Eclipse 80i upright microscope

The acoustic cell setup consisted of a piezoelectric transducer
(Ferroperm PZ26 disk 10 mm diameter, 0.5 mm thickness) responsible for the
generation of ultrasound waves, attached by conductive epoxy glue to the bottom
center of a steel plate (50 mm × 50 mm × 0.94 mm³);
then the steel plate was covered with a 240 µm kapton tape protective
layer and a sample reservoir at the center (5 mm). A glass slide was used to
cover the reservoir for ultrasound reflection and to protect the sample. The
continuous ultrasound sine wave was applied via a piezoelectric
transducer, through an Agilent 15 MHz arbitrary waveform generator, in
connection to a home-made power amplifier. The applied continuous sine wave form
had a frequency of 2.56 MHz and 6 V voltage amplitude.
Videos were captured using Cool SNAP HQ² camera, 20× and 40× objectives
(unless mentioned otherwise) and acquired at the frame rate of 10 using the
Metamorph 7.1 software (Molecular Devices, Sunnyvale, CA). A Nikon Eclipse 80i
upright microscope was also used to capture time course images of the
morphological changes of AGS cells treated with US-propelled pH-responsive
polymer-CASP-3@nanomotors and other conditions.

Dissected mouse tissues were embedded in OCT media, snap frozen, and stored at −80°C. Sections (10 μm) were cut using a Micron HM505 N cryostat, placed onto glass slides, air dried for 10 min, and then fixed in 4% paraformaldehyde for 10 min. Slides were washed twice in cold 1× PBS buffer and then blocked with the appropriate serum-blocking buffer at room temperature for 1 h. Slides were incubated overnight at 4°C with primary anti-Zhx2 antibody (1:250; #96083; Abcam), washed twice with 1× PBS, and incubated with TRITC-conjugated secondary antibody (1:200; #4010-13; Southern Biotech) for 1.5 h at room temperature. Slides were mounted (Dako mounting media; #2013-05), covered, and imaged with a Nikon Eclipse 80i upright microscope with NiS Elements software.