How soundwaves trigger immune responses to cancer in mice umich frontiersin
). Briefly, histotripsy ablation was performed using 50 histotripsy pulses at 100 Hz pulse repetition frequency delivered at each location to generate an estimated −30 MPa peak negative pressure at the focus. Histotripsy ablation times ranged from 4-15 minutes. Tumor irradiation was performed by placing mice in radiation chambers with lead body shielding leaving only the tumor exposed to 15 Gy unfractionated radiation.
Samples were then washed and counterstained with Alexa 555-labeled goat anti-rabbit and Alexa 647 goat anti-mouse secondary antibodies at 37°C for 45 minutes. After washing, samples were incubated with mouse anti-HMGB1 antibody at 37°C for 1 hour. Samples were then washed and blocked in goat serum, incubated with Alexa 555-conjugated goat anti-rabbit secondary antibody for 45 minutes, and washed in PBS for 30 minutes.