In situ fiber-optical monitoring of cytosolic calcium in tissue explant cultures

We present a fluorescence-lifetime based method for monitoring cell and tissue activity in situ, during cell culturing and in the presence of a strong autofluorescence background. The miniature fiber-optic probes are easily incorporated in the tight space of a cell culture chamber or in an endoscope. As a first application we monitored the cytosolic calcium levels in porcine tracheal explant cultures using the Calcium Green-5N (CG5N) indicator. Despite the simplicity of the optical setup we are able to detect changes of calcium concentration as small as 2.5 nM, with a monitoring time resolution of less than 1 s.Comment: 26 pages, 6 figures Article first published online: 11 SEP 2013. appears in J. Biophoton. (2013

Characterization of optical properties of ZnO nanoparticles for quantitative imaging of transdermal transport

Widespread applications of ZnO nanoparticles (NP) in sun-blocking cosmetic products have raised safety concerns related to their potential transdermal penetration and resultant cytotoxicity. Nonlinear optical microscopy provides means for high-contrast imaging of ZnO NPs lending in vitro and in vivo assessment of the nanoparticle uptake in skin, provided their nonlinear optical properties are characterized. We report on this characterization using ZnO NP commercial product, Zinclear, mean-sized 21 nm. Two-photon action cross-section of this bandgap material (Ebg = 3.37 eV, λbg = 370 nm) measured by two techniques yielded consistent results of ηZnOσZnO(2ph) = 6.2 ± 0.8 μGM at 795 nm, and 32 ± 6 μGM at 770 nm per unit ZnO crystal cell, with the quantum efficiency of ηZnO = (0.9 ± 0.2) %. In order to demonstrate the quantitative imaging, nonlinear optical microscopy images of the excised human skin topically treated with Zinclear were acquired and processed using σZnO(2ph) and ηZnOvalues yielding nanoparticle concentration map in skin. Accumulations of Zinclear ZnO nanoparticles were detected only on the skin surface and in skin folds reaching concentrations of 800 NPs per μm3

Silicon avalanche photodiodes as detectors for photon correlation experiments

In view of time correlated photon-counting experiments using wavelengths at the red end of the electromagnetic spectrum, we developed a simple electronic circuit for periodical gated quenching of silicon avalanche photodiodes. We compare the performance of this device with commercially available passive and active quenching modules and a reference photomultiplier. The detection system's nonlinearities, i.e., dead time and afterpulsing, lead to direct and indirect distortions of photocount correlation functions. We characterize this nonlinear behavior by measuring intensity auto-and cross-correlation functions and supply nonlinearity parameters for each of the four detection systems. In addition, transfer functions are given which allow an estimate for the highest count rates accessible for each detection system