For example, there is evidence that endocytosis plays a role in the uptake of agents during sonoporation. In addition to physical disruption of the cell membrane, cavitation has been shown to provoke other biological effects. The stresses on the cell membrane due to cavitation can result in physical damage to the cell membrane allowing for the passive diffusion of agents across. At higher acoustic pressures, inertial cavitation can occur, resulting in the formation of shock waves as well as вЂњmicrojetsвЂќ in the presence of a relatively rigid cell membrane. In, the authors captured high-speed images of oscillating microbubbles pushing and pulling on the cell membrane when in close proximity, an interaction which correlated with increased uptake of propidium iodide. At low acoustic intensities, stable cavitation has been shown to induce shear stress on the cell membrane by the generation of rapid flow in the surrounding medium, referred to as вЂњmicrostreamingвЂќ. Microbubble cavitation is known to induce a number of biophysical effects. In practice, clinically approved ultrasound contrast agents, comprised of microscale inert gas bubbles (вЂњmicrobubblesвЂќ) often having lipid, polymer, or protein shells, are added to cell media to serve as cavitation nuclei during ultrasound exposure. This process has been shown to be highly dependent on the presence of cavitation nuclei. Sonoporation is a process in which ultrasound exposure can induce a transient increase in the permeability of cell membranes. The prototype device was found to be suitable for performing sonoporation on adherent cell cultures and will reduce the time and expertise required for conducting sonoporation experiments on adherent cell cultures in the future. ConclusionsД«ath temperature did not impact the efficacy of sonoporation treatment on SiHa cells in vitro. Neither GFP expression, PARP expression, or detachment differed significantly between bath temperatures. GFP expression and cell detachment were both significantly affected by the presence of ultrasound contrast agent ( p < 0.001, p < 0.001).

Cell detachment was measured using phase contrast microscopy before and after treatment. GFP expression and PARP expression following treatment were measured using fluorescent microscopy and digital image processing. Each treatment was performed in biological triplicate. Ultrasound treatment with Definity ultrasound contrast agent (US+UCA) present, ultrasound treatment without contrast agent present (US), and a sham ultrasound treatment in the presence of ultrasound contrast agent (CA) were each performed at bath temperatures of 37, 39.5, and 42 вЊ. A near field treatment plane was chosen for each transducer to minimize field heterogeneity in the near field.Д¬ervical cancer-derived SiHa cells were exposed to nine different treatments in the presence of plasmid DNA-expressing green fluorescent protein (GFP). Six transducers with nominal diameters of 20 mm were constructed and the acoustic field of each was characterized using hydrophone scanning. The prototype device consisted of six ultrasound transducers affixed beneath an Opticell stage. This prototype device was used to examine the superficial effect of bath temperature on sonoporation efficacy. A device was devised which aimed to reduce the time and expertise required to perform sonoporation on adherent cell cultures.

Cellprofiler minimizing contrast