Parallel single cell analysis on an integrated microfluidic platform for cell trapping, lysis and analysis

We report here a novel and easily scalable microfluidic platform for the parallel analysis of hundreds of individual cells, with controlled single cell trapping, followed by their lysis and subsequent retrieval of the cellular content for on-chip analysis. The device consists of a main channel and an array of shallow side channels connected to the main channel via trapping structures. Cells are individually captured in dam structures by application of a negative pressure from an outlet reservoir, lyzed on site and the cellular content controllably extracted and transported in the individual side channels for on-chip analysis.\u

Chandra Detection of X-ray Absorption Associated with a Damped Lyman Alpha System

We have observed three quasars, PKS 1127-145, Q 1331+171 and Q0054+144, with the ACIS-S aboard the Chandra X-ray Observatory, in order to measure soft X-ray absorption associated with intervening 21-cm and damped Ly$\alpha$ absorbers. For PKS 1127-145, we detect absorption which, if associated with an intervening z_{abs}=0.312 absorber, implies a metallicity of 23% solar. If the absorption is not at z_{abs}=0.312, then the metallicity is still constrained to be less than 23% solar. The advantage of the X-ray measurement is that the derived metallicity is insensitive to ionization, inclusion of an atom in a molecule, or depletion onto grains. The X-ray absorption is mostly due to oxygen, and is consistent with the oxygen abundance of 30% solar derived from optical nebular emission lines in a foreground galaxy at the redshift of the absorber. For Q1331+171 and Q 0054+144, only upper limits were obtained, although the exposure times were intentionally short, since for these two objects we were interested primarily in measuring flux levels to plan for future observations. The imaging results are presented in a companion paper.Comment: 23 pages, 6 figures, accepted for publication in the Astrophysical Journa

Memory CD4 T cell subsets are kinetically heterogeneous and replenished from naive T cells at high levels

Characterising the longevity of immunological memory requires establishing the rules underlying the renewal and death of peripheral T cells. However, we lack knowledge of the population structure and how self-renewal and de novo influx contribute to maintenance of memory compartments. Here, we characterise the kinetics and structure of murine CD4 T cell memory subsets by measuring the rates of influx of new cells and using detailed timecourses of DNA labelling that also distinguish the behaviour of recently divided and quiescent cells. We find that both effector and central memory CD4 T cells comprise subpopulations with highly divergent rates of turnover, and show that inflows of new cells sourced from the naive pool strongly impact estimates of memory cell lifetimes and division rates. We also demonstrate that the maintenance of CD4 T cell memory subsets in healthy mice is unexpectedly and strikingly reliant on this replenishment

Towards simultaneous electrical and optical investigation of BLMS using a novel microfluidic device

We firstly describe the influence of the phospholipid (PL) composition of bilayer lipid membrane on their electrical properties: (i) the more unsaturations in the tail, the earlier the BLM breakdown and (ii) the bulkier the head group, the less stable the membrane. Secondly, we design and fabricate novel devices that couple such electri-cal characterization to optical investigation and that enable the preparation of asym-metrical membranes: a “macro” device including a drilled PMMA plate as well as microfluidic device consisting of a glass-teflon foil-glass sandwich

Microstamped petri dishes for scanning electrochemical microscopy analysis of arrays of microtissues

While scanning electrochemical microscopy (SECM) is a powerful technique for non-invasive analysis of cells, SECM-based assays remain scarce and have been mainly limited so far to single cells, which is mostly due to the absence of suitable platform for experimentation on 3D cellular aggregates or microtissues. Here, we report stamping of a Petri dish with a microwell array for large-scale production of microtissues followed by their in situ analysis using SECM. The platform is realized by hot embossing arrays of microwells (200 mum depth; 400 mum diameter) in commercially available Petri dishes, using a PDMS stamp. Microtissues form spontaneously in the microwells, which is demonstrated here using various cell lines (e.g., HeLa, C2C12, HepG2 and MCF-7). Next, the respiratory activity of live HeLa microtissues is assessed by monitoring the oxygen reduction current in constant height mode and at various distances above the platform surface. Typically, at a 40 mum distance from the microtissue, a 30% decrease in the oxygen reduction current is measured, while above 250 mum, no influence of the presence of the microtissues is detected. After exposure to a model drug (50% ethanol), no such changes in oxygen concentration are found at any height in solution, which reflects that microtissues are not viable anymore. This is furthermore confirmed using conventional live/dead fluorescent stains. This live/dead assay demonstrates the capability of the proposed approach combining SECM and microtissue arrays formed in a stamped Petri dish for conducting cellular assays in a non-invasive way on 3D cellular models

Effect of antioxidant supplementation on the total yield, oxidative stress levels and multipotency of bone marrow-derived human mesenchymal stromal cells

