Tcs sp8 inverted confocal laser microscope

For immunocytochemical analysis, BMEC were seeded onto glass coverslips in 6-well plates at a density of 1 × 10⁵/well, and allowed to attach for 24 h. After washing with PBS, cells were fixed in 4% (wt/vol) paraformaldehyde for 15 min on ice, then rinsed 3 times with TBS-T (20 mM Tris-base, 140 mM NaCl, 0.1% [vol/vol] Tween-20) and blocked with 5% (wt/vol) BSA in TBS-T for 1.5 h at 37 °C. Subsequently, cells were incubated with mouse monoclonal anti-CK18 antibody at 4 °C overnight, washed with PBS, incubated with FITC-conjugated goat anti-mouse IgG at 37 °C in the dark for 1 h, washed with TBS-T, followed by incubation with DAPI in the dark for 20 min. After mounting of the cells in Antifade Mounting Medium, images were obtained with a Leica TCS SP8 inverted confocal laser microscope and Leica software (Wetzlar, Germany).

To investigate the possible effects of CQDs on B. xylophilus, three samples of the nematodes immersed in CQDs solution (4 mg mL⁻¹) was considered as the G treatments. The treated nematodes were collected for confocal imaging. Worms were washed three times with sterilized distilled water and fixed with 10% formaldehyde. Subsequently, worms mounted on slides following standard procedures. Leica TCS-SP8 inverted confocal laser microscope was used. Images were viewed and processed with Leica image analysis software. Twenty nematodes were examined, and three replicates were performed.