Sc7620

Micrographs of the prepared samples were taken by the scanning electron microscope Nova NanoSEM 450 (FEI, Hillsboro, OR, USA) equipped with a high vacuum detector and operated at 5 kV with a spot size of 2.5 A coating with a thin layer of gold/palladium was performed by a sputter coater SC 7620 (Quorum Technologies, Newhaven, East Sussex, UK).

Surface morphology, size, and shape of the MPs fabricated by formulation runs were obtained using a scanning electron microscope (JOEL JCM-5700, USA). The dried MP samples were deposited onto adhesive carbon tabs (Agar Scientific G3357N), which were pre-mounted onto aluminum stubs (Agar Scientific JEOL stubs G306). The samples were gold sputtered for 60 s to attain a thickness of approximately 30 nm (Quorum SC7620). The morphologies of the MP samples were analysed at magnifications of X100, and X500.

To observe microhair changes of Z. macrostachya between the control group and NaCl-treated group, different positions (upper, middle, and lower) of leaf tissue were collected for SEM observation. Leaves of control group and NaCl-treated group were cut into small pieces and fixed in 2.5% glutaraldehyde solution. Samples were stored in 4 °C for 24 h, glutaraldehyde solution was replaced twice during the period of storage. Samples were washed with 0.1 mol/L phosphoric acid buffer for four times, fixed with 1% osmic acid for 1.5 h, then rinsed with deionized water and dehydrated with different concentrations of ethanol (30%, 50%, 70%, 85%, 95% for 15 min for each concentration). Then samples were treated with 100% ethanol twice, 20 min each time. Moreover, the samples were soaked in isoamyl acetate for 30 min and dried at the conventional critical point (E3100, Quorum, UK) [121 (link),122 (link)]. Materials were positioned on the sample stand, and metal spraying (SC7620, Quorum, UK, 2 min) was carried out before observation. SEM was performed on Hitachi TM3030. Energy spectrum was used to examine element content (weight %) of leaf tissue.

Wood samples were cut in to 0.5 × 0.5 × 3 cm³ block. The blocks were sectioned with a sliding microtome at 120 µm thickness. Sections were dried in a hot air oven (80° C) for 48 h and then attached on a stub with a double-sided carbon tape. Samples were coated with gold particles using a sputter coater machine (SC7620; Quorum, England) and observed under a scanning electron microscope (JSM 5600 LV; JEOL, Tokyo, Japan).

The morphology of the fracture surfaces of composites from the impact tests, was observed by field emission scanning electron microscopy (FESEM) using a ZEISS ULTRA 55 from Oxford Instruments (Abingdon, UK) microscope. Before placing the samples in the vacuum chamber, the samples were sputtered with a gold-palladium alloy in an EMITECH sputter coating model SC7620 from Quorum Technologies, Ltd. (East Sussex, UK) applying an acceleration voltage of 2 kV.

To detect the microstructure of GelMA hydrogel and the distribution of S. boulardii in bioactive materials, the samples were pre-cooled at −80 °C for 1 h and then freeze-dried by a freeze dryer (SCIENTZ-10N, Xinzhi, Ningbo, China) at −60 °C for 24 h. After being sprayed with conductive material by Quorum SC7620, the samples were observed using a scanning electron microscope (TescanmtraLms, Brno, Czech Republic). To improve the visualization of the pore structure, the GelMA solution was labeled with rhodamine B. After photo-crosslinking, GelMA hydrogel containing rhodamine B was soaked thoroughly. GelMA hydrogel was washed and observed using a fluorescence microscope. Likewise, the fluorescein labeling method was used to observe the activity of S. boulardii bioactive materials. S. boulardii was stained by Calcofluor White (CFW; final concentration: 10 mg/mL) for 10 min [28 (link)]. After being washed several times, S. boulardii bioactive materials were observed.