Abstract
Objective: Understanding the basic mechanism of the cavitation bubble action on living cells as a crucial step of development and application ofsophisticated methods based on controlled cavitation in cell behaviour manipulation. Optimisation of parameters in order to expand cell lysis regioncreated by a single bubble.Methods: The cavitation bubbles are generated by the laser-induced breakdown method. The impact of controlled cavitation bubble on thebiological system is synchronously monitored under a microscope and recorded. Visualization of the cavitation bubble course is monitored by a highspeedcamera. The impact of technology on the healthy confluent cell layer is verified. Evaluation of the cavitation bubbles´ effect on cells in real timeand by subsequent analysis of the cell lysis region and impact of the cavitation bubble on cell viability is carried out by optical visualization and life/dead fluorescence staining.Results: Cavitation bubble induced in distance of 1.5 mm from the cell surface overcomes properties of sessile bubble and enables to create cell lysisregion over 1000 μm in diameter due to transient shear stress produced by liquid displaced by the bubble expansion.Conclusion: Cell lysis region is strongly dependent on the spot laser energy (SLE) and the bubble induction distance from cells. This knowledge iscrucial for application in chemical free cell lysis in vitro, wound induction for experimental purposes and cell layers patterning in desired scale.
Highlights
The current trend in biotechnologies, including drug delivery, wound cleaning, and biofilm management is chemical-free and environmentally friendly processing based on cell lysis
The experiment was designed for synchronous recording of the evolution and collapse of the cavitation bubble generated by the laser-induced breakdown (LIB) device, while at the same time the impact zone in the cell layer was monitored using an inverted microscope
In contrary to the results of Hellman et al [3], who during their experiment produced targeted single cells and cell lysis regions of up to 300 μm surrounded by viable cells, we found that the distance modification combined with optimal spot laser energy (SLE) leads to the formation of a cavitation bubble with a much wider and more complex impact zone
Summary
The current trend in biotechnologies, including drug delivery, wound cleaning, and biofilm management is chemical-free and environmentally friendly processing based on cell lysis. One of the methods used to generate the cavitation bubble is laser-induced breakdown (LIB) [7,8,9]. Among all of the cell lysis methods, laser-induced cavitation possess several advantages, such as its independence of cell type, its operation at lower temperatures, and its absence of chemical agents interacting problematically with some biochemical methods/compounds. The LIB generation mechanism through microscope lenses leads to the formation of cavitation bubbles of micrometer sizes. The advantage of these methods is the use of very low levels of energy (μJ) in femtosecond pulses to create a cavitation bubble in close proximity to the cell substrate, which is known as a sessile bubble. The defects induced by the cavitation microbubbles affect a very small area on the cell substrate, the necrotic area of the cells can reach a diameter of up to 150 μm [3]
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