A method for the determination of the integral refractive index of living cells in suspension by digital holographic microscopy is described. Digital holographic phase contrast images of spherical cells in suspension are recorded, and the radius as well as the integral refractive index are determined by fitting the relation between cell thickness and phase distribution to the measured phase data. The algorithm only requires information about the refractive index of the suspension medium and the image scale of the microscope system. The specific digital holographic microscopy advantage of subsequent focus correction allows a simultaneous investigation of cells in different focus planes. Results obtained from human pancreas and liver tumor cells show that the integral cellular refractive index decreases with increasing cell radius.

We report a mathematically rigorous technique which facilitates the optimization of various optical properties of electromagnetic fields in free space and including scattering interactions. The technique exploits the linearity of electromagnetic fields along with the quadratic nature of the intensity to define specific Optical Eigenmodes (OEi) that are pertinent to the interaction considered. Key applications include the optimization of the size of a focused spot, the transmission through sub-wavelength apertures, and of the optical force acting on microparticles. We verify experimentally the OEi approach by minimising the size of a focused optical field using a superposition of Bessel beams.

We report the focusing of light to generate a subdiffractive, subwavelength focal spot of full width half maximum 222 nm at an operating wavelength of 633 nm using an optical eigenmode approach. Crucially, the spot is created in the focal plane of a microscope objective thus yielding a practical working distance for applications. The optical eigenmode approach is implemented using an optimal superposition of Bessel beams on a spatial light modulator. The effects of partial coherence are also discussed. This far field method is a key advance toward the generation of subdiffractive optical features for imaging and lithographic purposes.

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.

We present an indirect imaging method that measures both amplitude and phase information from a transmissive target. Our method is based on an optical eigenmode decomposition of the light intensity and the first-order cross correlation between a target field and these eigenmodes. We demonstrate that such optical eigenmode imaging does not need any a priori knowledge of the imaging system and corresponds to a compressive full-field sampling, leading to high image extraction efficiencies. Finally, we discuss the implications with respect to second-order correlation imaging.