Cytokine receptor-Eb1 interaction couples cell polarity and fate during asymmetric cell division.

Asymmetric stem cell division is a critical mechanism for balancing self-renewal and differentiation. Adult stem cells often orient their mitotic spindle to place one daughter inside the niche and the other outside of it to achieve asymmetric division. It remains unknown whether and how the niche may direct division orientation. Here we discover a novel and evolutionary conserved mechanism that couples cell polarity to cell fate. We show that the cytokine receptor homolog Dome, acting downstream of the niche-derived ligand Upd, directly binds to the microtubule-binding protein Eb1 to regulate spindle orientation in Drosophila male germline stem cells (GSCs). Dome's role in spindle orientation is entirely separable from its known function in self-renewal mediated by the JAK-STAT pathway. We propose that integration of two functions (cell polarity and fate) in a single receptor is a key mechanism to ensure an asymmetric outcome following cell division.

Noninvasive chorioretinal imaging in living rabbits using integrated photoacoustic microscopy and optical coherence tomography.

Most reported photoacoustic ocular imaging work to date uses small animals, such as mice and rats, the eyeball sizes of which are less than one-third of those of humans, posing challenges for clinical translation. Here we developed a novel integrated photoacoustic microscopy (PAM) and optical coherence tomography (OCT) system for dual-modality chorioretinal imaging of larger animals, such as rabbits. The system has quantified lateral resolutions of 4.1 µm (PAM) and 3.8 µm (OCT), and axial resolutions of 37.0 µm (PAM) and 4.0 µm (OCT) at the focal plane of the objective. Experimental results in living rabbits demonstrate that the PAM can noninvasively visualize individual depth-resolved retinal and choroidal vessels using a laser exposure dose of ~80 nJ below the American National Standards Institute (ANSI) safety limit 160 nJ at 570 nm; and the OCT can finely distinguish different retinal layers, the choroid, and the sclera. This reported work may be a major step forward in clinical translation of the technology.

High-precision, non-invasive anti-microvascular approach via concurrent ultrasound and laser irradiation.

Antivascular therapy represents a proven strategy to treat angiogenesis. By applying synchronized ultrasound bursts and nanosecond laser irradiation, we developed a novel, selective, non-invasive, localized antivascular method, termed photo-mediated ultrasound therapy (PUT). PUT takes advantage of the high native optical contrast among biological tissues and can treat microvessels without causing collateral damage to the surrounding tissue. In a chicken yolk sac membrane model, under the same ultrasound parameters (1 MHz at 0.45 MPa and 10 Hz with 10% duty cycle), PUT with 4 mJ/cm2 and 6 mJ/cm2 laser fluence induced 51% (p = 0.001) and 37% (p = 0.018) vessel diameter reductions respectively. With 8 mJ/cm2 laser fluence, PUT would yield vessel disruption (90%, p < 0.01). Selectivity of PUT was demonstrated by utilizing laser wavelengths at 578 nm or 650 nm, where PUT selectively shrank veins or occluded arteries. In a rabbit ear model, PUT induced a 68.5% reduction in blood perfusion after 7 days (p < 0.001) without damaging the surrounding cells. In vitro experiments in human blood suggested that cavitation may play a role in PUT. In conclusion, PUT holds significant promise as a novel non-invasive antivascular method with the capability to precisely target blood vessels.

Dual-pore glass chips for cell-attached single-channel recordings.

While high-throughput planar patch-clamp instruments are now established to perform whole-cell recordings for drug screening, the conventional micropipette-based approach remains the gold standard for performing cell-attached single-channel recordings. Generally, planar platforms are not well-suited for such studies due to excess noise resulting from low seal resistances and the use of substrates with poor dielectric properties. Since these platforms tend to use the same pore to position a cell by suction and establish a seal, biological debris from the cell suspension can contaminate the pore surface prior to seal formation, reducing the seal resistance. Here, femtosecond laser ablation was used to fabricate dual-pore glass chips optimized for use in cell-attached single-channel recordings that circumvent this problem by using different pores to position a cell and to establish a seal. This dual-pore design also permitted the use of a relatively small patch aperture (D ~ 150 to 300 nm) that is better-suited for establishing high-resistance seals than the micropores used typically in planar patch-clamp setups (D ~ 1 to 2 µm) without compromising the ability of the device to position a cell. Taking advantage of the high seal resistances and low capacitive and dielectric noise realized using glass substrates, patch-clamp experiments with these dual-pore chips consistently achieved high seal resistances (rate of gigaseal formation = 61%, mean seal resistance = 53 GΩ), maintained gigaseals for prolonged durations (up to 6 hours), achieved RMS noise values as low as 0.46 pA at 5 kHz bandwidth, and enabled single-channel recordings in the cell-attached configuration that are comparable to those obtained by conventional patch-clamp.

