JoVE Encyclopedia of Experiments
Cancer Research
0 views • 3:22 min • April 30th, 2023
- To begin, seed the human leukemia cells in a freshly prepared culture medium and incubate for the desired period. After incubation, stain a small amount of this cell suspension with trypan blue and assess cell viability using a TC20 cell counter slide. Dead cells are permeable and take up the negatively charged dye, while viable cells are impermeable.
Next, add methyl beta cyclodextrin to the culture flask to sufficiently deplete the cell's cholesterol, making them susceptible to ultrasound. Incubate the cells for the desired period. Fill the cell sonication system with deionized water and degas it for a few minutes to avoid vacuum bubbles. Transfer the desired amount of cells in the scintillation vial and attach the vial to the cell sonication system's holding device.
Place the holding device on top of the horn and set the desired elevation from the horn's top. Set required pulses for the desired duration to sonicate the cells. Add trypan blue to the cell suspension and place it in the TC20 cell counter to analyze cell viability and damage. Methyl beta cyclodextrin-treated leukemia cells, when exposed to low frequency ultrasound, show significant cell lysis.
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