Time-resolved Förster-resonance-energy-transfer DNA assay on an active CMOS microarray.
We present an active oligonucleotide microarray platform for time-resolved Förster-resonance-energy-transfer (TR-FRET) assays. In these assays, immobilized probe is labeled with a donor fluorophore and analyte target is labeled with a fluorescence quencher. Changes in the fluorescence decay lifetime of the donor are measured to determine the extent of hybridization. In this work, we demonstrate that TR-FRET assays have reduced sensitivity to variances in probe surface density compared with standard fluorescence-based microarray assays. Use of an active array substrate, fabricated in a standard complementary metal-oxide-semiconductor (CMOS) process, provides the additional benefits of reduced system complexity and cost. The array consists of 4096 independent single-photon avalanche diode (SPAD) pixel sites and features on-chip time-to-digital conversion. We demonstrate the functionality of our system by measuring a DNA target concentration series using TR-FRET with semiconductor quantum dot donors.
Department of Electrical Engineering, Bioelectronic Systems Laboratory, Columbia University, NY 10027, USA. des@cisl.columbia.edu
David Eric
DE
Schwartz
Ping
P
Gong
Kenneth L
KL
Shepard
D004247:DNA / Q000032:analysis*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D045663:Quantum Dots; D012666:Semiconductors
D004247:DNA