Modification of chirped laser pulses via delayed rotational nonlinearity.

To interpret single-shot measurements of rotational revival patterns in molecular gases excited by an ultrashort laser pulse, an analytical description of the probe pulse modulation by the impulsively excited medium is developed. A femtosecond pump laser pulse prepares a rotational wavepacket in a gas-phase sample, and the resulting periodic revivals are mapped into the frequency domain by using a substantially chirped continuum probe pulse. Since the standard approximate descriptions of probe pulse propagation are inapplicable (such as the slowly varying envelope approximation and the slowly evolving wave approximation), we propose an approach capable of incorporating both the substantial chirp of the pulse and the temporal dispersion of the medium response. Theory is presented for the case where the frequency change of the probe during the probe pulse duration is comparable with the carrier frequency. Analytical expressions are obtained for the probe signal modulation over the pump-probe interaction region and for the resulting heterodyned transient birefringence spectra. The approach is illustrated using the case of nitrogen gas.

Control of spectral interference patterns in broad Rabi sidebands toward quasi-comb structures.

The pattern of spectral interference fringes in broad dynamic Rabi sidebands allows for a considerable degree of control by shaping the picosecond driving pulse. We demonstrate experimental evidence of such control and report an analytic and numerical investigation of possibilities to control the fringe pattern to produce a comb-like optical structure. The temporal phase and amplitude shaping of a picosecond driving pulse influence the spectrum envelope, fringe contrast, and fringe spacing variation in the sideband spectra. The sideband spectrum envelope depends on the sharpness of the driving pulse, that is, on the rate at which the temporal distance between the leading and trailing edges grows away from the pulse maximum. Increasing this parameter reduces the variation of the envelope amplitude across the sideband. The fringe contrast, defined by the maximum-to-minimum difference, depends strongly on the asymmetry of the driving pulse. The imbalance between the leading and trailing edges leads to a decrease of the contrast. The variation of interpeak distance within a sideband was controlled using the temporal shape of the driving pulse. In the particular case of a blue-shifted sideband emitted by excited oxygen atoms driven by a picosecond pulse of 800 nm carrier wavelength and ∼5×10¹° W cm⁻² intensity, a Gaussian pulse shape results in an interpeak distance increasing almost five times over the interval from 1.60 to 1.66 eV, whereas a super-Gaussian shape leads to almost equidistant fringes producing a comb-like spectrum.

Postionization medium evolution in a laser filament: A uniquely nonplasma response.

Theoretical consideration of the optical response of nascent free electrons in the process of laser filamentation reveals that the initial microscopically inhomogeneous charge distribution causes a transient electromagnetic response of the medium that differs drastically from that of a homogeneous plasma with the same degree of ionization. An analytical model, describing the forced oscillations of virtually isolated and expanding electron clouds, predicts considerable enhancement of these oscillations caused by transient resonance with the laser field. The transient resonance processes should play a role in the currently accepted picture of filament formation dynamics.

Ionization-grating-induced nonlinear phase accumulation in spectrally resolved transient birefringence measurements at 400 nm.

We report experimental confirmation of the ionization-grating-induced transient birefringence predicted by Wahlstrand and Milchberg [Opt. Lett. 36, 3822 (2011)] and discuss its impact on the higher-order Kerr effect interpretation by Loriot et al. of pump-probe transient birefringence measurements made at 800 nm [Opt. Express 17, 13429 (2009)]. Measurement of the transient birefringence in air at 400 nm shows a negative contribution to the index of refraction at zero delay for frequencies within the pump bandwidth, the degenerate case, and no negative contribution for frequencies exceeding the pump bandwidth, the nondegenerate case. Our findings suggest that a reevaluation of the higher-order Kerr effect hypothesis of Loriot et al. is necessary.

Spatial-spectral distribution of Rabi radiation generated in plasma.

We report observation and control of the spatial-spectral distributions of coherent, dynamic Rabi sideband radiation. The Rabi sidebands result from the interaction of a shaped picosecond probe laser of intensity 10(10) W cm(-2), with neutral excited atomic oxygen generated in a laser-induced microplasma. The spatial-spectral distribution is measured and compared for picosecond laser pulses having either an asymmetric temporal shape or a Gaussian temporal shape. The resulting spatial-spectral distributions are in quantitative agreement with theoretical predictions that account for the radial intensity distribution of the picosecond probe pulse.

