Chemiluminescence and Bioluminescence

Kricka, Larry J.

doi:10.1021/a19999022

Cited by 39 publications

(33 citation statements)

References 41 publications

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“…The cantilever design enables functionalization of individual sensors in distinct reservoirs. Each sensor has a 50 ϫ 50-m 2 sensing region doped with Ϸ10 15 atoms per cm 3 and is covered with an Ϸ2-nm layer of chemically grown oxide. Both the low doping level and extremely thin insulator thickness enhance the surface potential sensitivity of the sensors (16).…”

Section: Methodsmentioning

confidence: 99%

“…2). In the methods used most routinely, the physical nature of the readout requires the attachment of reporter molecules such as fluorescent, chemiluminescent, redox, or radioactive labels (1,3,4). Although labeldependent methods achieve the highest sensitivities (5-7), eliminating the labeling steps has the advantage of simplifying the readout and increasing the speed and ease of nucleic acid assays, which is especially desirable for characterizing infectious agents, scoring sequence polymorphisms and genotypes, and measuring mRNA levels during expression profiling.…”

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confidence: 99%

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Electronic detection of DNA by its intrinsic molecular charge

Fritz

Cooper

Gaudet

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

428

320

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We report the selective and real-time detection of label-free DNA using an electronic readout. Microfabricated silicon field-effect sensors were used to directly monitor the increase in surface charge when DNA hybridizes on the sensor surface. The electrostatic immobilization of probe DNA on a positively charged poly-L-lysine layer allows hybridization at low ionic strength where field-effect sensing is most sensitive. Nanomolar DNA concentrations can be detected within minutes, and a single base mismatch within 12-mer oligonucleotides can be distinguished by using a differential detection technique with two sensors in parallel. The sensors were fabricated by standard silicon microtechnology and show promise for future electronic DNA arrays and rapid characterization of nucleic acid samples. This approach demonstrates the most direct and simple translation of genetic information to microelectronics. Awide range of techniques for detecting nucleic acids is based on their hybridization to DNA probes on a solid surface (ref. 1, entire issue, and ref. 2). In the methods used most routinely, the physical nature of the readout requires the attachment of reporter molecules such as fluorescent, chemiluminescent, redox, or radioactive labels (1, 3, 4). Although labeldependent methods achieve the highest sensitivities (5-7), eliminating the labeling steps has the advantage of simplifying the readout and increasing the speed and ease of nucleic acid assays, which is especially desirable for characterizing infectious agents, scoring sequence polymorphisms and genotypes, and measuring mRNA levels during expression profiling. The development of label-independent methods that can monitor hybridization in real time and that are simple and scalable is still in its infancy (8-11). Here we describe a label-free method for electronically detecting DNA by its intrinsic molecular charge using microfabricated field-effect sensors.The field-effect sensor is based on an electrolyte-insulator-silicon (EIS) structure. Variations in the insulator-electrolyte surface potential, which arise from the binding of charged molecules (e.g., nucleic acids) to the insulator surface ( Fig. 1 a and b), modify the charge distribution in the silicon below the electrolyte. Surface charge and surface potential at this interface are related according to the Grahame equation (12). The surface potential can be measured by changes in conductivity (13) or capacitance (14) in the silicon part of the EIS structure. We have chosen to measure the capacitance, because it requires only one electrical connection to the silicon. The measured capacitance between the silicon and counter electrode in the electrolyte solution is dominated by the insulator capacitance and the capacitance of the charge-depleted region in the silicon. These two capacitances appear in series, and only the silicon depletion capacitance is modulated by the insulator surface potential. The capacitance-versus-voltage dependence of EIS structures (15) is similar to that of metal-oxide-semiconducto...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Electronic detection of DNA by its intrinsic molecular charge

Fritz

Cooper

Gaudet

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

428

320

View full text Add to dashboard Cite

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“…Chemiluminescence (CL) has been shown to be a highly sensitive method for the detection in conventional capillary electrophoresis (CE) [16,17]. CL can be defined as the emission of light (ultraviolet, visible or infrared) from a molecule or atom in an electronically excited state produced by a chemical reaction [17].…”

Section: Introductionmentioning

confidence: 99%

“…CL can be defined as the emission of light (ultraviolet, visible or infrared) from a molecule or atom in an electronically excited state produced by a chemical reaction [17]. It needs no light sources, and thus significantly reduces noise and eliminates Rayleigh and Raman scattering, allowing photon detectors to be operated at high gains to improve the ratio of signal to noise.…”

Section: Introductionmentioning

confidence: 99%

Integration of a flow-type chemiluminescence detector on a glass electrophoresis chip

Lin

Uchiyama

et al. 2004

Talanta

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“…The first diagnostic tests based on PCR were quite cumbersome and not amenable to large-scale screening methods (2). Several post-PCR steps, such as restriction enzyme analysis, agarose gel electrophoresis or heterogeneous hybridization assays were needed to confirm the identity of the PCR product (3). Recently, the development of new fluorescent techniques has, however, led to novel assay formats that greatly simplify the protocols used for the detection of specific nucleic acid sequences.…”

Section: Introductionmentioning

confidence: 99%

A new label technology for the detection of specific polymerase chain reaction products in a closed tube

Nurmi

Ylikoski

Soukka

et al. 2000

Nucleic Acids Research

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A novel signal generation principle suitable for real time and end-point detection of specific PCR products in a closed tube is described. Linear DNA probes were labeled at their 5′-ends with a stable, fluorescent terbium chelate. The fluorescence intensity of this chelate is lower when it is coupled to single-stranded DNA than when the chelate is free in solution. The synthesized probes were used in the real time monitoring of PCR using a prototype instrument that consisted of a fluorometer coupled to a thermal cycler. When the probe anneals to a complementary target amplicon, the 5′→3′ exonucleolytic activity of DNA polymerase detaches the label from the probe. This results in an enhanced terbium fluorescence signal. Since terbium has a long excited state lifetime, its fluorescence can be measured in a time-resolved manner, which results in a low background fluorescence and a 1000-fold signal amplification. The detection method is quantitative over an extremely wide linear range (at least 10-10 7 initial template molecules). The label strategy can easily be combined with existing label technologies, such as TaqMan 5′-exonuclease assays, in order to carry out multiplex assays that do not suffer from overlapping emission peaks of the fluorophores.

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Chemiluminescence and Bioluminescence

Cited by 39 publications

References 41 publications

Electronic detection of DNA by its intrinsic molecular charge

Electronic detection of DNA by its intrinsic molecular charge

Integration of a flow-type chemiluminescence detector on a glass electrophoresis chip

A new label technology for the detection of specific polymerase chain reaction products in a closed tube

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