Ultrarapid generation of femtoliter microfluidic droplets for single-molecule-counting immunoassays.
ACS Nano
; 7(7): 5955-64, 2013 Jul 23.
Article
em En
| MEDLINE
| ID: mdl-23805985
ABSTRACT
We report a microfluidic droplet-based approach enabling the measurement of chemical reactions of individual enzyme molecules and its application to a single-molecule-counting immunoassay. A microfluidic device is used to generate and manipulate <10 fL droplets at rates of up to 1.3 × 10(6) per second, about 2 orders of magnitude faster than has previously been reported. The femtodroplets produced with this device can be used to encapsulate single biomolecular complexes tagged with a reporter enzyme; their small volume enables the fluorescent product of a single enzyme molecule to be detected within 10 min of on-chip incubation. Our prototype system is validated by detection of a biomarker for prostate cancer in buffer, down to a concentration of 46 fM. This work demonstrates a highly flexible and sensitive diagnostic platform that exploits extremely high-speed generation of monodisperse femtoliter droplets for the counting of individual analyte molecules.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Neoplasias da Próstata
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Imunoensaio
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Técnicas Biossensoriais
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Antígeno Prostático Específico
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Técnicas Analíticas Microfluídicas
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Microquímica
Limite:
Humans
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Male
Idioma:
En
Revista:
ACS Nano
Ano de publicação:
2013
Tipo de documento:
Article