados , S-Adenosilmetionina/análise , Compostos de Sulfidrila/metabolismo , Calorimetria , Escherichia coli/enzimologia , Humanos , Indicadores e Reagentes , Cinética , Metilação , Plasmídeos , Proteínas Recombinantes/isolamento & purificação , Estereoisomerismo , Trinitrobenzenos/farmacologia
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