Em scd050

Adult worms collected by perfusion were immediately transferred to supplemented RPMI medium; parasites were distributed in 6-well plates (adults: 10 paired worm couples per well) with medium. The worms were kept in culture (oven at 37°C and 5% CO₂) for 2 h for adaptation, and then NJ series compounds or the equivalent amount of 0.1% DMSO vehicle were added.
Ultrastructural analysis was performed with scanning electron microscopy. Adult worms incubated at different concentrations of NJ series compounds or with 0.1% DMSO vehicle for 1 and 2 days were fixed with modified Karnovsky reagent (1% paraformaldehyde, 2.5% glutaraldehyde, 1 mM calcium chloride in 0.1 M sodium cacodylate buffer, pH 7.4) and after the fixing stage the material was washed with sodium cacodylate buffer (0.1 mol / L, pH 7.2) and post-fixed with 1% osmium tetroxide (w / v) for 1 h.
Samples were dehydrated with increasing concentrations of ethanol and then dried with liquid CO₂ in a critical-point dryer machine (model Leica EM CPD030, Leica Microsystems, Illinois, USA). Treated specimens were mounted on aluminum microscopy stubs and coated with gold particles using an ion-sputtering apparatus (model Leica EM SCD050, Leica Microsystems, Illinois, USA) [34 (link)]. Specimens were then observed and photographed using an electron microscope (FEI QUANTA 250, Thermo Fisher Scientific, Oregon, USA).

Candida albicans B16 cells were treated with E15K at 5×C_L at 30 °C for 6 h and were fixed with 2.5% glutaraldehyde for 1 h, followed by washing three times with PBS. Cells were dehydrated with a series of graded ethanol solution and then dried by a critical point dryer (Leica EM CPD300, Austria) before being mounted on carbon tape, sputtered with gold coating (Leica EM SCD050, Austria). Images were visualized by a SEM (FEI QUANTA 450, Czekh).

Micrographs of freeze-dried samples were measured on a Zeiss Supra 55 VP. Before the measurement, the samples were sputtered with a 10 nm-thick gold layer (Leica EM SCD050, Wetzlar, Germany).

A 10 × 3 × 1 mm piece of nickel, titanium or nitinol with BEAS-2B cells cultured on its surface were fixed for Scanning Electron Microscopy (SEM) in 2% (v/v) glutaraldehyde in 0.1 M cacodylate buffer, and stored until usage at 4 °C. Next, the samples were incubated for 1 h in 1% (v/v) osmium tetroxide (OsO₄) in 0.1 M cacodylate buffer and afterwards the metals were washed three times with Milli-Q ultrapure water. The samples were dehydrated in 30%, 50%, and 70% ethanol, 10 min each, followed by four times dehydration in 100% ethanol for 10, 20, and two times 30 min respectively. Lastly, the samples were dried by incubating for 10 min in 1:1 100% ethanol/tetramethyl silane (Acros, Geel, Belgium) and for 15 min in pure tetramethyl silane followed by air-drying.
After sample preparation, the pieces of metal were placed on a carbon holder and subsequently sputter coated (Leica EM SCD050 and QSG100, Wetzlar, Germany) with 4 nm palladium/gold.
Samples were imaged with a Zeiss Supra 55 SEM (Oberkochen, Germany) using a secondary electron detector at an acceleration tension of 3.0 kilovolt (kV) with 30 µm aperture at 7.2 mm working distance.

Specimens were examined with a Nikon SMZ 1500 stereomicroscope fitted with a 10 mm ocular grid having 100 divisions. Some ethanol-preserved specimens of parasitoid wasps were critical-point dried with a Leica EM CPD300 automated critical point dryer for morphological studies. Several critical-point dried specimens were dissected into head, mesosoma, and metasoma for scanning electron microscopy (SEM), which were subsequently sputter-coated with gold using a Leica EM SCD050 super cool sputter coater. Micrographs were produced using an FEI Quanta 450 environmental scanning electron microscope. Habitus pictures were taken with a Nikon D7000 digital camera connected to the Nikon SMZ 1500 stereomicroscope and then stacked in Helicon Focus software to generate single highly focused images. All images were processed with Adobe Photoshop CC 2019.
Morphological terminology follows Gibson [12 ]. Abbreviations used for morphological characters are as follows: F1–F3, funiculars 1–3; C1–C3, clavomeres 1–3; MLM, midlobe of mesoscutum; Gtn, gastral tergite number; POL, the shortest distance between the posterior ocelli; OOL, the shortest distance between an eye and posterior ocellus.

After fracture resistance testing, representative crowns from each group were observed by using the TESCAN scanning electron microscopy (SEM) (Mira 3XMU, Kohoutovice, Czech Republic), to check the fractured surface condition and highlight any differences between the groups (if present). Sputter coating of the crown was done with gold (Leica EM SCD050, Wetzlar, Germany) to a thickness of approximately 10 µm prior to imaging.