Abstract
Laser-generated focused ultrasound (LGFU) is a unique modality that can produce single-pulsed cavitation and strong local disturbances on a tight focal spot (<100 μm). We utilize LGFU as a non-contact, non-thermal, high-precision tool to fractionate and cleave cell clusters cultured on glass substrates. Fractionation processes are investigated in detail, which confirms distinct cell behaviors in the focal center and the periphery of LGFU spot. For better understanding of local disturbances under LGFU, we use a high-speed laser-flash shadowgraphy technique and then fully visualize instantaneous microscopic processes from the ultrasound wave focusing to the micro-bubble collapse. Based on these visual evidences, we discuss possible mechanisms responsible for the focal and peripheral disruptions, such as a liquid jet-induced wall shear stress and shock emissions due to bubble collapse. The ultrasonic micro-fractionation is readily available for in vitro cell patterning and harvesting. Moreover, it is significant as a preliminary step towards high-precision surgery applications in future.
Highlights
Focused ultrasound with high intensity or high peak pressure can produce localized disruptions in terms of acoustic cavitation, streaming, and heat deposition [1,2]
We utilize Laser-generated focused ultrasound (LGFU) as a non-contact, non-thermal, high-precision tool to fractionate and cleave cell clusters cultured on glass substrates
Fractionation processes are investigated in detail, which confirms distinct cell behaviors in the focal center and the periphery of LGFU spot
Summary
Focused ultrasound with high intensity or high peak pressure can produce localized disruptions in terms of acoustic cavitation, streaming, and heat deposition [1,2] These effects have been broadly utilized for non-contact therapeutic applications such as shockwave lithotripsy [3,4], hyperthermia-based tumor treatment [5,6,7], and thrombolysis [8,9]. The cavitational impacts, together with shock-induced effects, have offered great potentials for in vitro cellular engineering in terms of selective cell detachment, patterning, and harvesting for cell-based assays and secondary analyses [13,14,15,16] Most of these ultrasonic disruptions were available over a bulky focal dimension (typically several mm) due to low operation frequencies (a few MHz) of existing high-pressure transducers [6]. Such dimension is unsuitable for performing micro-scale therapies and cellular engineering, and for exploring microscopic interaction mechanisms with cells in a new regime
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