Two-photon transitions driven by a combination of diode and femtosecond lasers

We report on the combined action of a cw diode laser and a train of ultrashort pulses when each of them drives one step of the 5S-5P-5D two-photon transition in rubidium vapor. The fluorescence from the 6P_{3/2} state is detected for a fixed repetition rate of the femtosecond laser while the cw-laser frequency is scanned over the rubidium D_{2} lines. This scheme allows for a velocity selective spectroscopy in a large spectral range including the 5D_{3/2} and 5D_{5/2} states. The results are well described in a simplified frequency domain picture, considering the interaction of each velocity group with the cw laser and a single mode of the frequency comb.Comment: 4 pages, 4 figure

KATP Channel Openers Have Opposite Effects on Mitochondrial Respiration Under Different Energetic Conditions

Mitochondrial (m) KATP channel opening has been implicated in triggering cardiac preconditioning. Its consequence on mitochondrial respiration, however, remains unclear. We investigated the effects of two different KATP channel openers and antagonists on mitochondrial respiration under two different energetic conditions. Oxygen consumption was measured for complex I (pyruvate/malate) or complex II (succinate with rotenone) substrates in mitochondria from fresh guinea pig hearts. One of two mKATP channel openers, pinacidil or diazoxide, was given before adenosine diphosphate in the absence or presence of an mKATP channel antagonist, glibenclamide or 5-hydroxydecanoate. Without ATP synthase inhibition, both mKATP channel openers differentially attenuated mitochondrial respiration. Neither mKATP channel antagonist abolished these effects. When ATP synthase was inhibited by oligomycin to decrease [ATP], both mKATP channel openers accelerated respiration for both substrate groups. This was abolished by mKATP channel blockade. Thus, under energetically more physiological conditions, the main effect of mKATP channel openers on mitochondrial respiration is differential inhibition independent of mKATP channel opening. In contrast, under energetically less physiological conditions, mKATP channel opening can be evidenced by accelerated respiration and blockade by antagonists. Therefore, the effects of mKATP channel openers on mitochondrial function likely depend on the experimental conditions and the cell\u27s underlying energetic state

Tyrosine Nitration of Voltage-dependent Anion Channels in Cardiac Ischemia-reperfusion: Reduction by Peroxynitrite Scavenging

Excess superoxide (O2−) and nitric oxide (NO) forms peroxynitrite (ONOO−) during cardiac ischemia reperfusion (IR) injury, which in turn induces protein tyrosine nitration (tyr-N). Mitochondria are both a source of and target for ONOO−. Our aim was to identify specific mitochondrial proteins that display enhanced tyr-N after cardiac IR injury, and to explore whether inhibiting O2−/ONOO− during IR decreases mitochondrial protein tyr-N and consequently improves cardiac function. We show here that IR increased tyr-N of 35 and 15 kDa mitochondrial proteins using Western blot analysis with 3-nitrotyrosine antibody. Immunoprecipitation (IP) followed by LC–MS/MS identified 13 protein candidates for tyr-N. IP and Western blot identified and confirmed that the 35 kDa tyr-N protein is the voltage-dependent anion channel (VDAC). Tyr-N of native cardiac VDAC with IR was verified on recombinant (r) VDAC with exogenous ONOO−. We also found that ONOO− directly enhanced rVDAC channel activity, and rVDAC tyr-N induced by ONOO− formed oligomers. Resveratrol (RES), a scavenger of O2−/ONOO−, reduced the tyr-N levels of both native and recombinant VDAC, while L-NAME, which inhibits NO generation, only reduced tyr-N levels of native VDAC. O2− and ONOO− levels were reduced in perfused hearts during IR by RES and L-NAME and this was accompanied by improved cardiac function. These results identify tyr-N of VDAC and show that reducing ONOO− during cardiac IR injury can attenuate tyr-N of VDAC and improve cardiac function

Flow-Based Single Cell Deposition for High-Throughput Screening of Protein Libraries

The identification and engineering of proteins having refined or novel characteristics is an important area of research in many scientific fields. Protein modelling has enabled the rational design of unique proteins, but high-throughput screening of large libraries is still required to identify proteins with potentially valuable properties. Here we report on the development and evaluation of a novel fluorescent activated cell sorting based screening platform. Single bacterial cells, expressing a protein library to be screened, are electronically sorted and deposited onto plates containing solid nutrient growth media in a dense matrix format of between 44 and 195 colonies/cm2. We show that this matrix format is readily applicable to machine interrogation (<30 seconds per plate) and subsequent bioinformatic analysis (~60 seconds per plate) thus enabling the high-throughput screening of the protein library. We evaluate this platform and show that bacteria containing a bioluminescent protein can be spectrally analysed using an optical imager, and a rare clone (0.5% population) can successfully be identified, picked and further characterised. To further enhance this screening platform, we have developed a prototype electronic sort stream multiplexer, that when integrated into a commercial flow cytometric sorter, increases the rate of colony deposition by 89.2% to 24 colonies per second. We believe that the screening platform described here is potentially the foundation of a new generation of high-throughput screening technologies for proteins

