APPLICATION OF LASER-INDUCED BREAKDOWN CAVITATION BUBBLES FOR CELL LYSIS IN VITRO
Keywords:Cavitation bubble, Laser-induced breakdown, Cell lysis, Cell viability
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 highspeed
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.
laser used as an adjunct irrigant on clinical isolate of Enterococcus
Faecalis biofilm in vitro. Int J App Pharm 2018;9:103-6.
2. Iselinni C, Meidyawati R, Djauharie N. Effects of a 980- nm diode
laser´s activation of 2.5% NaOCl and 2% chlorhexidine antifungal
irrigation solutions on Candida albicans biofilms. Int J App Pharm
3. Hellman AN, Rau KR, Yoon HH, Venugopalan V. Biophysical response
to pulsed laser microbeam-induced cell lysis and molecular delivery. J
4. Rau KR, Guerra A. Investigation of laser-induced cell lysis using rime
resolved imaging. Appl Phy Lett 2004;84:2940-2.
5. Rau KR, Quinto-Su PA, Hellman AN, Venugopalan V. Pulsed laser
microbeam-induced cell lysis: Time-resolved imaging and analysis of
hydrodynamic effects. Biophys J 2006;91:317-29.
Fig. 4: Impact of the bubble on the cell layer. Comparison of
bubble size and wound (cell lysis region) induced by its impact.
Scale bars: white = 3 mm, gray = 1 mm
6. Brown RB, Audet J. Current techniques for single-cell lysis. J R Soc
Interface 2008;5 Suppl 2:S131-8.
7. Brennen CE. Cavitation and Bubble Dynamics. New York: Cambridge
University Press; 2015. p. 59-88.
8. Brujan EA, Nahen K, Schmidt P, Vogel A. Dynamics of laser-induced
cavitation bubbles near elastic boundary. J Fluid Mech 2001;433:251-
9. Jasikova D, Muller M, Kotek M, Kopecky V. The synchronized
force impact measurement and visualization of single cavitation
bubblegenerated with LIB. Int J Mech 2015;9:76-82.
10. Islam MS. A review on macroscale and microscale cell lysis methods.
11. Kennedy PK, Hammer DX, Rockwell BA. Laser-induced breakdown in
aqueous media. Prog Quantum Electron 1997;21:155-248.
12. Vogel A, Noack J, Nahen K, Theisen D, Busch S, Parlitz U. Energy
balance of optical breakdown in water at nanosecond to femtosecond
time scales. Appl Phys B 1999;68:271-80.
13. Brujan EA, Nahen K, Schmidt P, Vogel A. Dynamics of laser-induced
cavitation bubbles near an elastic boundary. J Fluid Mech 2001;133:251-81.
14. Compton JL, Hellman AN, Venugopalan V. Hydrodynamic determinants
of cell necrosis and molecular delivery produced by pulsed laser
microbeam irradiation of adherent cells. Biophys J 2013;105:2221-31.