We demonstrate theoretically and experimentally that one-, two-, and three-photon excited fluorescence from dye molecules in spherical microcavities has an asymmetrical angular distribution and is enhanced in the backward direction. The enhancement ratios (of intensities at 180 degrees and 90 degrees ) are 9, 5, and 1.8 for three-, two-, and one-photon excitation, respectively. Even larger ratios are expected for microspheres with an index of refraction larger than that used in the experiments. Because of the reciprocity principle and concentration of the incident wave inside particles, the backward enhancement is expected to occur even with nonspherical particles.

Songbirds have been evolved into models of choice for the study of the cerebral underpinnings of vocal communication. Nevertheless, there is still a need for in vivo methods allowing the real-time monitoring of brain activity. Functional Magnetic Resonance Imaging (fMRI) has been applied in anesthetized intact songbirds. It relies on blood oxygen level-dependent (BOLD) contrast revealing hemodynamic changes. Non-invasive near-infrared spectroscopy (NIRS) is based on the weak absorption of near-infrared light by biological tissues. Time-resolved femtosecond white laser NIRS is a new probing method using real-time spectral measurements which give access to the local variation of absorbing chromophores such as hemoglobins. In this study, we test the efficiency of our time-resolved NIRS device in monitoring physiological hemodynamic brain responses in a songbird, the zebra finch (Taeniopygia guttata), using a hypercapnia event (7% inhaled CO(2)). The results are compared to those obtained using BOLD fMRI. The NIRS measurements clearly demonstrate that during hypercapnia the blood oxygen saturation level increases (increase in local concentration of oxyhemoglobin, decrease in deoxyhemoglobin concentration and total hemoglobin concentration). Our results provide the first correlation in songbirds of the variations in total hemoglobin and oxygen saturation level obtained from NIRS with local BOLD signal variations.

By assessing the cerebral blood volume and the hemoglobin oxygen saturation level, near-infrared spectroscopy (NIRS) probes brain oxygenation, which reflects cerebral activity. To develop a noninvasive method monitoring the brain of a songbird, we use an original NIRS device, i.e., a white laser coupled with an ultrafast spectrotemporal detector of optical signals without wavelength scanning. We perform in vivo measurements of the absorption coefficient and the reduced scattering coefficient of the caudal nidopallium area of the head of a songbird (the zebra finch).

The grants should have included the following note: 'These experiments were supported by the Program 'Emergence' of the Région Rhône-Alpes and the Agence Nationale de la Recherche (Project 'Birds' voices, ANR-06-BLAN-0293-01)'.