Fluorescence-Encoded Infrared Spectroscopy: Ultrafast Vibrational Spectroscopy on Small Ensembles of Molecules in...

Fluorescence-encoded infrared (FEIR) spectroscopy is an ultrafast technique that uses a visible pulse to up-convert information about IR-driven vibrations into a fluorescent electronic population. Here we present an updated experimental approach to FEIR that achieves high sensitivity through confocal microscopy, high repetition rate excitation, and single-photon counting. We demonstrate the sensitivity of our experiment by measuring ultrafast vibrational transients and Fourier transform spectra of increasingly dilute solutions of a coumarin dye. We collect high-quality data at 40 μM (∼2 orders of magnitude below the limit for conventional IR) and make measurements down to the 10-100 nM range (∼5 orders of magnitude) before background signals become overwhelming. At 10 nM we measure the average number of molecules in the focal volume to be ∼20 using fluorescence correlation spectroscopy. This level of sensitivity opens up the possibility of performing fluctuation correlation vibrational spectroscopy or-with further improvement-single-molecule measurements.