Depth profiling of gold nanoparticles and characterization of point spread functions in reconstructed and human skin using multiphoton microscopy |
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Authors: | Hagar I Labouta Martina Hampel Sibylle Thude Katharina Reutlinger Karl‐Heinz Kostka Marc Schneider |
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Institution: | 1. Department of Pharmaceutical Nanotechnology, Saarland University, Campus A4 1, 66123 Saarbrücken, Germany;2. Department of Pharmaceutics, Faculty of Pharmacy, University of Alexandria, Alexandria, Egypt;3. Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany;4. Department of Plastic and Hand Surgery, Caritaskrankenhaus, 66822 Lebach, Germany |
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Abstract: | Multiphoton microscopy has become popular in studying dermal nanoparticle penetration. This necessitates studying the imaging parameters of multiphoton microscopy in skin as an imaging medium, in terms of achievable detection depths and the resolution limit. This would simulate real‐case scenarios rather than depending on theoretical values determined under ideal conditions. This study has focused on depth profiling of sub‐resolution gold nanoparticles (AuNP) in reconstructed (fixed and unfixed) and human skin using multiphoton microscopy. Point spread functions (PSF) were determined for the used water‐immersion objective of 63×/NA = 1.2. Factors such as skin‐tissue compactness and the presence of wrinkles were found to deteriorate the accuracy of depth profiling. A broad range of AuNP detectable depths (20–100 μm) in reconstructed skin was observed. AuNP could only be detected up to ~14 μm depth in human skin. Lateral (0.5 ± 0.1 μm) and axial (1.0 ± 0.3 μm) PSF in reconstructed and human specimens were determined. Skin cells and intercellular components didn't degrade the PSF with depth. In summary, the imaging parameters of multiphoton microscopy in skin and practical limitations encountered in tracking nanoparticle penetration using this approach were investigated. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim) |
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Keywords: | multiphoton imaging gold nanoparticles skin nanoparticle penetration point spread function nanoparticle depth profiling |
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