Tecnai g2 spirit bio transmission electron microscope

OMVs were extracted by an electrophoresis and dialysis-based method (ELD) with a 300 kDa cut-off dialysis bag [17 (link)]. Specially, 200 ml of DMEM (Gibco, Shanghai, China) inoculated with the single colony was incubated at 37 ℃ with shaking at 200 rpm for 20–24 h. The culture solution was centrifuged at 10,000×g for 10 min to collect the culture supernatant, which was then filtered through a 0.22 μm filter (Beyotime, Shanghai, China) to remove bacteria and bacterial debris. Next, the OMVs of CRKP was isolated from filtered supernatant as previously described [17 (link)].
The OMVs of CRKP were photographed by transmission electron microscopy (TEM). Five µL of OMV suspension fixed with anhydrous ethanol was dropped onto a 200-mesh copper wire at room temperature for 1 min. Then, the sample was drained with filter paper and negatively dyed with 2% uranium acetate for 1 min. Afterwards, the excess dye was drained with filter paper. Then, the OMVs were observed by a Tecnai G2 Spirit Bio transmission electron microscope (FEI, America). NanoSight NS500 (Thermo Scientific, America) was used for determination of OMVs, and the particle size and concentration of OMVs were analyzed by NTA software.

Expansion of Rhizoctonia cerealis on the wheat leaf sheath was observed with a Nova Nano SEM-450 scanning electron microscope and photographed. The leaf sheath treatment process was conducted basically as described by Xu et al. [95 (link)] and Zhang et al. [96 (link)]. To observe the fungal infection process on wheat leaf sheaths, semi-thin and ultra-thin sections of the leaf sheaths were fixed, dehydrated, infiltrated, and embedded according to Yang et al. [97 (link)], and stained as described by Wang et al. [98 (link)]. Semi-thin cross sections with a thickness of 1 μm were stained with toluidine blue and observed by a microscope with Axio Imager A2. Observation and photography of ultrathin sections were conducted using a TECNAI G2 SPIRIT BIO transmission electron microscope (FEI Company, Hillsboro, OR, USA).

The polysaccharide samples were dissolved in 0.9% NaCl solution (3 mg/mL) for conducting dynamic light scattering (DLS) measurements using a Brookhaven light scattering spectrometer (BI-200SM, Brookhaven, GA, USA) at a wavelength of 532 nm and a 90° angle. The sample solutions were diluted to varying concentrations (0.1–0.75 mg/mL) by using 0.9% NaCl solution and stirred for 24 h after each dilution. The solutions were measured at angles of 50°–140° for static light scattering (SLS). All solutions were purified (filter: 0.22 μm) three times before testing. The obtained data were fitted using Berry plots.
The chain conformation of the polysaccharides was assessed using a Tecnai G2 Spirit Bio transmission electron microscope (TEM; FEI, Portland, OR, USA) at 80 kV. MSP1-1 or MSP1-2 was dissolved in 0.9% NaCl solution to achieve a final concentration of 5 μg/mL. A drop of the polysaccharide solution was deposited on the support carbon film (200 mesh), dried in the air, and stained with 0.2% phosphotungstic acid.