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Mosquito eggs were pre-fixed with 4% glutaraldehyde for 1 h and then post-fixed in 1% osmium tetroxide for 1 h. Each sample was washed three times with 0.1 M phosphate buffer (pH 7.4). Then, samples were dehydrated for 30 min each with increasing concentrations of ethanol (30%, 50%, 70%, 90%, and 100%) and then placed in 100% acetone for another 30 min. Subsequent critical-point drying and gold coating of particles were performed by the National Taiwan University TechComm. Gold-coated egg samples were analyzed with an FEI Inspect S scanning electron microscope (Thermo Fisher Scientific, Inc.).

Using similar conditions as previously reported [18 (link),19 (link),21 (link),24 (link),25 (link),26 ,27 (link)], scanning electron microscopy (SEM) of the Lactomorphin and Trehalose (as supplied by the manufacturer), SD, and co-SD powders were evaluated. Visual imaging, analysis of particle size, morphology, and surface morphology were achieved by (SEM). Powder samples were attached to aluminum SEM stubs (Ted-Pella, Inc., Redding, CA, USA) using double-sided carbon conductive adhesive Pelco tabs^™ (TedPella, Inc., Redding, CA, USA). Subsequently, the powder sample in the stub was sputter-coated with a 7 nm thin film of gold using Anatech Hummer 6.2 (Union City, CA, USA) system at 20 μA for 90 s under an argon plasma. SEM images of the powder sample were collected using a FEI Inspect S (FEI, Hillsboro, OR, USA). Images were collected using a tungsten source at 30 kV with a working distance of 10–10.4 mm. SEM images were captured at several different magnification levels.

Following the fermentation phase, the BNC underwent treatment with NaOH and distilled water, followed by a drying process through freeze-drying at −80 °C using the 4.5 L FreeZone^® system (Labconco, Kansas City, MO, USA). Subsequently, morphological analysis of the BNC was conducted using an FEI^® Inspect S SEM scanning electron microscope (FEI Company, Hillsboro, OR, USA). During the preparation of the SEM images, the dried BNC was fixed and coated with a thin layer of gold nanoparticles. The experiments were performed at a magnification of 10,000× and an accelerating voltage of 14.5 kV.

The surface microstructures of the CMC/WPI-AF membranes with different CMC:WPI-AF mass ratios were examined using a scanning electron microscope (FEI Inspect S; FEI, USA). In a typical experiment, the CMC/WPI-AF membranes were dehydrated via air-drying at 100 °C for a day. Next, the surface of each membrane was coated with platinum using a metal coating device (VD MSP-1S) under vacuum conditions. Finally, the surface microstructure of each membrane coated with platinum was examined and imaged by means of SEM.