Fei talos l120c

Small pieces of spleen specimens were fixed in glutaraldehyde for 1.5 h and then postfixed in osmium tetroxide (1%). Ultrathin Sects. (70 nm) were placed on copper grids, infused with uranyl acetate and lead citrate, and scoped. Next, Bio-TEM (FEI Talos L120C, Thermo Scientific) was performed to observe the intracellular distribution of the NPs in vivo.

The potential effects of CFS on cell morphology and structure of E. coli ATCC25922 and S. aureus ATCC6538 were determined by SEM and TEM [56 (link),57 (link)]. Briefly, bacterial dilutions (10⁷ CFU/mL) in LB medium were supplemented with 1/2 MIC of CFS and incubated for 10 h at 37 °C by shaking. After centrifugation at 4 °C for 5 min, bacterial cells were collected, washed with PBS (0.1 mM, pH 7.2) three times, fixed overnight with 4% glutaraldehyde, washed with PBS once, and post-fixed in 1% OsO₄ for 30 min. Some post-fixed cells were dehydrated in a graded ethanol series (50%, 70%, 80%, 90%, and 100%), dried by CO₂ for 4 h, coated with gold, and observed under a field emission scanning electron microscope (FEI/Talos L120C, Thermo Fisher Scientific, New York, NY, USA). Other post-fixed cells were dyed with uranium acetate overnight, dehydrated in a graded ethanol series (30%, 50%, 70%, 85%, 95%, and 100%), embedded in Spurr resin, polymerized at 55 °C for 48 h, and observed under a transmission electron microscope (ZEISS, Oberkochen, Germany).

A total of 10 μg PAM trimer and 5 μl (40 mg ml⁻¹) BLP were incubated on a shaking table at 4°C overnight, followed by 5 min centrifugation and resuspension in 1 ml of 1× PBS, three times. BLP (5 μl, 40 mg ml) was obtained following the same protocol as for the control. Further, 10 μl aliquots of BLP‐PAM and BLP were separately applied onto a carbon‐counted 400 Cu mesh grid for 20 min and then blocked for 10 min using 5% BSA in 1× PBS. Next, 10 μg ml⁻¹ VRC01 PBS containing 1% BSA was applied on the grid for 90 min, followed by three washing steps with 1× PBST (1 l PBS and 2 ml Tween‐20). Immunogold goat anti‐human IgG conjugates were applied onto the grid for 30 min, followed by washing with 1× PBST three times. Lastly, the samples were negatively stained with uranyl acetate for 90 s, washed for 2 min with pure water and air‐dried. TEM studies were conducted using a transmission electron microscope (FEI Talos L120C, Thermo Fischer, Waltham, MA, USA) operating at 120 keV.