Abstract
We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays. The system was tested to measure cardiac marker Troponin I with a concentration of 200 ng/L in immunoassay. The signal-to-noise ratio was comparable to state-of-the-art Xenon flash lamp based unit with equal excitation energy and without overdriving the LED. We performed a comparative study of the flash lamp and the LED based system and discussed temporal, spatial, and spectral features of the LED excitation for time-resolved fluorimetry. Optimization of the suggested key parameters of the LED promises significant increase of the signal-to-noise ratio and hence of the sensitivity of immunoassay systems.
Highlights
Time-resolved fluorescence is a powerful tool that has been used in immunoassays detection to suppress short-lived background fluorescence and allow for high sensitivity [1]
We report on the design, development and investigation of an optical system based on UV light emitting diode (LED) excitation at 340 nm for time-resolved fluorescence detection of immunoassays
Including the light emitting diode (LED) small footprint, cheaper price, better efficiency and heat dissipation, we conclude that UV LEDs will likely soon replace Xenon flash lamp excitation in immunoassays time-resolved detection systems based on long lifetime fluorophores
Summary
Time-resolved fluorescence is a powerful tool that has been used in immunoassays detection to suppress short-lived background fluorescence and allow for high sensitivity [1] It relies on employing high-fluorescent fluorophores with long fluorescence lifetime and light sources emitting excitation pulses terminating prior to the gated measurement. LEDs at 340 nm with pulse energies comparable to flash lamps have only recently become available and to our knowledge, this is the first report of the design, development and characterization of a single emitter, 340 nm LED based time-resolved fluorescence system for immunoassays detection. We performed a comparative study of this high power LED with the Xenon flash lamp and discuss spatial, spectral and temporal features of LED illumination when used for time-resolved fluorescence measurements
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