An extended quantitative model for super-resolution optical fluctuation imaging (SOFI)

Super-resolution optical fluctuation imaging (SOFI) provides super-resolution (SR) fluorescence imaging by analyzing fluctuations in the fluorophore emission. The technique has been used both to acquire quantitative SR images and to provide SR biosensing by monitoring changes in fluorophore blinking dynamics. Proper analysis of such data relies on a fully quantitative model of the imaging. However, previous SOFI imaging models made several assumptions that can not be realized in practice. In this work we address these limitations by developing and verifying a fully quantitative model that better approximates real-world imaging conditions. Our model shows that (i) SOFI images are free of bias, or can be made so, if the signal is stationary and fluorophores blink independently, (ii) allows a fully quantitative description of the link between SOFI imaging and probe dynamics, and (iii) paves the way for more advanced SOFI image reconstruction by offering a computationally fast way to calculate SOFI images for arbitrary probe, sample and instrumental properties

Chandra X-Ray Spectroscopy Of The Very Early O Supergiant HD 93129A: Constraints On Wind Shocks And The Mass-Loss Rate

We present an analysis of both the resolved X-ray emission-line profiles and the broad-band X-ray spectrum of the O-2 If* star HD 93129A, measured with the Chandra High Energy Transmission Grating Spectrometer ( HETGS). This star is among the earliest and most massive stars in the Galaxy, and provides a test of the embedded wind-shock scenario in a very dense and powerful wind. A major new result is that continuum absorption by the dense wind is the primary cause of the hardness of the observed X-ray spectrum, while intrinsically hard emission from colliding wind shocks contributes less than 10 per cent of the X-ray flux. We find results consistent with the predictions of numerical simulations of the line-driving instability, including line broadening indicating an onset radius of X-ray emission of several tenths of R-*. Helium-like forbidden-to-intercombination line ratios are consistent with this onset radius, and inconsistent with being formed in a wind-collision interface with the star\u27s closest visual companion at a distance of 100 au. The broad-band X-ray spectrum is fitted with a dominant emission temperature of just kT = 0.6 keV along with significant wind absorption. The broad-band wind absorption and the line profiles provide two independent measurements of the wind mass-loss rate:. M = 5.2(-1.5)(+1.8) x 10(-6) and 6.8(-2.2)(+2.8) x 10(-6) M-circle dot yr(-1), respectively. This is the first consistent modelling of the X-ray line-profile shapes and broad-band X-ray spectral energy distribution in a massive star, and represents a reduction of a factor of 3-4 compared to the standard H alpha mass-loss rate that assumes a smooth wind

Chandra X-ray spectroscopy of the very early O supergiant HD 93129A: constraints on wind shocks and the mass-loss rate

We present analysis of both the resolved X-ray emission line profiles and the broadband X-ray spectrum of the O2 If* star HD 93129A, measured with the Chandra HETGS. This star is among the earliest and most massive stars in the Galaxy, and provides a test of the embedded wind shock scenario in a very dense and powerful wind. A major new result is that continuum absorption by the dense wind is the primary cause of the hardness of the observed X-ray spectrum, while intrinsically hard emission from colliding wind shocks contributes less than 10% of the X-ray flux. We find results consistent with the predictions of numerical simulations of the line-driving instability, including line broadening indicating an onset radius of X-ray emission of several tenths Rstar. Helium-like forbidden-to-intercombination line ratios are consistent with this onset radius, and inconsistent with being formed in a wind-collision interface with the star's closest visual companion at a distance of ~100 AU. The broadband X-ray spectrum is fit with a dominant emission temperature of just kT = 0.6 keV along with significant wind absorption. The broadband wind absorption and the line profiles provide two independent measurements of the wind mass-loss rate: Mdot = 5.2_{-1.5}^{+1.8} \times 10^{-6} Msun/yr and Mdot = 6.8_{-2.2}^{+2.8} \times 10^{-6} Msun/yr, respectively. This is the first consistent modeling of the X-ray line profile shapes and broadband X-ray spectral energy distribution in a massive star, and represents a reduction of a factor of 3 to 4 compared to the standard H-alpha mass-loss rate that assumes a smooth wind.Comment: Accepted for publication in Monthly Notices of the Royal Astronomical Society. 12 pages, 10 figures (incl. 5 color

Infection of Mice with the Agent of Human Granulocytic Ehrlichiosis after Different Routes of Inoculation

Population kinetics of the agent of human granulocytic ehrlichiosis (aoHGE) were examined after needle and tickborne inoculation of C3H mice. Blood, skin, lung, spleen, liver, kidney, brain, lymph node, and bone marrow samples were analyzed by using real-time polymerase chain reaction (PCR) at various intervals after inoculation, using a p44 gene target. The highest number of copies of the p44 gene target occurred in blood and bone marrow samples, emphasizing aoHGE leukocytotropism. Numbers of copies of the p44 gene target in other tissues reflected vascular perfusion rather than replication. Needle-inoculated infected mice had earlier dissemination, but kinetics of infection in both groups were parallel, with declining rates of infection by day 20 and recovery in some mice on days 20-60 after inoculation. On the basis of an aoHGE lysate ELISA, mice seroconverted by day 10 after inoculation. Therefore, real-time PCR is useful for quantitative studies with the aoHGE in experimental infections, and results showed that needle inoculation can be used to study the aoHGE infection because of its similarity to tickborne inoculatio

