Bright field inverted light microscope

The MDA-MB-453 and HCC-1937 cells were grown on coverslips (1 × 10⁵ cells/cover slip) and incubated with different concentrations (0.5, 1, or 1.5 mg/mL) of AME or with 0.1% DMSO for 48 h, and then fixed in a solution of ethanol: acetic acid (3:1, v/v). The cover slips were mounted on glass slides for morphometric analysis. Three monolayers per experimental group were photographed. Morphological changes in MDA-MB-453 and HCC-1937 cells were determined using a Nikon bright-field inverted light microscope (Tokyo, Japan) at 200× (for MDA-MB-453 cells) or 400× (for HCC-1937 cells) magnification.

Fat bodies were dissected in the precooled phosphate buffer saline, and the adherent fat bodies were carefully removed with forceps as thoroughly as possible under a stereomicroscope (SZM-7045, Nikon). The fat bodies from different time periods (every 7 days) were washed twice with 1× PBS buffer fixed with 4% paraformaldehyde for 30 min at room temperature. It was then washed twice with 1× PBS buffer. The fat bodies were then incubated in Nile Red solution (1 mg/ml Nile Red was diluted to 1 ug/mL by 0.1% Triton X-100) for 90 min at room temperature to stain the lipid droplets. Then it was washed twice with 1× PBS buffer. The cell nuclei were stained with 1 μg/ml DAPI (Sigma, D8417, United States) for an additional 15 min. Finally, it was washed twice with 1× PBS buffer. Fat body cell images were captured using a Nikon bright-field inverted light microscope (Tokyo, Japan). Day 0 was the control group. We set three repetitions, each with six A. chinensis (three males and three females).

SGC-7901 and BGC-823 cells were grown on coverslips (1 × 10⁵ cells/coverslip), incubated with CHP (0, 20, 30, or 40 μg/mL), and then fixed in ethanol:acetic acid solution (3:1, v/v). The coverslips were mounted on glass slides for the morphometric analysis. Three monolayers per experimental group were photographed. The morphological changes of SGC-7901 and BGC-823 cells were determined using a Nikon bright-field inverted light microscope (Tokyo, Japan) at 400× magnification.