Coupling confocal fluorescence detection and recirculating microfluidic control for single particle analysis in discrete nanoliter volumes.
The recent proliferation of platforms designed to handle arrays of nano- and picolitre volumes is in response to the need to perform biological assays on discrete entities, such as single cells. However, a critical challenge associated with this trend for in vitro compartmentalization is the need for highly sensitive, yet low-volume detection platforms. In this paper, we coupled confocal fluorescence detection with recirculating microfluidic control to perform single particle DNA assays within five nL chambers. The performance of this low-volume assay was shown to match that of traditional single molecule detection platforms. However, volume requirements per measurement were nearly 3 orders of magnitude less than conventional systems, enabling future integration with lab-on-a-chip systems that require discrete or digitalized sample processing.
Puleo
C M
CM
Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
Yeh
H C
HC
Liu
K J
KJ
Wang
T H
TH
D004247:DNA / Q000032:analysis*; D005914:Globins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008863:Microspheres; D013997:Time Factors
D005914:Globins; D004247:DNA