Bone marrow-derived multipotent mesenchymal stromal cells (MSCs) are the most frequently investigated cell type for potential regenerative strategies because they are relatively easy to isolate and are able to differentiate into several mesenchymal lineages. Unfortunately, during ex vivo culture, MSCs present gradual loss of differentiation potential and reduced clinical efficacy. Reactive oxygen species (ROS) are associated with oxidative damage and accumulate during MSC expansion. Because ROS are believed to be involved in the loss of multipotency, we hypothesized that compounds with antioxidant activity have the capacity to scavenge ROS, prevent cellular damage, and rescue culture-induced loss of multipotency. In this manuscript, we show that antioxidant supplementation can partially rescue the loss of alkaline phosphatase expression induced by oxidizing agents and increases the yield of hMSCs, when supplemented to a fresh bone marrow aspirate. Concomitantly, oxidative DNA damage and ROS levels in hMSCs were reduced by antioxidants. We conclude that antioxidant supplementation during MSC expansion reduces the DNA damage load and increases the MSC yield

Novel Electroweak Symmetry Breaking Conditions From Quantum Effects In The MSSM

We present, in the context of the Minimal Supersymmetric Standard Model, a detailed one-loop analytic study of the minimization conditions of the effective potential in the Higgs sector. Special emphasis is put on the role played by $Str M^4$ in the determination of the electroweak symmetry breaking conditions, where first and second order derivatives of the effective potential are systematically taken into account. Novel, necessary (and sufficient in the Higgs sector) model-independent constraints, are thus obtained analytically, leading to new theoretical lower and upper bounds on $\tan \beta$. Although fully model-independent, these bounds are found to be much more restrictive than the existing model-dependent ones! A first illustration is given in the context of a SUGRA-GUT motivated scenario.Comment: Latex, 45 pages, 5 figure

Spitzer Mapping of PAHs and H2 in Photodissociation Regions

The mid-infrared (MIR) spectra of dense photodissociation regions (PDRs) are typically dominated by emission from polycyclic aromatic hydrocarbons (PAHs) and the lowest pure rotational states of molecular hydrogen (H2); two species which are probes of the physical properties of gas and dust in intense UV radiation fields. We utilize the high angular resolution of the Infrared Spectrograph on the Spitzer Space Telescope to construct spectral maps of the PAH and H2 features for three of the best studied PDRs in the galaxy, NGC 7023, NGC 2023 and IC 63. We present spatially resolved maps of the physical properties, including the H2 ortho-to-para ratio, temperature, and G_o/n_H. We also present evidence for PAH dehydrogenation, which may support theories of H2 formation on PAH surfaces, and a detection of preferential self-shielding of ortho-H2. All PDRs studied exhibit average temperatures of ~500 - 800K, warm H2 column densities of ~10^20 cm^-2, G_o/n_H ~ 0.1 - 0.8, and ortho-to-para ratios of ~ 1.8. We find that while the average of each of these properties is consistent with previous single value measurements of these PDRs, when available, the addition of spatial resolution yields a diversity of values with gas temperatures as high as 1500 K, column densities spanning ~ 2 orders of magnitude, and extreme ortho-to-para ratios of 3.Comment: 14 figure

Emission of photon echoes in a strongly scattering medium

We observe the two- and three-pulse photon echo emission from a scattering powder, obtained by grinding a Pr$^{3+}$:Y$_2$SiO$_5$ rare earth doped single crystal. We show that the collective emission is coherently constructed over several grains. A well defined atomic coherence can therefore be created between randomly placed particles. Observation of photon echo on powders as opposed to bulk materials opens the way to faster material development. More generally, time-domain resonant four-wave mixing offers an attractive approach to investigate coherent propagation in scattering media

Tests with a Carlina-type diluted telescope; Primary coherencing

Studies are under way to propose a new generation of post-VLTI interferometers. The Carlina concept studied at the Haute- Provence Observatory is one of the proposed solutions. It consists in an optical interferometer configured like a diluted version of the Arecibo radio telescope: above the diluted primary mirror made of fixed cospherical segments, a helium balloon (or cables suspended between two mountains), carries a gondola containing the focal optics. Since 2003, we have been building a technical demonstrator of this diluted telescope. First fringes were obtained in May 2004 with two closely-spaced primary segments and a CCD on the focal gondola. We have been testing the whole optical train with three primary mirrors. The main aim of this article is to describe the metrology that we have conceived, and tested under the helium balloon to align the primary mirrors separate by 5-10 m on the ground with an accuracy of a few microns. The servo loop stabilizes the mirror of metrology under the helium balloon with an accuracy better than 5 mm while it moves horizontally by 30 cm in open loop by 10-20 km/h of wind. We have obtained the white fringes of metrology; i.e., the three mirrors are aligned (cospherized) with an accuracy of {\approx} 1 micron. We show data proving the stability of fringes over 15 minutes, therefore providing evidence that the mechanical parts are stabilized within a few microns. This is an important step that demonstrates the feasibility of building a diluted telescope using cables strained between cliffs or under a balloon. Carlina, like the MMT or LBT, could be one of the first members of a new class of telescopes named diluted telescopes.Comment: 18 pages, 17 figures, A&A, accepte