Bio-assay based on single molecule fluorescence detection in microfluidic channels.
A rapid bioassay is described based on the detection of colocalized fluorescent DNA probes bound to DNA targets in a pressure-driven solution flowing through a planar microfluidic channel. By employing total internal reflection excitation of the fluorescent probes and illumination of almost the entire flow channel, single fluorescent molecules can be efficiently detected leading to the rapid analysis of nearly the entire solution flowed through the device. Cross-correlation between images obtained from two spectrally distinct probes is used to determine the target concentration and efficiently reduces the number of false positives. The rapid analysis of DNA targets in the low pM range in less than a minute is demonstrated.
Hollars
Christopher W
CW
Lawrence Livermore National Laboratory, Chemistry and Materials Science, Livermore, CA, 94550, USA. chollars@llnl.gov
Puls
Jana
J
Bakajin
Olgica
O
Olsan
Brad
B
Talley
Chad E
CE
Lane
Stephen M
SM
Huser
Thomas
T
D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000379:methods*; D009693:Nucleic Acid Hybridization
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA