Cm120 transmission electron microscope

The size and size distribution of DCM-[PTX/DiD] NPs were measured by dynamic light scattering instruments (DLS, Microtrac, USA). The micelles concentration was at 1.0 mg/mL and PTX concentration at a dose of 0.1 mg/mL. The morphology images of NPs were measured under CM-120 transmission electron microscope (TEM, Philips, Holland). The data of NPs was presented as mean + standard deviation. The stability study of DCMs loaded DiD was monitored the change of particle size with or without sodium dodecyl sulfate (SDS) and GSH, which has been reported effectively induced the disintegration of polymeric micelles [57 ]. The particle size of PTX-loaded DCMs was incubated with PBS solution with or without 10% FBS at body temperature (37 °C) by DLS.

After anesthesia, rats at P1 and P7 underwent cardiac perfusion with ice-cold 2.5% glutaraldehyde (no. G5882, Sigma, USA). The cochleas were dissected and fixed with 2.5% glutaraldehyde for 24 h and decalculated in 10% EDTA. Subsequently, the cochleas were fixed with 1% osmic acid for 2 h at room temperature, rinsed with 0.1 M PBS for 3 times, and dehydrated with ethanol and acetone. The cochleas were immersed in Epon 812. Ultrathin sections (60-70 nm) of cochleas were prepared. Then, the sections were stained with alkaline lead citrate and uranyl acetate and observed under a Philips CM-120 transmission electron microscope (Amsterdam, Philips, The Netherlands).

For negative contrast EM, liver tissue was homogenised in Phosphate Buffered Saline (PBS) 20% w/v and centrifuged at 17,000 x g for 5 minutes. The supernatant was adsorbed onto carbon-coated formvar copper grids and stained with nanoW (Nanoprobes, NY, USA) for 1 min. Cell culture media from virus isolation attempts, clarified by centrifugation (as above), were similarly prepared.
For thin section EM, glutaraldehyde fixed liver tissue or cell pellets from virus isolation attempts, were postfixed with 1% osmium tetroxide for 1 hour and embedded in Spurr’s resin (ProSciTech, QLD, Aus) in routine fashion. Ultrathin sections were stained with saturated uranyl acetate in 50% ethanol followed by lead citrate. All prepared grids were examined using a Philips CM120 transmission electron microscope at 120kV.

The samples Ag@MOF, GO, and Ag@MOF–GO as well as their composite hydrogels were recorded on an IRAffinity-1 spectrometer (Shimadzu, Kyoto, Japan) in the range 4000–400 cm⁻¹ with a scanning rate of 2 cm⁻¹. The morphology of Ag@MOF, GO, and Ag@MOF–GO as well as their composite hydrogels were obtained by a JSM-6700F scanning electron microscope (SEM) (JEOL, Tokyo, Japan). Transmission Electron Microscopy (TEM) images were investigated by a Philips CM 120 transmission electron microscope. X-ray diffraction spectra were obtained by an X-Ray diffractometer (ULTIMALV, Japan Science Corporation, Tokyo, Japan). The scanning diffraction angle was between 5 and 50°, and the current was 20 mA. UV spectroscopy was investigated by a Shimadzu UV-2550 spectrometer in the scanning mode of 200–800 nm.

8 × 10⁵A. castellanii were seeded into 6-well plates, infected with L. pneumophila (MOI 20) in minimal medium and incubated at 37°C. After 2 h the medium was replaced with PYG. 8, 16, and 24 h post infection cells were harvested, pelleted by centrifugation (10,000 g, 2 min), resuspended in fixation buffer (2.5% glutaraldehyde in 0.1 M sodium cacodylate) and incubated for 2 h at RT. Cells were then pelleted by centrifugation and washed twice with 0.1 M sodium cacodylate before post-fixation in 1% osmium tetroxide for 2 h at RT. Cell pellets were then washed in dH₂O and left overnight at 4°C in 0.3% uranyl acetate. Samples were rinsed in dH₂O before dehydration in a graded series of acetone and then infiltrated and embedded with EPON resin. Sections (70–80 nm thick) were cut, stained with uranyl acetate and lead citrate before viewing under a Phillips CM120 transmission electron microscope at 120 Kv.