Methods
Summary
A small hole is cut in the IV tubing. PDMS mixed in a 1:10 ratio is applied to the edges of this whole. 2.5 μm mylar film, which is acoustically transparent is applied to patch the hole. The tube will be placed in an oven for 24 hours at 60 C so the PDMS can cure. A high-pressure vacuum chamber at 300 kPa is used to degas water for 24 hours for the acrylic tank, and a water heater heats it to 37 C. A custom 3D-printed rig holds the IV tube right above the surface of the degassed water. The XYZ Motor System aligns the transducer with the Fiber Optic Hydrophone underneath the rig. The tube will be placed back on the rig and the IV syringe pump will be turned on. The generator will be controlled to generate LIPUS signals while the blood cancer cells flow through the tube. Cell viability is estimated using an apoptosis kit under a plate reader.
Cells are cultured in RPMI Media(10% FBS, 5% penicillin/streptomycin) and kept at 37C with 5% CO2.
See past Full Microplate Protocol
Challenges
Liquids may leak out of the IV tube at first. Sterile saline will be used first to test the structural integrity. If leaks are found, additional PDMS will be applied for future tubes
Pre Analysis Plan
An apoptosis kit will be used to estimate cell death using the ELISA method. Chemical interactions between the test-kit and intra-cellular markers will result in fluorescence. A microplate plate reader with a 450 nm lens will record the fluorescence for analysis in both treated/untreated and cancer/healthy cell groups.
Protocols
Browse the protocols that are part of the experimental methods.