Cf400 cu

Purified IAV samples were pH-adjusted and neutralized using 10 × pH buffers as described above. All samples were pH exposed for 30 s at ambient room temperature prior to neutralization. Neutralized samples were then diluted to 10¹⁰ PFU/mL equivalent in 1 × pH 7.0 buffer and fixed with 0.06% paraformaldehyde for 4 d at 4°C prior to use in TEM. Five microliters of each fixed sample was deposited onto carbon-coated copper grids (Electron Microscopy Sciences, CF400-Cu). Grids were UV-glow discharged immediately prior to sample addition, and the sample was left to adsorb for 2 min at room temperature. Excess solution was blotted using Whatman filter paper, and grids were washed in a droplet of miliQ water. Samples were then negatively stained for 1 min with 5 µL of 2% uranyl acetate (Electron Microscopy Sciences, 541–09-3). Excess stain was blotted, and grids were allowed to air dry for 5 min. Grids were imaged using the Tecnai Spirit TEM (spot size: 2, emission: 4.41 uA, and accelerating voltage: 80 kV).

The EV separated from CTE and control brain tissue were analyzed by TEM. The EV sample (5µl) was adsorbed for 1 min to a carbon-coated mesh grid (# CF400-CU, Electron Microscopy Sciences) that had been made hydrophilic by a 20-s exposure to a glow discharge (25 mA). Excess liquid was removed with a filter paper (#1 Whatman). The grid was then floated briefly on a drop of water (to wash away phosphate or salt), blotted on a filter paper, and then stained with 0.75% uranyl formate (# 22451 Electron Microscopy Sciences) for 30 s. After removing the excess uranyl formate, the grids were examined, and random fields were photographed using a JEOL 1200EX TEM with an AMT 2k CCD camera.

Morphology of freshly prepared original samples and redispersed samples was observed under a transmission electron microscope (TEM). Firstly, diluted samples were loaded onto a plasma-cleaned carbon TEM grid (CF400-CU, Electron Microscopy Science, Hatfield, PA, USA) and stained with 0.5% uranyl acetate stain solution. Then, the morphology was captured by a TEM (FEI, Tecnai 12 G2, Spirit, BioTWIN, Eindhoven, Netherlands) equipped with a CCD camera (AMT 2k XR40). For the spray-dried powder samples, the morphology was observed by a scanning electron microscope (SEM, JSM-6330F, JEOL Ltd., Tokyo, Japan), by mounting the powder on a double-adhesive carbon tape pre-affixed on a specimen stub.

The scanning electron microscope (SEM, JSM-7600TFE, FEG-SEM, Calgary, AB, Canada) at an operational voltage of 2 kV was used to study the morphology of electrospun fibers. Fiber diameters were calculated using Image-Pro Plus® software by taking an average of about 300 fibers. To confirm the presence of graphene sheets in the nanofibers of PANI/PMMA/Am-rGO, transmission electron microscopy (TEM, JEOL, JEM 2100F, JEOL, Pleasanton, CA, USA) was used. For TEM observation, fibers were directly deposited onto a TEM copper grid with supporting carbon film (CF400-Cu, Electron Microscopy Sciences). Thermogravimetric analysis (TGA) was conducted under nitrogen atmosphere using Q5000 TGA (TA Instruments, New Castle, DE, USA) in the temperature range of 20–900 °C, with a heating ramp of 10 °C·min⁻¹.

The pH-MUC1-Pt NPs’ size and zeta potential were analyzed by dynamic light scattering (DLS) and ZetaPals (Brookhaven Instrument Corporation). The concentrated NPs solution was diluted in distilled water to obtain a concentration of 0.500 mg PLGA/mL. The obtained solution was measured for the hydrodynamic mean diameter and size distribution. Each measurement was performed in triplicate. The shape, surface morphology, and size of the NPs were analyzed by TEM. The NP samples were mixed with the acetate buffer (0.125 M CH₃COONH₄, 0.6 mM (NH₄)₂CO₃ and 0.26 mM tetrasodium EDTA at pH 7.4). A total of 10 μL of the sample was negatively stained with 10 μL of 2% (w/v) phosphotungstic acid. A droplet of the NPs was placed on a carbon-coated copper grid (Electron Microscopy Sciences, CF400-Cu, carbon film, 400 mesh), forming a thin liquid film.