Prospects for precision measurements of atomic helium using direct frequency comb spectroscopy

We analyze several possibilities for precisely measuring electronic transitions in atomic helium by the direct use of phase-stabilized femtosecond frequency combs. Because the comb is self-calibrating and can be shifted into the ultraviolet spectral region via harmonic generation, it offers the prospect of greatly improved accuracy for UV and far-UV transitions. To take advantage of this accuracy an ultracold helium sample is needed. For measurements of the triplet spectrum a magneto-optical trap (MOT) can be used to cool and trap metastable 2^3S state atoms. We analyze schemes for measuring the two-photon $2^3S \to 4^3S$ interval, and for resonant two-photon excitation to high Rydberg states, $2^3S \to 3^3P \to n^3S,D$. We also analyze experiments on the singlet-state spectrum. To accomplish this we propose schemes for producing and trapping ultracold helium in the 1^1S or 2^1S state via intercombination transitions. A particularly intriguing scenario is the possibility of measuring the $1^1S \to 2^1S$ transition with extremely high accuracy by use of two-photon excitation in a magic wavelength trap that operates identically for both states. We predict a ``triple magic wavelength'' at 412 nm that could facilitate numerous experiments on trapped helium atoms, because here the polarizabilities of the 1^1S, 2^1S and 2^3S states are all similar, small, and positive.Comment: Shortened slightly and reformatted for Eur. Phys. J.

A Modified Approach to Single-Spin Detection Using Magnetic Resonance Force Microscopy

The magnetic moment of a single spin interacting with a cantilever in magnetic resonance force microscopy (MRFM) experiences quantum jumps in orientation rather than smooth oscillations. These jumps cannot be detected by a conventional MRFM based on observation of driven resonant oscillations of a cantilever. In this paper, we propose a method which will allow detection of the magnetic signal from a single spin using a modification of a conventional MRFM. We estimate the opportunity to detect the magnetic signal from a single proton.Comment: 4 pages LaTex, 4 figures in GIF forma

Velocity-selective direct frequency-comb spectroscopy of atomic vapors

We present an experimental and theoretical investigation of two-photon direct frequency-comb spectroscopy performed through velocity-selective excitation. In particular, we explore the effect of repetition rate on the $\textrm{5S}_{1/2}\rightarrow \textrm{5D}_{3/2, 5/2}$ two-photon transitions excited in a rubidium atomic vapor cell. The transitions occur via step-wise excitation through the $\textrm{5P}_{1/2, 3/2}$ states by use of the direct output of an optical frequency comb. Experiments were performed with two different frequency combs, one with a repetition rate of $\approx 925$ MHz and one with a repetition rate of $\approx 250$ MHz. The experimental spectra are compared to each other and to a theoretical model.Comment: 10 pages, 7 figure

Hydrogen Motion in Magnesium Hydride by NMR

In coarse-grained MgH2, the diffusive motion of hydrogen remains too slow (<10^5 hops s^−1) to narrow the H NMR line up to 400 °C. Slow-motion dipolar relaxation time T1D measurements reveal the motion, with hopping rate ωH from 0.1 to 430 s^−1 over the range of 260 to 400 °C, the first direct measurement of H hopping in MgH2. The ωH data are described by an activation energy of 1.72 eV (166 kJ/mol) and attempt frequency of 2.5 × 10^15 s^−1. In ball-milled MgH2 with 0.5 mol % added Nb2O5 catalyst, line-narrowing is evident already at 50 °C. The line shape shows distinct broad and narrow components corresponding to immobile and mobile H, respectively. The fraction of mobile H grows continuously with temperature, reaching ∼30% at 400 °C. This demonstrates that this material’s superior reaction kinetics are due to an increased rate of H motion, in addition to the shorter diffusion paths from ball-milling. In ball-milled MgH2 without additives, the line-narrowed component is weaker and is due, at least in part, to trapped H2 gas. The spin−lattice relaxation rates T1^−1 of all materials are compared, with ball-milling markedly increasing T1^−1. The weak temperature dependence of T1^−1 suggests a mechanism with paramagnetic relaxation centers arising from the mechanical milling

Coherence as ultrashort pulse train generator

Intense, well-controlled regular light pulse trains start to play a crucial role in many fields of physics. We theoretically demonstrate a very simple and robust technique for generating such periodic ultrashort pulses from a continuous probe wave which propagates in a dispersive thermal gas media

Temperature-Programmed Desorption: Principles, Instrument Design, and Demonstration With NaAlH4

This article is a brief introduction to temperature-programmed desorption (TPD), an analytical technique devised to analyze, in this case, materials for their potential as hydrogen storage materials. The principles and requirements of TPD are explained and the different components of a generic TPD apparatus are described. The construction of a modified TPD instrument from commercially available components is reported together with the control and acquisition technique used to create a TPD spectrum. The chemical and instrumental parameters to be considered in a typical TPD experiment and the analytical utility of the technique are demonstrated by the dehydrogenation of titanium-doped NaAlH{sub 4} by means of thermally programmed desorption