Pulsetrain-burst mode, ultrafast-laser interactions with 3D viable cell cultures as a model for soft biological tissues.

A 3D living-cell culture in hydrogel has been developed as a standardized low-tensile-strength tissue proxy for study of ultrafast, pulsetrain-burst laser-tissue interactions. The hydrogel is permeable to fluorescent biomarkers and optically transparent, allowing viable and necrotic cells to be imaged in 3D by confocal microscopy. Good cell-viability allowed us to distinguish between typical cell mortality and delayed subcellular tissue damage (e.g., apoptosis and DNA repair complex formation), caused by laser irradiation. The range of necrosis depended on laser intensity, but not on pulsetrain-burst duration. DNA double-strand breaks were quantified, giving a preliminary upper limit for genetic damage following laser treatment.

High-order harmonic generation from solid targets with 2 mJ pulses.

Harmonics up to the 18th order are generated from solid targets by focusing 2 mJ, 50 fs pulses at 800 nm to a spot size of 1.7 µm (FWHM). To our knowledge, this is the first demonstration of high-harmonic generation with a very short focal length paraboloid (f/1.4) and kilohertz laser system. The harmonics have a low divergence (<4°) compared to the driving beam and conversion efficiencies (>10(-7) per harmonic) comparable to gas harmonics. No contrast enhancement techniques are employed, and the system is capable of operating at 500 Hz.

Quasimonoenergetic electron beams with relativistic energies and ultrashort duration from laser-solid interactions at 0.5 kHz.

We investigate the production of electron beams from the interaction of relativistically-intense laser pulses with a solid-density SiO(2) target in a regime where the laser pulse energy is approximately mJ and the repetition rate approximately kHz. The electron beam spatial distribution and spectrum were investigated as a function of the plasma scale length, which was varied by deliberately introducing a moderate-intensity prepulse. At the optimum scale length of lambda/2, the electrons are emitted in a collimated beam having a quasimonoenergetic distribution that peaked at approximately 0.8 MeV. A highly reproducible structure in the spatial distribution exhibits an evacuation of electrons along the laser specular direction and suggests that the electron beam duration is comparable to that of the laser pulse. Particle-in-cell simulations which are in good agreement with the experimental results offer insights on the acceleration mechanism by the laser field.

Vacuum-free x-ray source based on ultrashort laser irradiation of solids.

A vacuum-free ultrafast laser-based x-ray source is demonstrated. Hard x-rays up to 80KeV are generated from Cu, Mo, Ag, Sn, and Ge targets in a laminar helium flow surrounded by atmosphere using tightly focused 33fs, 3mJ laser pulses. X-ray spectra, conversion efficiencies, and source sizes are presented. Six-fold efficiency improvement is observed, over similar sources found in the literature [1]. Source sizes determined for Cu and Mo show distinct dependences on laser pulse energy. It is also shown that the Cu source size has no dependence on the presence of the spectral band around the 8KeV K-shell lines.

High power fiber laser driver for efficient EUV lithography source with tin-doped water droplet targets.

In this paper we report the development of nanosecond-pulsed fiber laser technology for the next generation EUV lithography sources. The demonstrated fiber laser system incorporates large core fibers and arbitrary optical waveform generation, which enables achieving optimum intensities and other critical beam characteristics on a laser-plasma target. Experiment demonstrates efficient EUV generation with conversion efficiency of up to 2.07% for in-band 13.5-nm radiation using mass-limited Sn-doped droplet targets. This result opens a new technological path towards fiber laser based high power EUV sources for high-throughput lithography steppers.

Generation of hard X-rays using an ultrafast fiber laser system.

We report the first hard X-ray source driven by a femtosecond fiber laser. The high energy fiber CPA system incorporated a 65mum LMA fiber amplifying stage which provided 300-fs recompressed pulses and diffraction limited beam quality with M(2) < 1.07. A deformable mirror was used to optimize the wavefront and the spot size was focused down to 2.3 mum with an f/1.2 paraboloidal mirror. 50muJ was deposited on the nickel target with 2x10(15)-W/cm(2) focal intensity and a distinctive Ni K(alpha)-line (7.48 keV) emission was measured with 5x10(-8) energy conversion efficiency.

Demonstration of fiber-laser-produced plasma source and application to efficient extreme UV light generation.

Efficient generation of extreme UV (EUV) light at lambda = 13.5 nm from a bulk Sn target has been demonstrated by using a fiber laser. The conversion efficiency from the 1064 nm IR to the EUV was measured to be around 0.9% into 2pi steradians within a 2% bandwidth. To the best of our knowledge, this is the first time an all-fiber system was used to generate EUV or soft x rays.