Two-Photon Excitation Fluorescence Spectroscopy of Quantum Dots: Photophysical Properties and Application in Bioassays

The applications of quantum dots (QDs) in two-photon (2P) excitation applications demand reliable data about their 2P absorption (2PA) cross sections (σ2PA). In the present study, σ2PA values have been determined for a series of commercial colloidal CdSe/ZnS QDs and CdSeTe/ZnS QDs in aqueous media. For the first time for these QDs, the σ2PA values have been determined over a wide spectral range, that is, between 720 and 900 nm, and are compared to the extinction coefficient (ε) values obtained under one-photon (1P) excitation. Furthermore, we present a QD in combination with an organic dye in a biotin-streptavidin Förster resonance energy transfer bioassay under 1P and 2P excitation. The results for the bioassay under 2P excitation are compared to those obtained under 1P excitation. The results demonstrate that in the case of the 2P excitation, higher sensitivity can be achieved because of an improved signal-to-noise ratio.