Self-shortening dynamics measured along a femtosecond laser filament in air.

The filamentation-induced temporal shortening of a 40 femtosecond pulse propagating in air is traced using impulsive vibrational Raman scattering and measurement of the power spectrum as a function of position along the propagation axis. The N2, O2, and H2O vibrational Raman responses reveal self-shortening of pulse features to 14 fs during the first filamentation cycle and to at least 9 fs in the second cycle. Spectral measurements further demonstrate that the coherent bandwidth generated in the region from 470 to 330 nm during the self-shortening process forms the ∼9 fs pulse.

Origin of the spectral coherence in dynamically broadened Rabi sidebands.

We demonstrate the coherent nature of dynamic Rabi-shifted sidebands arising when a picosecond probe laser interacts with a weakly ionized laser-generated microplasma. The coherence is manifested as spectral fringes observed in the sideband spectra. A model is presented that quantitatively predicts both the spectral phase and the spectral interference measured in the sidebands.

Rovibrational wave-packet dispersion during femtosecond laser filamentation in air.

An impulsive, femtosecond filament-based Raman technique producing high quality Raman spectra over a broad spectral range (1554.7-4155 cm(-1)) is presented. The temperature of gas phase molecules can be measured by temporally resolving the dispersion of impulsively excited vibrational wave packets. Application to laser-induced filamentation in air reveals that the initial rovibrational temperature is 300 K for both N2 and O2. The temperature-dependent wave-packet dynamics are interpreted using an analytic anharmonic oscillator model. The wave packets reveal a 1/e dispersion time of 3.9 ps for N2 and 2.8 ps for O2. Pulse self-compression of temporal features to 8 fs within the filament is directly measured by impulsive vibrational excitation of H2.

Impact-ionization cooling in laser-induced plasma filaments.

The ionization rates and subsequent electron dynamics for laser-induced plasma channels are measured for the noble gas series He, Ne, Ar, Kr, and Xe at 1.0 atm. The cw fluorescence emission increases superlinearly in the series from He to Xe in agreement with Ammosov-Delone-Krainov tunnel ionization calculations. The electron temperature after laser-induced plasma formation, measured by four-wave mixing, evolves from >20 eV to <1 eV kinetic energies with time constants ranging from 1 ns for He to 100 ps for Xe in agreement with an impact-ionization cooling model.

Elucidating the spectral and temporal contributions from the resonant and nonresonant response to femtosecond coherent anti-Stokes Raman scattering.

A theoretical expression is developed for femtosecond coherent anti-Stokes Raman scattering (CARS) to quantitatively account for the vibrational line shape in the presence of nonresonant signal. The contributions of the resonant and nonresonant components are extracted from the emitted signal line shape as a function of Stokes wavelength and as a function of the temporal overlap of the two pump pulses (for spectrally resolved femtosecond CARS). The theory is compared to the measured spectra of the oxygen vibrational transition DeltaG(01)=1556.4 cm(-1) for temporal detunings of 0 and 700 fs.

Phase matching in femtosecond BOXCARS.

We analytically describe the effect of phase-matching conditions on femtosecond BOXCARS line shape and demonstrate quantitative agreement with experimental spectra for the oxygen vibrational transition, DG01=1556.4 cm(-1).

Heterochromatic distribution of HeT-A- and TART-like sequences in several Drosophila species.

Drosophila melanogaster telomeres contain arrays of two non-LTR retrotransposons called HeT-A and TART. Previous studies have shown that HeT-A- and TART-like sequences are also located at non-telomeric sites in the Y chromosome heterochromatin. By in situ hybridization experiments, we mapped TART sequences in the h16 region of the long arm close to the centromere of the Y chromosome of D. melanogaster. HeT-A sequences were localized in two different regions on the Y chromosome, one very close to the centromere in the short arm (h18-h19) and the other in the long arm (h13-h14). To assess a possible heterochromatic location of TART and HeT-A elements in other Drosophila species, we performed in situ hybridization experiments, using both TART and HeT-A probes, on mitotic and polytene chromosomes of D. simulans, D. sechellia, D. mauritiana, D. yakuba and D. teissieri. We found that TART and HeT-A probes hybridize at specific heterochromatic regions of the Y chromosome in all Drosophila species that we analyzed.