Photoactivatable Fluorophore for Stimulated Emission Depletion (STED) Microscopy and Bioconjugation Technique for Hydrophobic Labels

The use of photoactivatable dyes in STED microscopy has so far been limited by two—photon activation through the STED beam and by the fact that photoactivatable dyes are poorly solvable in water. Here we report ONB‐2SiR, a fluorophore that can be both photoactivated in the UV and specifically de‐excited by STED at 775 nm. Likewise, we introduce a conjugation and purification protocol to effectively label primary and secondary antibodies with moderately water‐soluble dyes. Greatly reducing dye aggregation, our technique provides a defined and tunable degree of labeling, and improves the imaging performance of dye conjugates in general

Measuring Mass-Loss Rates And Constraining Shock Physics Using X-Ray Line Profiles Of O Stars From The Chandra Archive

We quantitatively investigate the extent of wind absorption signatures in the X-ray grating spectra of all non-magnetic, effectively single O stars in the Chandra archive via line profile fitting. Under the usual assumption of a spherically symmetric wind with embedded shocks, we confirm previous claims that some objects show little or no wind absorption. However, many other objects do show asymmetric and blueshifted line profiles, indicative of wind absorption. For these stars, we are able to derive wind mass-loss rates from the ensemble of line profiles, and find values lower by an average factor of 3 than those predicted by current theoretical models, and consistent with Hα if clumping factors of fcl ≈ 20 are assumed. The same profile fitting indicates an onset radius of X-rays typically at r ≈ 1.5R*, and terminal velocities for the X-ray emitting wind component that are consistent with that of the bulk wind. We explore the likelihood that the stars in the sample that do not show significant wind absorption signatures in their line profiles have at least some X-ray emission that arises from colliding wind shocks with a close binary companion. The one clear exception is ζ Oph, a weak-wind star that appears to simply have a very low mass-loss rate. We also reanalyse the results from the canonical O supergiant ζ Pup, using a solar-metallicity wind opacity model and find M˙=1.8×10−6 M⊙yr−1, consistent with recent multiwavelength determinations

A Coordinated X-ray and Optical Campaign on the Nearest Massive Eclipsing Binary, Delta Ori Aa: I. Overview of the X-ray Spectrum

We present an overview of four phase-constrained Chandra HETGS X-ray observations of Delta Ori A. Delta Ori A is actually a triple system which includes the nearest massive eclipsing spectroscopic binary, Delta Ori Aa, the only such object which can be observed with little phase-smearing with the Chandra gratings. Since the fainter star, Delta Ori Aa2, has a much lower X-ray luminosity than the brighter primary, Delta Ori A provides a unique system with which to test the spatial distribution of the X-ray emitting gas around Delta Ori Aa1 via occultation by the photosphere of and wind cavity around the X-ray dark secondary. Here we discuss the X-ray spectrum and X-ray line profiles for the combined observation, having an exposure time of nearly 500 ksec and covering nearly the entire binary orbit. Companion papers discuss the X-ray variability seen in the Chandra spectra, present new space-based photometry and ground-based radial velocities simultaneous with the X-ray data to better constrain the system parameters, and model the effects of X-rays on the optical and UV spectrum. We find that the X-ray emission is dominated by embedded wind shock emission from star Aa1, with little contribution from the tertiary star Ab or the shocked gas produced by the collision of the wind of Aa1 against the surface of Aa2. We find a similar temperature distribution to previous X-ray spectrum analyses. We also show that the line half-widths are about $0.3-0.5\times$ the terminal velocity of the wind of star Aa1. We find a strong anti-correlation between line widths and the line excitation energy, which suggests that longer-wavelength, lower-temperature lines form farther out in the wind. Our analysis also indicates that the ratio of the intensities of the strong and weak lines of \ion{Fe}{17} and \ion{Ne}{10} are inconsistent with model predictions, which may be an effect of resonance scatteringComment: accepted by ApJ; revised according to ApJ proo

Accelerator experiments with soft protons and hyper-velocity dust particles: application to ongoing projects of future X-ray missions

We report on our activities, currently in progress, aimed at performing accelerator experiments with soft protons and hyper-velocity dust particles. They include tests of different types of X-ray detectors and related components (such as filters) and measurements of scattering of soft protons and hyper-velocity dust particles off X-ray mirror shells. These activities have been identified as a goal in the context of a number of ongoing space projects in order to assess the risk posed by environmental radiation and dust and qualify the adopted instrumentation with respect to possible damage or performance degradation. In this paper we focus on tests for the Silicon Drift Detectors (SDDs) used aboard the LOFT space mission. We use the Van de Graaff accelerators at the University of T\"ubingen and at the Max Planck Institute for Nuclear Physics (MPIK) in Heidelberg, for soft proton and hyper-velocity dust tests respectively. We present the experimental set-up adopted to perform the tests, status of the activities and some very preliminary results achieved at present time.Comment: Proceedings of SPIE, Vol. 8443, Paper No. 8443-24, 201