A simple and highly sensitive spectroscopic fluorescence-detection system for multi-channel plastic-microchip electrophoresis based on side-entry laser-beam zigzag irradiation

Lab Chip. 2017 Jun 27;17(13):2235-2242. doi: 10.1039/c7lc00448f.

Abstract

A five-color fluorescence-detection system for eight-channel plastic-microchip electrophoresis was developed. In the eight channels (with effective electrophoretic lengths of 10 cm), single-stranded DNA fragments were separated (with single-base resolution up to 300 bases within 10 min), and seventeen-loci STR genotyping for forensic human identification was successfully demonstrated. In the system, a side-entry laser beam is passed through the eight channels (eight A channels), with alternately arrayed seven sacrificial channels (seven B channels), by a technique called "side-entry laser-beam zigzag irradiation." Laser-induced fluorescence from the eight A channels and Raman-scattered light from the seven B channels are then simultaneously, uniformly, and spectroscopically detected, in the direction perpendicular to the channel array plane, through a transmission grating and a CCD camera. The system is therefore simple and highly sensitive. Because the microchip is fabricated by plastic-injection molding, it is inexpensive and disposable and thus suitable for actual use in various fields.

MeSH terms

  • DNA / analysis
  • DNA / chemistry
  • DNA / genetics
  • Electrophoresis, Microchip / instrumentation*
  • Forensic Genetics / instrumentation*
  • Forensic Genetics / methods
  • Humans
  • Microsatellite Repeats / genetics
  • Sequence Analysis, DNA / instrumentation*
  • Sequence Analysis, DNA / methods
  • Spectrometry, Fluorescence / instrumentation*

Substances

  • DNA