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

Area of Science:

• Forensic Science
• Analytical Chemistry
• Biotechnology

Background:

• Short tandem repeat (STR) analysis is crucial for human identification.
• Microchip-based platforms offer portability and reduced sample volume for genetic analysis.
• Current multicolor detection methods for STR analysis on microchips face limitations.

Purpose of the Study:

• To develop and demonstrate a single-filter, single-detector acousto-optic tunable filter (AOTF) system for multicolor fluorescence detection.
• To perform four- and five-color STR analysis on glass microchips.
• To integrate state-of-the-art multicolor STR kits with microchip technology.

Main Methods:

• Utilized an acousto-optic tunable filter (AOTF) for multicolor fluorescence detection.
• Employed AmpFlSTR Profiler Plus, COfiler (four-color), and Identifiler (five-color) STR kits.
• Generated matrix files and processed raw data to correct for spectral overlap.

Main Results:

• Successfully performed four- and five-color STR analysis on glass microchips using a single AOTF and detector.
• Demonstrated the first on-chip five-color genotyping application.
• Achieved spectral overlap correction for accurate STR analysis.

Conclusions:

• The AOTF system provides a robust and efficient method for multicolor STR analysis on microchips.
• This technology bridges the gap between portable microchip devices and advanced STR genotyping capabilities.
• The developed system enhances the potential for field-deployable forensic genetic analysis.