Ultrasensitive time-resolved nanoliter volume fluorometry based on UV LEDs and a channel photomultiplier tube

Dayong Jin; Russell Connally; James Piper

doi:10.1117/12.590127

29 March 2005 Ultrasensitive time-resolved nanoliter volume fluorometry based on UV LEDs and a channel photomultiplier tube

Dayong Jin, Russell Connally, James Piper

Author Affiliations +

Proceedings Volume 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III; (2005) https://doi.org/10.1117/12.590127
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

A capillary fluorometer was constructed using a 2 mW, 365 nm ultraviolet (UV) light emitting diode (LED) as the excitation source and a new-generation high-gain (3×10⁸) channel photomultiplier tube. The use of a LED permitted rapid pulsing of the excitation source so that the instrument could be employed for time-resolved fluorescence (TRF) applications. A detection limit of ~2×10⁸ molecules of BHHT (4,4’-bis (1",1",1",2",2",3",3"-heptafluoro-4",6"-hexanedion-6"yl)-o-tephenyl)-Eu (III) were resolved within a 1.25 nanoliter volume at a S/N ratio of 3:1. Ultimate sensitivity of the system was compromised due to visible luminescence emitted by the UV LED, centred around 550 nm extending to > 700 nm and 2nd-order exponentially decaying with lifetimes of 40 μs and 490 μs.

Citation Download Citation

Dayong Jin, Russell Connally, and James Piper "Ultrasensitive time-resolved nanoliter volume fluorometry based on UV LEDs and a channel photomultiplier tube", Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); https://doi.org/10.1117/12.590127

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