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.

We developed a novel localized antivascular method, namely photo-mediated ultrasound therapy (PUT), by applying synchronized laser and ultrasound pulses. PUT relies on high optical contrast among biological tissues. Taking advantage of the high optical absorption of hemoglobin, PUT can selectively target microvessels without causing unwanted damages to the surrounding tissue. Moreover, PUT working at different optical wavelengths can selectively treat veins or arteries by utilizing the contrast in the optical spectra between deoxy- and oxy-hemoglobin. Through our experiments, we demonstrated that cavitation might have played a key role in PUT. The addition of a laser pulse to an existing ultrasound field can significantly improve the likelihood of inertial cavitation, which can induce microvessel damage through its mechanical effect. In comparison with conventional laser therapies, such as photothermolysis and photocoagulation, the laser energy level needed in PUT is significantly lower. When a nanosecond laser was used, our in vivo experiments showed that the needed laser fluence was in the range of 4 to 40 mJ/cm2.

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.

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.

Retinal and choroidal neovascularization play a pivotal role in the leading causes of blindness including macular degeneration and diabetic retinopathy (DR). Current therapy by focal laser photocoagulation can damage the normal surrounding cells, such as the photoreceptor inner and outer segments which are adjacent to the retinal pigment epithelium (RPE), due to the use of high laser energy and millisecond pulse duration. Therapies with pharmaceutical agents involve systemic administration of drugs, which can cause adverse effects and patients may become drug-resistant. We have developed a noninvasive photo-mediated ultrasound therapy (PUT) technique as a localized antivascular method, and applied it to remove micro blood vessels in the retina. PUT takes advantage of the high native optical contrast among biological tissues, and has the unique capability to self-target microvessels without causing unwanted damages to the surrounding tissues. This technique promotes cavitation activity in blood vessels by synergistically applying nanosecond laser pulses and ultrasound bursts. Through the interaction between cavitation and blood vessel wall, blood clots in microvessels and vasoconstriction can be induced. As a result, microvessels can be occluded. In comparison with other techniques that involves cavitation, both laser and ultrasound energy needed in PUT is significantly lower, and hence improves the safety in therapy.

In this work, a new therapeutic technique targeting blood vessels based on photo-mediated ultrasound therapy (PUT) was evaluated. By synergistically applying ultrasound waves on the target tissue during laser ablation, the cavitation effect can be greatly enhanced. This technique was applied to remove small blood vessels in a chicken embryo model and demonstrated promising outcomes. The treated blood vessels were either completely removed or significantly shrunk in size. In the control experiment where samples were treated with either ultrasound or laser alone, no changes in blood vessels were observed. With the advantages including high selectivity, high efficiency and label free, the developed PUT technique could be a new tool for treating a variety of diseases involving abnormal vasculature.

Second harmonic generation in scleral tissues is used to infer local tissue scattering and nonlinear susceptibility. Measurements from ex vivo porcine samples suggest that the femtosecond-laser threshold for subsurface photodisruption depends on both these properties.

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.

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.

Laser plasma interaction experiments have been performed to characterize high order harmonic emission up to the 18th order using high rep rate mJ level laser pulses at relativistic intensities. The experiments were compared to two- and three-dimensional particle-in-cell simulations. The harmonic divergence was found to be less than 4 degrees (full-width at half-maximum) at highest intensity and increased as the laser was defocused (i.e. as the intensity was reduced). The polarization dependence on the harmonic generation efficiency and divergence was also measured. Circular polarization was found to cause a deflection in the angle of emission of the harmonics-an effect which may be beneficial in the use of such harmonics for efficient isolated attosecond pulse production.

Laser plasma interaction reveals harmonic divergence <4 degrees and increasing with defocus. Circular polarization was found to cause a change in the angle of emission, which may be beneficial in efficient isolated attosecond pulse production.

We characterize the cellular impact of intense 1-ps laser irradiation at >100MHz repetition rates on viable cell cultures at several incident laser parameters (i.e., pulse intensity and burst duration).

AbstractUltrafast laser pulses (≤1 ps) are qualitatively different in the nature of their interaction with materials, including biotissues, as compared to nanosecond or longer pulses. This can confer pronounced advantages in outcomes for tissue therapy or laser surgery. At the same time, there are distinct limitations of their strong-field mode of interaction. As an alternative, it is shown here that ultrafast laser pulses delivered in a pulse-train burst mode of radiant exposure can access new degrees of control of the interaction process and of the heat left behind in tissues. Using a laser system that delivers 1 ps pulses in 20 μs pulse-train bursts at 133 MHz repetition rates, a range of heat and energy-transfer effects on hard and soft tissue have been studied. The ablation of tooth dentin and enamel under various conditions, to assess the ablation rate and characterize chemical changes that occur, are reported. This is compared to ablation in agar gels, useful live-cell-culture phantom of soft tissues, and presenting different mechanical strength. Study of aspects of the optical science of laser-tissue interaction promises to make qualitative improvements to medical treatments using lasers as cutting and ablative tools.

We've developed a 3-dimensional hydrogel cell culture to investigate the effects of ultrafast laser pulses on soft biological tissues. We characterize the physical and histological impact of intense laser irradiation at >;100MHz repetition rates.

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.

The energy spectrum of hot electrons emitted from the interaction of a relativistically intense laser with an Al plasma is measured at a repetition rate of 0.5 kHz by accumulating ∼103 highly reproducible laser shots. In the 1017–2×1018 W/cm2 range, the temperature of electrons escaping the plasma along the specular direction scales as (Iλ2)0.64±0.05 for p-polarized pulses incident at 45°. This scaling is in good agreement with three-dimensional particle-in-cell simulations and a simple model that estimates the hot-electron temperature by considering the balance between the deposited laser intensity and the energy carried away by those electrons.