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APPLICATION OF LASER-INDUCED BREAKDOWN CAVITATION BUBBLES FOR CELL LYSIS IN VITRO
摘要： Objective: Understanding the basic mechanism of the cavitation bubble action on living cells as a crucial step of development and application of sophisticated methods based on controlled cavitation in cell behaviour manipulation. Optimisation of parameters in order to expand cell lysis region created by a single bubble. Methods: The cavitation bubbles are generated by the laser-induced breakdown method. The impact of controlled cavitation bubble on the biological system is synchronously monitored under a microscope and recorded. Visualization of the cavitation bubble course is monitored by a high-speed camera. The impact of technology on the healthy confluent cell layer is verified. Evaluation of the cavitation bubbles′ effect on cells in real time and 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 lysis region 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 is crucial for application in chemical free cell lysis in vitro, wound induction for experimental purposes and cell layers patterning in desired scale.
关键词： Laser-induced breakdown,Cavitation bubble,Cell viability,Cell lysis
更新于2025-09-12 10:27:22
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Micropatterning of Porous Silicon Bragg Reflectors With Poly(Ethylene Glycol) to Fabricate Cell Microarrays: Towards Single Cell Sensing
摘要： The work presented here describes the development of optical label-free biosensor based on PSi Bragg reflector to study heterogeneity in single cells. Photolithographic patterning of poly(ethylene glycol) hydrogel with photoinitiator was employed on RGD peptide-modified PSi to create micropatterns with cell adhesive and cell repellent areas and J774 macrophage cells were incubated to form cell microarrays and single cell arrays. Moreover, cells on the microarrays were lysed osmotically with Milli-Q? water and the infiltration of cell lysate into the porous matrix was monitored by measuring the red shift in the reflectivity. On average, the magnitude of red shift increased with the increase in the number of cells on the micropatterns. The red shift from the spots with single cells varied from spot to spot emphasizing the heterogeneous nature of the individual cells.
关键词： porous silicon (PSi),cell patterning,cell lysis,label-free biosensing,poly(ethylene glycol (PEG) hydrogel
更新于2025-09-04 15:30:14