We describe the luminescence spectral properties of CdS nanoparticles with multiphoton excitation. Three types of CdS nanoparticles were examined which were a CdS/dendrimer composite which displays high anisotropy, Cd 2+ -enriched nanoparticles which display two emission maxima, and polyphosphate-stabilized nanoparticles which display long wavelength emission. Illumination with long wavelengths near 700-790 nm resulted in two-photon excitation. Essentially the same emission spectra and intensity decays were observed with one-photon and two-photon excitation. Comparison with fluorescein indicates the NPs display large two-photon cross sections near 100 GM. The CdS/dendrimer and Cd 2+ -enriched CdS nanoparticles displayed large anisotropy values with two-photon excitation, substantially larger than with one-photon excitation. It appears that semiconductor nanoparticles are comparable to organic fluorophores which display the same spectral properties with one-photon and two-photon excitation.

Phytofluors are highly fluorescent proteins in which the chromophore in a phytochrome is replaced with phycoerythrobilin (PEB), the pigment precursor of the cyanobacterial light harvesting protein phycoerythrin. We examined the fluorescence spectra of the N-terminal region of the cyanobacterial phytochrome 1 from cyanobacterium Synechocystis sp. Pcc 6803 bound to PEB. This protein, Cph1(N514)-PEB, displayed a good two-photon cross-section of 20-30 GM for excitation at 792 nm. This phytofluor also exhibits a high fundamental anisotropy at most practical two-photon excitation (2PE) wavelengths from 700 to 900 nm. Identical lifetimes and correlation times with one and 2PE indicates that the phytofluor is not adversely affected by the intensities needed for 2PE. The one-photon absorption extends well beyond the absorption spectrum and even beyond the emission spectrum to 700 nm. The phytofluor thus appears to be a suitable probe for 2PE and/or cellular imaging.