We report for the first time the use of two photon fluorescence as detection method of affinity binding reactions. We use a resonant grating waveguide structure as platform enhancement for detecting the interaction between fluorescent labeled Boldenone, a non-natural androgenic hormone, and a specific anti-anabolic antibody. We were able to detect a surface coverage of approximately 0.7 ng/mm(2).

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We report a simple methodology to provide complete pulse characterization at the sample plane of a two-photon excited fluorescence (TPEF) microscope. This is achieved by using backward propagating second-harmonic generation (SHG) from starch granules. Without any modification to the microscope, SHG-autocorrelation traces were obtained by using a single starch granule that was placed alongside the biological specimen being imaged. A spectrally resolved SHG autocorrelation was acquired by placing a spectrometer at the output port of the microscope. Complete in situ pulse information is then directly retrieved in an analytical way using the measurement of electric filed by interferometric spectral trace observation (MEFISTO) technique.

Second harmonic generation (SHG) imaging has emerged in recent years as an important laboratory imaging technique since it can provide unique structural information with submicron resolution. It enjoys the benefits of non-invasive interaction establishing this imaging modality as ideal for in vivo investigation of tissue architectures. In this study we present, polarization dependant high resolution SHG images of Caenorhabditis elegans muscles in vivo. We imaged a variety of muscular structures such as body walls, pharynx and vulva. By fitting the experimental data into a cylindrical symmetry spatial model we mapped the corresponding signal distribution of the χ (2) tensor and identified its main axis orientation for different sarcomeres of the earth worm. The cylindrical symmetry was considered to arise from the thick filaments architecture of the inside active volume. Moreover, our theoretical analysis allowed calculating the mean orientation of harmonophores (myosin helical pitch). Ultimately, we recorded and analysed vulvae muscle dynamics, where SHG signal decreased during in vivo contraction.