Sub-nanomolar detection of thrombin activity on a microfluidic chip

Bioluminescence resonance energy transfer (BRET) is a form of Förster resonance energy transfer. BRET has been shown to support lower limits of detection than fluorescence resonance energy transfer (FRET) but, unlike FRET, has not been widely implemented on microfluidic devices for bioanalytical sen...

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Bibliographic Details
Published inBiomicrofluidics Vol. 8; no. 6; p. 064110
Main Authors Le, Nam Cao Hoai, Gel, Murat, Zhu, Yonggang, Wang, Jian, Dacres, Helen, Anderson, Alisha, Trowell, Stephen C
Format Journal Article
LanguageEnglish
Published United States American Institute of Physics 01.11.2014
AIP Publishing LLC
Subjects
Bioluminescence
Design optimization
Energy transfer
Fluorescence
Numerical aperture
Optical fibers
Organic chemistry
Regular
Response time
Thrombin
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Summary:Bioluminescence resonance energy transfer (BRET) is a form of Förster resonance energy transfer. BRET has been shown to support lower limits of detection than fluorescence resonance energy transfer (FRET) but, unlike FRET, has not been widely implemented on microfluidic devices for bioanalytical sensing. We recently reported a microscope-based microfluidic system for BRET-based biosensing, using a hybrid, high quantum-efficiency, form of BRET chemistry. This paper reports the first optical fiber-based system for BRET detection on a microfluidic chip, capable of quantifying photon emissions from the low quantum-efficiency BRET(2) system. We investigated the effects of varying core diameter and numerical aperture of optical fibers, as well as varying microfluidic channel design and measurement conditions. We optimized the set-up in order to maximize photon counts and minimize the response time. The optimized conditions supported measurement of thrombin activity, with a limit of detection of 20 pM, which is lower than the microscope-based system and more than 20 times lower than concentrations reported to occur in plasma clots.
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Author to whom correspondence should be addressed. Electronic mail: nam.le@csiro.au. Telephone: +61 3 9545 7971. Fax: +61 3 9545 2818.
ISSN:1932-1058
1932-1058
DOI:10.1063/1.4902908