期刊
ACS NANO
卷 7, 期 7, 页码 5955-5964出版社
AMER CHEMICAL SOC
DOI: 10.1021/nn401661d
关键词
microfluidics; fast droplet generation; single-enzyme analysis; digital immunoassay
类别
资金
- Engineering and Physical Sciences Research Council
- European Commission (TheraEDGE Network) [FP7-216027]
- EU
- Isaac Newton Trust
- BBSRC [BB/F00219X/1, BB/D014964/1] Funding Source: UKRI
- EPSRC [EP/H046593/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/D014964/1, BB/F00219X/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [EP/H046593/1] Funding Source: researchfish
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 x 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.
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