Fluorescence-lifetime identification of biological agents using deep ultraviolet light-emitting diodes

Recently developed deep-UV light-emitting diodes (LEDs) are already used in prototype fluorescence sensors for detection of hazardous biological agents. However, increasing of the sensor ability of discrimination against common interferents requires further development of measurement technique. In particular, LED-based fluorescence lifetime measurements are to be considered as a technique supplementary to fluorescence spectral and excitation measurements. Here we report on application of UVTOP® series deep-UV LEDs developed by Sensor Electronic Technology, Inc. for real-time measurements of fluorescence lifetime in the frequency domain. LEDs with the wavelengths of 280 nm (targeted to protein excitation) and 340 nm (for excitation of coenzymes NADH and flavins) were used. The output of the LEDs was harmonically modulated at frequencies up to 100 MHz and fluorescence lifetime on the nanosecond and subnanosecond scale was estimated by measuring the phase angle of the fluorescence signal in respect of the LED output. Dual-wavelength LED-based phase-resolved measurement technique was tested for discrimination of B. globigii against a variety of interferents such as diesel fuel, paper, cotton, dust, etc. We conclude that fluorescence phase measurements have potential to improve the discrimination ability of the "detect-to-warn" optical bioparticle sensors.