FLUORESCENCE LIFETIME MEASUREMENTS USING THERMAL SOURCE

Evita Spalinskaitė1, Vytautas Žičkus1, 2, Zigmas Balevičius1

1 Plasmonics and Nanophotonics Lab., Department of Laser Technologies, Center for Physical Sciences and Technology, Saulėtekio av. 3, LT-10257 Vilnius, Lithuania

2 School of Physics and Astronomy, University of Glasgow, Glasgow G12 8QQ, United Kingdom

[email protected]

Fluorescence lifetime measurements are widely used in exploring various biological environments, revealing changes in pH, temperature, and viscosity among other parameters [1]. Measurements are typically performed with time-correlated single photon counting (TCSPC) methods, which are limited to 100’s of picoseconds lifetime resolution due to limitations of electrical hardware [2]. Recently, Fluorescence lifetime Hong-Ou-Mandel sensing (FLHOM) method [2], where temporal resolution is limited by optical properties of light, overcomes the limitations of standard technique. Here, we introduce a low-cost version of the FLHOM method, by replacing a coherent reference source with a thermal source.

We used Allura Red fluorescent dye, with a nominal lifetime of ~1.6 ps [2], as a reference source to probe the lifetime of a 4-DASPI dye in a Mach-Zehnder interferometer set up. Secondly, we deconvolved the measured second-order correlation function and recovered fluorescence lifetime of 5.6 ps, in agreement with previous findings [2]. We measured a reduced interference visibility around 6.5% due to intrinsically bunched nature of thermal light. In conclusion, integrating fluorescent dye as a reference thermal source reduces the complexity and cost of the experimental system. Despite marginal reduction in achievable lifetime resolution, the method opens opportunities to measure ultrashort fluorescence lifetime.


[1] Datta R., Heaster T. M., Sharick J. T., et al., Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications, J. Biomed. Opt. 25(7), 071203 (2020)

[2] Lyons, A., Zickus, V., Álvarez-Mendoza, R. et al., Fluorescence lifetime Hong-Ou-Mandel sensing. Nat Commun 14, 8005 (2023)