Øyvind Frette

Testing Fluorescence Lifetime Standards using Two-Photon Excitation and Time-Domain Instrumentation: Rhodamine B, Coumarin 6 and Lucifer Yellow

Having good information about fluorescence lifetime standards is essential for anyone performing lifetime experiments. Using lifetime standards in fluorescence spectroscopy is often regarded as a straightforward process, however, many earlier reports are limited in terms of lifetime concentration dependency, solvents and other technical aspects. We have investigated the suitability of the fluorescent dyes rhodamine B, coumarin 6, and lucifer yellow as lifetime standards, especially to be used with two-photon excitation measurements in the time-domain. We measured absorption and emission spectra for the fluorophores to determine which wavelengths we should use for the excitation and an appropriate detector range. We also measured lifetimes for different concentrations, ranging from 10−2– 10−6 M, in both water, ethanol and methanol solutions. We observed that rhodamine B lifetimes depend strongly on concentration. Coumarin 6 provided the most stable lifetimes, with a negligible dependency on concentration and solvent. Lucifer yellow lifetimes were also found to depend little with concentration. Finally, we found that a mix of two fluorophores (rhodamine B/coumarin 6, rhodamine B/lucifer yellow, and coumarin 6/lucifer yellow) all yielded very similar lifetimes from a double-exponential decay as the separate lifetimes measured from a single-exponential decay. All lifetime measurements were made using two-photon excitation and obtaining lifetime data in the time-domain using time-correlated single-photon counting.

Seasonal variability in inherent optical properties in a western Norwegian fjord

We present measured seasonal variations in the inherent optical properties (the absorption and scattering coefficients) of water in a deep silled fjord (Samnangerfjorden) in western Norway. These were based on measurements taken at monthly intervals during an annual cycle. The measurements also include concentrations of chlorophyll a and yellow substance, which were assumed to dominate the behaviour of the absorption and scattering coefficients. The stations were at three fixed locations, one being placed in the innermost part of the fjord where there is little mixing of fjord water with water from the coastal current. The other two stations were placed at different distances from the mouth of the fjord, so that the water masses are characterized by different amounts of mixing between fjord water and coastal current water. Our data set shows how the absorption and scattering coefficients vary in a Norwegian fjord during an annual cycle, and how they depend on the concentrations of chlorophyll a and yellow substance. Values of the absorption coefficient at 412 nm varied between 0.1 and 2.0 m , and scattering coefficients were also found to vary within this range. Little variation over the spectral range was found for the scattering coefficients, but the absorption coefficient had larger spectral variations. The chlorophyll a concentrations varied from 0.01 to 6.3 mg m , and the concentration of yellow substance, as expressed by its absorption coefficient at 310 nm, was within the range 0.7–7.8 m .

Optical remote sensing of waters with vertical structure

Optical remote sensing of ocean color is a well-established technique that is used to produce maps of marine constituents on a routine basis. Retrieval algorithms used to infer pigment concentrations from measurements of ocean color are usually based on the assumption that the upper ocean column is vertically homogeneous. However, stable stratification of the water column is often encountered in coastal waters and in fjords. This stratification is decisive for the initiation, maintainance, and species composition of phytoplankton blooms. Here we present an optical remote-sensing algorithm with the ability to resolve such a vertical structure of oceanic waters. The vertical structure is assumed to consist of two homogeneous layers with different concentrations of chlorophyll a. The algorithm is designed to determine the chlorophyll-a concentrations of the two layers as well as the thickness of the upper layer. These three parameters influence the ocean color and are simultaneously retrieved through an inverse-modeling technique. This technique consists of using radiative-transfer computations for a coupled atmosphere-ocean system to simulate radiances received in various bands of the satellite sensor and to compare these simulated results with measured radiances. The sum of absolute values of differences between simulated and measured radiances is minimized by use of an optimization algorithm, and the retrieved parameters are those that yield the minimum sum of differences between measured and simulated data. The optimization algorithm that we used in our study is the simulated annealing method, which is an extension of the downhill simplex algorithm. In this study the algorithm was tested on synthetic data generated by the forward model. The results indicate that it should be possible to retrieve vertical variations in the pigment concentration. The synthetic data were generated for spectral bands that coincide with those of the Medium Resolution Imaging Spectrometer sensor, which will be a part of the instrument package of the upcoming Environmental Satellite.

Scattering properties of microalgae: the effect of cell size and cell wall.

The main objective of this work was to investigate how the cell size and the presence of a cell wall influence the scattering properties of the green microalgae Chlamydomonas reinhardtii. The growth cycle of two strains, one with a cell wall and one without, was synchronized to be in the same growth phase. Measurements were conducted at two different phases of the growth cycle on both strains of the algae. It was found that the shape of the scattering phase function was very similar for both strains at both growth phases, but the regular strain with a cell wall scatters more strongly than the wall-less mutant. It was also found that the mutant strain has a stronger increase in scattering than the regular strain, as the algae grow, and that the scattering from the regular strain is more wavelength dependent than from the mutant strain.

