Aroshan Jayasinghe

Apical oscillations in amnioserosa cells: basolateral coupling and mechanical autonomy.

Holographic laser microsurgery is used to isolate single amnioserosa cells in vivo during early dorsal closure. During this stage of Drosophila embryogenesis, amnioserosa cells undergo oscillations in apical surface area. The postisolation behavior of individual cells depends on their preisolation phase in these contraction/expansion cycles: cells that were contracting tend to collapse quickly after isolation; cells that were expanding do not immediately collapse, but instead pause or even continue to expand for ∼40 s. In either case, the postisolation apical collapse can be prevented by prior anesthetization of the embryos with CO2. These results suggest that although the amnioserosa is under tension, its cells are subjected to only small elastic strains. Furthermore, their postisolation apical collapse is not a passive elastic relaxation, and both the contraction and expansion phases of their oscillations are driven by intracellular forces. All of the above require significant changes to existing computational models.

Raman-shifted alexandrite laser for soft tissue ablation in the 6- to 7-µm wavelength range

Prior work with free-electron lasers (FELs) showed that wavelengths in the 6- to 7-µm range could ablate soft tissues efficiently with little collateral damage; however, FELs proved too costly and too complex for widespread surgical use. Several alternative 6- to 7-µm laser systems have demonstrated the ability to cut soft tissues cleanly, but at rates that were much too low for surgical applications. Here, we present initial results with a Raman-shifted, pulsed alexandrite laser that is tunable from 6 to 7 µm and cuts soft tissues cleanly—approximately 15 µm of thermal damage surrounding ablation craters in cornea—and does so with volumetric ablation rates of 2–5 × 10−3 mm3/s. These rates are comparable to those attained in prior successful surgical trials using the FEL for optic nerve sheath fenestration.

Holographic UV laser microsurgery.

Abstract We use a spatial light modulator (SLM) to diffract a single UV laser pulse to ablate multiple points on a Drosophila embryo. This system dynamically generates a phase hologram for ablating a user-defined pattern fast enough to be used with living, and thus moving, tissue. We demonstrate the ability of this single-pulse multi-point system to perform two experiments that are very difficult for conventional microsurgery—isolating single cells in vivo and measuring fast retractions from large incisions.

Interplay of wavelength, fluence and spot-size in free-electron laser ablation of cornea

Infrared free-electron lasers ablate tissue with high efficiency and low collateral damage when tuned to the 6-microm range. This wavelength-dependence has been hypothesized to arise from a multi-step process following differential absorption by tissue water and proteins. Here, we test this hypothesis at wavelengths for which cornea has matching overall absorption, but drastically different differential absorption. We measure etch depth, collateral damage and plume images and find that the hypothesis is not confirmed. We do find larger etch depths for larger spot sizes--an effect that can lead to an apparent wavelength dependence. Plume imaging at several wavelengths and spot sizes suggests that this effect is due to increased post-pulse ablation at larger spots.