Multicolor fluorescence detection on an electrophoretic microdevice using an acoustooptic tunable filter

Anal Chem. 2006 Aug 1;78(15):5590-6. doi: 10.1021/ac0607358.

Abstract

An acoustooptic tunable filter (AOTF) is used to detect multiple fluorescent signals on a fluidic microdevice. A confocal laser-induced fluorescence detection setup is used to excite fluorescent dyes in glass microchannels, presenting a streamlined and robust detection system consisting of the narrow-bandwidth AO filter and a single photodetector. The flexibility of the filter is demonstrated by alternating between wavelengths for precise microchannel alignment and sweeping through a range of wavelengths for preliminary spectral characterization of subnanoliter probe volumes of target analytes. The AOTF is also coupled with an electrophoretic separation for the multicolor detection of PCR-amplified DNA against a labeled sizing standard, the discrimination of multiple amplicons overlapped in time, and the identification of amplified biowarfare agents in a fluorescent spiking experiment. Finally, to demonstrate the multicolor capability of the system, 19-wavelength detection is performed during the separation of a three-dye sample mixture.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acoustics
  • Electrophoresis, Microchip / instrumentation
  • Electrophoresis, Microchip / methods*
  • Fluorescence
  • Fluorescent Dyes / analysis*
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Fluorescent Dyes