Parameterization and analysis of the optical absorption and scattering coefficients in a western Norwegian fjord: a case II water study

Based on statistical analyses of optical properties measured during a whole year of monthly cruises in a Norwegian fjord, we constructed a two-component model for the absorption and scattering coefficients for visible light. The input to the model is the concentrations of yellow substance and chlorophyll a. Because of the presence of a significant amount of nonalgal particles in coastal water, we assume that the absorption and scattering coefficients approach constant background values when the concentration of chlorophyll a approaches zero. The model works reasonably for a variety of optical conditions encountered throughout the year, with a possible exception during a bloom of the Emiliania huxleyi algae in June.

Mueller matrix measurements and modeling pertaining to Spectralon white reflectance standards

The full Mueller matrix for a Spectralon white reflectance standard was measured in the incidence plane, to obtain the polarization state of the scattered light for different angles of illumination. The experimental setup was a Mueller matrix ellipsometer, by which measurements were performed for scattering angles measured relative to the normal of the Spectralon surface from -90° to 90° sampled at every 2.5° for an illumination wavelength of 532 nm. Previously, the polarization of light scattered from Spectralon white reflectance standards was measured only for four of the elements of the Muller matrix. As in previous investigations, the reflection properties of the Spectralon white reflectance standard was found to be close to those of a Lambertian surface for small scattering and illumination angles. At large scattering and illumination angles, all elements of the Mueller matrix were found to deviate from those of a Lambertian surface. A simple empirical model with only two parameters, was developed, and used to simulate the measured results with fairly good accuracy.

Mueller matrix measurements of algae with different shape and size distributions

The full Mueller matrix was measured to obtain the polarization state of the scattered light for a variety of algae with different shapes, wall compositions, sizes, and refractive indices. The experimental setup was a multiple laser Mueller matrix ellipsometer, by which measurements were performed for scattering angles from 16° to 160° sampled at every second degree for wavelengths of 473 nm and 532 nm. Previously, the polarization of light scattered from microalgae was investigated only for a few species, and the Mueller matrix was found to have little variation between the species. In our work a total of 11 algal species were investigated, representing diatoms, dinoflagellates, coccolithophorids, green algae, and a cryptophyte. The selection of species was made to obtain high variability in shape, size, cell wall, and refractive index. As in previous investigations, very small variations were found between species for most of the Mueller matrix elements, but noticeable variations were found for M(11), (M(12)+M(21))/2 and (M(33)+M(44))/2.

Modeling optical properties of human skin using Mie theory for particles with different size distributions and refractive indices

We used size distributions of volume equivalent spherical particles with complex refractive index to model the inherent optical properties (IOPs) in four different layers of human skin at ten different wavelengths in the visible and near-infrared spectral bands. For each layer, we first computed the size-averaged absorption coefficient, scattering coefficient, and asymmetry factor for the collection of particles in a host medium using Mie theory and compared these IOPs in each layer with those obtained from a bio-optical model (BOM). This procedure was repeated, using an optimization scheme, until satisfactory agreement was obtained between the IOPs obtained from the particle size distribution and those given by the BOM. The size distribution as well as the complex refractive index of the particles, obtained from this modeling exercise, can be used to compute the phase matrix, which is an essential input to model polarized light transport in human skin tissue.

Numerical and experimental results for focusing of three-dimensional electromagnetic waves into uniaxial crystals

We present experimental results for focusing of a three-dimensional electromagnetic wave through a plane interface into two different uniaxial crystals, a positive MgF2 crystal and a negative LiNbO3 crystal. These results are compared with numerical results and good agreement is found, both for intensity distributions in various receiving planes and for the locations of the sagittal and tangential focal planes. The theory is briefly outlined both for the exact solution, which includes extraparaxial geometries and double refraction, and for the paraxial solution, in which double refraction is ignored.

Parameterization of the inherent optical properties of Murchison Bay, Lake Victoria

Lake Victoria, Africas largest freshwater lake, suffers greatly from negative changes in biomass of species of fish and also from severe eutrophication. The continuing deterioration of Lake Victorias ecological functions has great long-term consequences for the ecosystem benefits it provides to the countries bordering its shores. However, knowledge about temporal and spatial variations of optical properties and how they relate to lake constituents is important for a number of reasons such as remote sensing, modeling of underwater light fields, and long-term monitoring of lake waters. Based on statistical analysis of data from optical measurements taken during half a year of weekly cruises in Murchison Bay, Lake Victoria, we present a three-component model for the absorption and a two-component model for the scattering of light in the UV and the visible regions of the solar spectrum along with tests of their ranges of validity. The three-component input to the model for absorption is the chlorophyll-a (Chl-a), total suspended materials concentrations, and yellow substance absorption, while the two-component input to the model for scattering is the Chl-a concentration and total suspended materials.