Fei quanta 650 feg sem

The morphologies of the Saos-2 cells seeded at 2.5 × 10⁴ cells/well at passage 27 onto ZrO₂ and Y-TZP discs with rough (Ra = 1 µm) and mirror-polished (Ra = 0.04µm) surface conditions during 96 h were observed by SEM. The cells were fixed with 5% glutaraldehyde (Sigma-Aldrich^®, Lot:#SLCC3121, St. Louis, MO, USA) at 4 °C for 30 min, rinsed 3 times with phosphate buffer (Sigma-Aldrich^®, Lot:#SLBZ5107, St. Louis, MO, USA), and then resuspended in 1% osmium (Sigma-Aldrich^®, CAS:20816-12-0, St. Louis, MO, USA) for 30 min. The samples were then dehydrated by a concentration gradient of ethanol in distilled water (35%, 50%, 75%, 95%, 95%, 100%, 100% (v/v)) (Sigma-Aldrich^®, St. Louis, MO, USA) for 15 min each time. Then, the samples were chemically dehydrated using hexamethyldisilane (Sigma-Aldrich^®, Lot:#SHBJ6186, St. Louis, MO, USA) overnight in a vacuum desiccator (12 h). After complete drying, the samples were metalized for 10 min by gold sputtering to obtain a gold layer of ~15 nm (Safematic Compact Coating unit-010, Institut Jean Lamour, Nancy, France) and observed with the FEI Quanta 650 FEG™ SEM (Institut Jean Lamour Nancy, France).

The morphology of SaOs-2 cells, seeded at 2 × 10⁴ cells/well was observed after a 72 h incubation period by SEM. The cells were fixed with 0.5% glutaraldehyde at 4 °C for 30 min, rinsed 3 times with phosphate buffer and then resuspended in 1% osmium for 30 min. The samples were then dehydrated for 15 min with an ascending ethanol gradient, 35%, 50%, 75%, 95%, 95%, 100% (v/v). Next, the samples were chemically dehydrated using hexamethyldisilane overnight in a vacuum desiccator. After complete drying, the samples were metallized for 5 min by gold sputtering to obtain a gold layer of ~15 nm and observed with the FEI Quanta 650 FEG™ SEM (FEI company, Hillsboro, OR, USA)).

We performed high-resolution SEM imaging of fly body parts (head, wings, and thorax). Fly samples were affixed to SEM aluminum stubs using conductive adhesive carbon tape or carbon glue. To enhance electrical conductivity and imaging quality, the samples were coated with a 10-nm layer of gold using a Quoram 150T ES magnetron sputter coater (Quoram Technology) with argon gas. Imaging was performed using a FEI Quanta650FEG SEM (Thermo Scientific, United States of America) operating at 5 kV with a Secondary Electron ETD detector.