Cells of human aminopeptidase N (CD13) transgenic mice are infected by human coronavirus-229E in vitro, but not in vivo.

Aminopeptidase N, or CD13, is a receptor for serologically related coronaviruses of humans, pigs, and cats. A mouse line transgenic for the receptor of human coronavirus-229E (HCoV-229E) was created using human APN (hAPN) cDNA driven by a hAPN promoter. hAPN-transgenic mice expressed hAPN mRNA in the kidney, small intestine, liver, and lung. hAPN protein was specifically expressed on epithelial cells of the proximal convoluted renal tubules, bronchi, alveolar sacs, and intestinal villi. The hAPN expression pattern within transgenic mouse tissues matched that of mouse APN and was similar in mice heterozygous or homozygous for the transgene. Primary embryonic cells and bone marrow dendritic cells derived from hAPN-transgenic mice also expressed hAPN protein. Although hAPN-transgenic mice were resistant to HCoV-229E in vivo, primary embryonic cells and bone marrow dendritic cells were infected in vitro. hAPN-transgenic mice are valuable as a source of primary mouse cells expressing hAPN. This hAPN-transgenic line will also be used for crossbreeding experiments with other knockout, immune deficient, or transgenic mice to identify factors, in addition to hAPN, that are required for HCoV-229E infection.

Fabrication of patch pipets.

Patch clamping refers to a wide range of electrophysiological measurements, all of which have in common the use of patch pipets and the formation of gigaohm seals. The purpose of this unit is to describe the fabrication of patch pipets. The aspects of the pipet geometry that are important to different applications and the different procedures that have been found to most reliably and simply achieve these results are described. Parameters for glass selection are detailed in the beginning of the unit. Pulling patch and whole-cell pipets, elastomer coating, fire polishing, pipet filling, and pipet testing in an experimental setup are highlighted. Additional support protocols describe alternative ways to optimize pipet geometry and cleaning the glass before pulling. Considerations for noise and dynamic performance are emphasized as these two requirements for single-channel and whole-cell current measurements dictate how the pipets must be fabricated.

Selective bond dissociation and rearrangement with optimally tailored, strong-field laser pulses.

We used strong-field laser pulses that were tailored with closed-loop optimal control to govern specified chemical dissociation and reactivity channels in a series of organic molecules. Selective cleavage and rearrangement of chemical bonds having dissociation energies up to approximately 100 kilocalories per mole (about 4 electron volts) are reported for polyatomic molecules, including (CH3)2CO (acetone), CH3COCF3 (trifluoroacetone), and C6H5COCH3 (acetophenone). Control over the formation of CH(3)CO from (CH3)2CO, CF3 (or CH3) from CH3COCF3, and C6H5CH3 (toluene) from C6H5COCH3 was observed with high selectivity. Strong-field control appears to have generic applicability for manipulating molecular reactivity because the tailored intense laser fields (about 10(13) watts per square centimeter) can dynamically Stark shift many excited states into resonance, and consequently, the method is not confined by resonant spectral restrictions found in the perturbative (weak-field) regime.

Infectious RNA transcripts from full-length dengue virus type 2 cDNA clones made in yeast.

