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AVP and atrial natriuretic peptide (ANP) have opposite effects in the kidney. AVP induces antidiuresis by insertion of aquaporin-2 (AQP2) water channels into the plasma membrane of collecting duct principal cells. ANP acts as a diuretic factor. An ANP- and nitric oxide (NO)/soluble guanylate cyclase (sGC)-induced insertion of AQP2 into the plasma membrane is reported from different models. However, functional data on the insertion of AQP2 is missing. We used primary cultured inner medullary collecting duct (IMCD) cells and digital holographic microscopy, calcein-quenching measurements, and immunofluorescence and Western blotting to analyze the effects of ANP and NO donors on AQP2 phosphorylation, membrane expression, and water permeability. While AVP led to acceleration in osmotically induced swelling, ANP had no effect. However, in AVP-pretreated cells ANP significantly decreased the kinetics of cell swelling. This effect was mimicked by 8-bromo-cGMP and blunted by PKG inhibition. Stimulation of the NO/sGC pathway or direct activation of sGC with BAY 58-2667 had similar effects to ANP. In cells treated with AVP, AQP2 was predominantly localized in the plasma membrane, and after additional incubation with ANP AQP2 was mostly localized in the cytosol, indicating an increased retrieval of AQP2 from the plasma membrane by ANP. Western blot analysis showed that ANP was able to reduce AVP-induced phosphorylation of AQP2 at position S256. In conclusion, we show that the diuretic action of ANP or NO in the IMCD involves a decreased localization of AQP2 in the plasma membrane which is mediated by cGMP and PKG. AQP2; cell volume kinetics; cGMP-generating signaling pathways; natriuretic peptides; water homeostasis; uroguanylin, phosphodiesterase-5

In temporal phase-shifting-based digital holographic microscopy, high-resolution phase contrast imaging requires optimized conditions for hologram recording and phase retrieval. To optimize the phase resolution, for the example of a variable three-step algorithm, a theoretical analysis on statistical errors, digitalization errors, uncorrelated errors, and errors due to a misaligned temporal phase shift is carried out. In a second step the theoretically predicted results are compared to the measured phase noise obtained from comparative experimental investigations with several coherent and partially coherent light sources. Finally, the applicability for noise reduction is demonstrated by quantitative phase contrast imaging of pancreas tumor cells. OCIS codes: 090.0090, 090.1995, 100.5070, 180.0180.

Digital holography enables a multifocus quantitative phase microscopy for the investigation of reflective surfaces and for marker-free live cell imaging. For digital holographic long-term investigations of living cells an automated (subsequent) robust and reliable numerical focus adjustment is of particular importance. Four numerical methods for the determination of the optimal focus position in the numerical reconstruction and propagation of the complex object waves of pure phase objects are characterized, compared, and adapted to the requirements of digital holographic microscopy. Results from investigations of an engineered surface and human pancreas tumor cells demonstrate the applicability of Fourier-weighting- and gradient-operator-based methods for robust and reliable automated subsequent numerical digital holographic focusing. OCIS codes: 090.1995, 180.0180.

Digital holographic microscopy enables a quantitative phase contrast metrology that is suitable for the investigation of reflective surfaces as well as for the marker-free analysis of living cells. The digital holographic feature of (subsequent) numerical focus adjustment makes possible applications for multifocus imaging. An overview of digital holographic microscopy methods is described. Applications of digital holographic microscopy are demonstrated by results obtained from livings cells and engineered surfaces. OCIS codes: 090.0090, 180.6900.

A parameter-optimized off-axis setup for digital holographic microscopy is presented for simultaneous, high-resolution, full-field quantitative amplitude and quantitative phase-contrast microscopy and the detection of changes in optical path length in transparent objects, such as undyed living cells. Numerical reconstruction with the described nondiffractive reconstruction method, which suppresses the zero order and the twin image, requires a mathematical model of the phase-difference distribution between the object wave and the reference wave in the hologram plane. Therefore an automated algorithm is explained that determines the parameters of the mathematical model by carrying out the discrete Fresnel transform. Furthermore the relationship between the axial position of the object and the reconstruction distance, which is required for optimization of the lateral resolution of the holographic images, is derived. The lateral and the axial resolutions of the system are discussed and quantified by application to technical objects and to living cells. OCIS codes: 090.1760, 120.3180, 170.1530, 180.0180.

A two-wavelength method for endoscopic topography reconstruction is introduced that can be applied to out-of-plane sensitive electronic-speckle-pattern interferometry systems based on rigid endoscope imaging systems. The surface measurement is performed by detection of the phase-difference distribution affected by a change in the applied laser wavelength. Furthermore, the off-axis endoscopic illumination geometry is taken into account by an approximation. Experimental results of the characterization of the endoscopic surface reconstruction technique and the measurement accuracy obtained are described and discussed. Finally, the applicability of the method is demonstrated with results from the topographic reconstruction of a free-form surface. [c] 2004 Optical Society of America OCIS codes: 120.6160, 120.2880.

Purpose: The conventional impression technique for manufacturing facial prostheses has the disadvantage of deforming the soft tissues because of the tension caused by the impression material, as well as causing discomfort to the patient. The purpose of this study was to establish a system that allows contact-free reproduction of the facial surface combined with computer-assisted design and fabrication of facial prostheses. Materials and Methods: Three-dimensional data of the facial surface were obtained using an optical acquisition system based on the method of phase-measuring profilometry. A sensor head with a fringe projector and two CCD cameras for photogrammetric triangulation were used in connection with a PC for measurement control and data evaluation. Software for computer-assisted design of the facial surface to be reconstructed was developed. A prototype facial prosthesis was fabricated using stereolithography. The system was tested using a modified puppet head. First clinical tests were performed with a patient who had undergone maxillofacial surgery including the resection of one eye. Results: Three dimensional data acquisition and imaging allow visualization of a whole face without causing tension or neuromuscular reaction. As surface brightness is also part of the digital model, it is even more realistic than a plaster cast. The stereolithographic object showed good marginal fit and satisfactory shape. Conclusion: The presented technique allows three-dimensional data reproduction of the facial surface, computer-assisted design of a facial prosthesis, and transfer to a rapid prototyping unit. The system has obvious advantages over conventional impression techniques. Further clinical trials are planned to evaluate the clinical success of the technique. [ABSTRACT FROM AUTHOR]

An endoscope electronic-speckle-pattern interferometer (ESPI) camera system is presented that can be applied to examinations of technical objects as well as for in vitro and in vivo minimal invasive medical diagnostics. Integration of optical fibers for the guidance of a cw-laser beam and an endoscopic imaging system yield a compact ESPI system that opens up new possibilities for highly sensitive interferometric intracavity inspection under handheld conditions. A CCD camera in combination with a fast frame-grabber system allows dynamic image subtractions at a frequency rate of as much as 25 Hz with high fringe contrast. Results from investigations of technical objects and biological objects in vitro and in vivo are obtained. In endoscopic minimal invasive therapy this method could substitute for the missing operator's tactile contact with the treated tissue by replacing it with visual information (endoscopic taction). [C] 2000 Optical Society of America OCIS codes: 120.3180, 120.4290.