AOTF-based multicolor fluorescence detection for short tandem repeat (STR) analysis in an electrophoretic microdevice

An acousto-optic tunable filter (AOTF) has been used to perform multicolor fluorescence detection for four and five-color short tandem repeat (STR) analysis on glass microchips. Matrix files were initially generated by collecting and comparing the laser-induced fluorescence emission of the labels specific to a particular STR kit, and raw data was processed to remove spectral overlap. The AmpFlSTR® kits used in this work include Profiler Plus™ and COfiler™, which are four-color kits used in tandem to address the core STR loci, as well as the five-color Identifiler™ kit, which contains each of the loci. In contrast to previous reports on multicolor detection for STR analysis on microchips, this detection system is characterized by a single filter and detector, and reports the first five-color genotyping application on-chip. This capability matches the portability and reduced scale of the microchip with the state-of-the-art in multicolor STR analysis kits.