Signal analysis of total internal reflection fluorescent speckle microscopy (TIR-FSM) and wide-field epi-fluorescence FSM of the actin cytoskeleton and focal adhesions in living cells

Fluorescent speckle microscopy (FSM) uses low levels of fluorescent proteins to create fluorescent speckles on cytoskeletal polymers in high‐resolution fluorescence images of living cells. The dynamics of speckles over time encode subunit turnover and motion of the cytoskeletal polymers. We sought to improve on current FSM technology by first expanding it to study the dynamics of a non‐polymeric macromolecular assembly, using focal adhesions as a test case, and second, to exploit for FSM the high contrast afforded by total internal reflection fluorescence microscopy (TIR‐FM). Here, we first demonstrate that low levels of expression of a green fluorescent protein (GFP) conjugate of the focal adhesion protein, vinculin, results in clusters of fluorescent vinculin speckles on the ventral cell surface, which by immunofluorescence labelling of total vinculin correspond to sparse labelling of dense focal adhesion structures. This demonstrates that the FSM principle can be applied to study focal adhesions. We then use both GFP‐vinculin expression and microinjected fluorescently labelled purified actin to compare quantitatively the speckle signal in FSM images of focal adhesions and the actin cytoskeleton in living cells by TIR‐FM and wide‐field epifluorescence microscopy. We use quantitative FSM image analysis software to define two new parameters for analysing FSM signal features that we can extract automatically: speckle modulation and speckle detectability. Our analysis shows that TIR‐FSM affords major improvements in these parameters compared with wide‐field epifluorescence FSM. Finally, we find that use of a crippled eukaryotic expression promoter for driving low‐level GFP‐fusion protein expression is a useful tool for FSM imaging. When used in time‐lapse mode, TIR‐FSM of actin and GFP‐conjugated focal adhesion proteins will allow quantification of molecular dynamics within interesting macromolecular assemblies at the ventral surface of living cells.     

Forgotten pioneers of color order. Part I: Gaspard Grégoire (1751–1846)

Some 100 years before Albert Munsell developed his color order system, French silk merchant and inventor of a technology for producing works of art in silk velours, Gaspard Grégoire, introduced a color order system based on the color attributes hue, (relative) chroma, and lightness. Conceived in the mid‐1780s, an atlas with 1350 samples was produced before 1813 and found use in French Royal manufacturing operations and educational institutions. It was followed a few years later by one with 343 samples. Grégoire's work was subsequently overshadowed by Michel‐Eugene Chevreul's more complicated and less intuitive hemispherical system of 1839. © 2007 Wiley Periodicals, Inc. Col Res Appl, 33, 5–9, 2008     

From Long Infrared to Very Long Infrared Wavelength Focal Plane Arrays Made with HgCdTe n + n ?/ p Ion Implantation Technology

This paper aims at studying the feasibility of very long infrared wavelength (VLWIR) (12–18 μm) focal plane arrays using n-on-p planar ion-implanted technology. To explore and analyze the feasibility of such VLWIR detectors, a set of four Cd _x Hg_1−x Te LPE layers with an 18 μ cutoff at 50 K has been processed at Defir (LETI/LIR–Sofradir joint laboratory), using both our “standard” n-on-p process and our improved low dark current process. Several 320 × 256 arrays, 30-μm pitch, have been hybridized on standard Sofradir readout circuits and tested. Small dimension test arrays characterization is also presented. Measured photonic currents with a 20°C black body suggest an internal quantum efficiency above 50%. Typical I(V) curves and thermal evolution of the saturation current are discussed, showing that standard photodiodes remain diffusion limited at low biases for temperatures down to 30 K. Moreover, the dark current gain brought by the improved process is clearly visible for temperatures higher than 40 K. Noise measurements are also discussed showing that a very large majority of detectors appeared background limited under usual illumination and biases. In our opinion, such results demonstrate the feasibility of high-performance complex focal plane arrays in the VLWIR range at medium term.     

Experimental determination of NH<Subscript>4</Subscript>NO<Subscript>3</Subscript>-KNO<Subscript>3</Subscript> binary phase diagram

The solid-state phase transitions in ammonium nitrate (AN)-potassium nitrate (KN) system, and the equilibrium AN-KN phase diagram have been determined by using differential scanning calorimetry and high-temperature in situ x-ray diffractometry. Sample preparation was performed in a special “dry room” with very low humidity. A single phase region (AN III) with no phase transitions to 373 K was observed in the composition range 5 to 20% KN; this is critical for use in air bag gas generators. The high-temperature KN phase (KN I) has a wide range of stability from 20 to 100 wt.% KN. There are one eutectic, two eutectoid, three peritectoid, and one congruent transformations in this phase diagram. Two new nonstoichiometric phases were found at lower temperatures in the mid-composition range between the AN and KN terminal solid solutions. Details of the phase equilibria are presented.     

Microstructures formed by spray freezing of food fats

The spray-freezing of two food fats, tripalmitin (PPP) and cocoa butter (CB) and mixtures thereof, has been modeled experimentally using a novel single droplet freezing apparatus configured so that temperature profiles or samples for microstructure analysis can be obtained. For 2 mm diameter droplets suspended in a cold air flow at temperatures around 2–15°C, initial cooling rates were on the order of 10 K s⁻¹ and the temperature profiles could be correlated directly to DSC data collected at 20 K min⁻¹, indicating that minimal supercooling of the materials occurred in the droplet form. Microstructure analysis confirmed that PPP crystallized preferentially in mixtures, and that the surface structure was very sensitive to storage conditions. The bulk structure was much less sensitive, and the internal microstructure of the PPP droplets revealed distinct nucleation sites, which were absent from the CB: These persisted in the mixtures up to 50 wt%. X-ray analysis indicated that the fats crystallized in their more stable forms, namely, β for PPP and Form V/V1 in CB.     

Quantum Dots-Based Optical Fiber Temperature Sensors Fabricated by Layer-by-Layer

Two types of CdTe quantum dots of different sizes (4 and 5 nm) were successfully deposited on optical fibers using the layer-by-layer electrostatic self-assembly method. The sensors showed a linear and reversible variation of the emission wavelength for a temperature range from 30 degC to 100 degC, with a sensitivity of 0.2 nm/degC     

Formation mechanism of 21R AlN-polytypoids in aluminothermic reduction and nitridation process

Synthesis of 21R AlN polytypoids was investigated using Al and ultrafine SiO₂ powder in a flowing nitrogen atmosphere by means of thermal gravity (TG), differential scanning calorimeter (DSC), X-ray diffractometer (XRD) and scanning electronic microscope (SEM) linked with energy dispersive spectrometer (EDS). The results showed that the formation mechanism of AlN polytypoids was different from that in reaction sintering process using Si₃N₄, AlN, Al₂O₃ and other sintering additive as raw materials. It was suggested that firstly Al reduce SiO₂ into Si and is also nitrided into AlN, then AlN, Al₂O₃ and SiO₂ dissolve into silicon liquid until the AlN polytypoids precipitate in saturated liquid in a flowing nitrogen atmosphere at lower than 1700 °C.     

Sensitivity analysis in conjunction with evidence theory representations of epistemic uncertainty

Three applications of sampling-based sensitivity analysis in conjunction with evidence theory representations for epistemic uncertainty in model inputs are described: (i) an initial exploratory analysis to assess model behavior and provide insights for additional analysis; (ii) a stepwise analysis showing the incremental effects of uncertain variables on complementary cumulative belief functions and complementary cumulative plausibility functions; and (iii) a summary analysis showing a spectrum of variance-based sensitivity analysis results that derive from probability spaces that are consistent with the evidence space under consideration.