The dengue virus type 2 genomic RNA was amplified by reverse transcription-PCR and cloned as four cDNA fragments. We could not assemble these four fragments into full-length cDNA in Escherichia coli. The full-length dengue virus cDNA was constructed by homologous recombination in yeast, either as part of a yeast artificial chromosome or in a yeast-E. coli shuttle vector. Full-length cDNA clones were propagated once in E. coli to prepare useful quantities of DNA. In vitro transcription of these clones produced full-length RNA transcripts. Introduction of these transcripts into LLC-MK2 cells produced typical dengue infection, as judged by cytopathic effects and indirect immunofluorescence. Infectivity was sensitive to RNase digestion and was dependent on the presence of cap analog in the transcription reaction mixture. Virus in the medium was passaged on C6-36 cells to produce stocks, and these stocks had titers and plaque morphologies similar to those of the parental dengue virus type 2. Intracellular dengue virus RNA from cells infected with transcript-derived virus contained an introduced BstEII site, proving that infectivity was derived from RNA transcripts and not from contamination with parental dengue virus. Transcript-derived virus was comparable to dengue virus type 2 for growth and protein expression in tissue culture cells. Sequence analysis of the dengue virus cDNA in one full-length clone revealed only one unexpected silent mutation. By using yeast technology, it will be easy to introduce specific mutations into the dengue virus cDNA, allowing analysis of the virus phenotype in cells transfected with mutant transcripts.

Feline aminopeptidase N serves as a receptor for feline, canine, porcine, and human coronaviruses in serogroup I.

Two members of coronavirus serogroup I, human respiratory coronavirus HCV-229E and porcine transmissible gastroenteritis virus (TGEV), use aminopeptidase N (APN) as their cellular receptors. These viruses show marked species specificity in receptor utilization, as HCV-229E can utilize human but not porcine APN, while TGEV can utilize porcine but not human APN. To determine whether feline APN could serve as a receptor for two feline coronaviruses in serogroup I, feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FeCV), we cloned the cDNA encoding feline APN (fAPN) by PCR from cDNA isolated from a feline cell line and stably expressed it in FIPV- and FeCV-resistant mouse and hamster cells. The predicted amino acid sequence of fAPN shows 78 and 77% identity with human and porcine APN, respectively. When inoculated with either of two biologically different strains of FIPV or with FeCV, fAPN-transfected mouse and hamster cells became infected and viral antigens developed in the cytoplasm. Infectious FIPV was released from hamster cells stably transfected with fAPN. The fAPN-transfected mouse and hamster cells were challenged with other coronaviruses in serogroup I including canine coronavirus, porcine coronavirus TGEV, and human coronavirus HCV-229E. In addition to serving as a receptor for the feline coronaviruses, fAPN also served as a functional receptor for each of these serogroup I coronaviruses as shown by development of viral antigens in the cytoplasm of infected mouse or hamster cells stably transfected with fAPN. In contrast, fAPN did not serve as a functional receptor for mouse hepatitis virus (MHV-A59), which is in serogroup II and utilizes mouse biliary glycoprotein receptors unrelated to APN. Thus, fAPN serves as a receptor for a much broader range of group I coronaviruses than human and porcine APNs. The human, porcine, and canine coronaviruses in serogroup I that are able to use fAPN as a receptor have previously been shown to infect cats without causing disease. Therefore, host factors in addition to receptor specificity apparently affect the virulence and transmissibility of nonfeline serogroup I coronaviruses in the cat.

Multiple receptor-dependent steps determine the species specificity of HCV-229E infection.

Human coronavirus (HCV)-229E causes disease only in humans and grows in human cells and in cells of other species that express recombinant human aminopeptidase N (hAPN), the receptor for HCV-229E. We compared the species specificity of HCV-229E infection with the species specificity of virus binding using immunofluorescence, assay of virus yields, fluorescence activated cell sorting and a monoclonal antibody directed against hAPN that blocks infection. We found that HCV-229E binds to intestinal brush border membranes (BBM) and to membranes of cell lines from cats, dogs, pigs, and humans, however the virus only infects two of these species. HCV-229E will not bind to BBM or to membranes from cell lines derived from hamster or mice. Animal coronaviruses related to HCV-229E, including FIPV, CCV, and TGEV bind to cell membranes from cats, dogs, cows, pigs and humans (but not mice), while each virus infects cells from only a subset of these species. Infectious genomic HCV-229E RNA, can infect cells of all of these species. These data suggest that the species-specificity of infection for this serogroup of coronaviruses is determined at the levels of virus binding and penetration. Since binding of viral spike glycoprotein to cellular receptors is not the only limiting factor, we suggest that one or more steps associated with virus penetration may determine the species specificity of infection with the HCV-229E serogroup of coronaviruses.