Su1510 sem

The 4% (wt/vol) VCL and S‐VCL‐S proteins solution were mixed with 0.1% (wt/vol) H₂O₂. The hydrogels were then lyophilized with a FreeZone Plus 6 L cascade console freeze‐dry system (Labconco, Kansas City, MO, USA). The lyophilized hydrogels were sputter‐coated with a thin layer of a palladium/gold alloy to improve the surface conductivity for SEM. Images of the morphology and microstructure of the hydrogels were acquired using a Hitachi SU1510 SEM (Tokyo, Japan) with an acceleration voltage of 5 kV.

Different concentrations of gelatin solutions were freeze-dried using a freeze-drying system. A thin layer of palladium/gold alloy was placed on the lyophilized hydrogel to improve the surface conductivity for SEM. Morphological and microstructural images of the gelatin were acquired using a Hitachi SU1510 SEM at an accelerating voltage of 5 kV.

FTIR experiments were performed using an FTIR spectrophotometer (Nicolet-460, Thermo Fisher Scientific, Waltham, MA, USA). The complexes obtained after freeze-drying were milled and pressed with potassium bromide and polymer in a ratio of 200:1. Transmittance values between 4000 cm⁻¹ and 400 cm⁻¹ were measured to determine the functional relationship between the different compounds.
XRD analyses of the samples were performed using a smart X-ray diffractometer (SmartLab 3KW, Rigaku Corporation, Tokyo, Japan), according to Dong et al. [20 (link)], with slight modifications. The prepared powder was flattened on a glass plate for X-ray diffraction analysis, and the scanning range was 2θ from 5o to 50°, with a step size of 5 °/min/scan rate.
Scanning electron microscopy (SEM) of the samples was conducted using a SU1510 SEM (Hitachi, Tokyo, Japan). The freeze-dried powder was adhered to a double-sided adhesive tape, coated with gold, and observed using a scanning electron microscope at a magnification of 500×.
An FEI Tecnai 12 (FEI Company, Hillsboro, NH, USA) was used for transmission electron microscopy (TEM). A drop of the complex coacervate solution was placed on a 400-mesh copper grid. The grids were then dried overnight at 40 °C, and the morphology of the samples was observed using an electron microscope with a single-tilt sample rack at an accelerating voltage of 100 kV.

All treated samples were dried using freeze dryer (LABCONCO) and stored at −80 °C. The dried samples were used for observation for scanning electronic microscope (SEM) by Hitachi SU1510 SEM at, 500X, 1000X and 5000X magnification at BSE mode.

Petri dish microcultures of the strain ENCB-J15 were inoculated, grown in squares of GAE solid medium covered by a circular coverslip, and incubated at 28 °C for 10 days. The coverslip with a culture sample was placed on a 12-well microplate and fixed with 2% glutaraldehyde for 2 h, dehydrated with 10, 20, 30, 40, 50, 60, 70, 80, and 90% ethanol each for 10 min for each dissolution, and finally dehydrated with absolute alcohol for 20 min [27 (link)]. The sample was critically dried with CO₂ using the Emitech K850 Critical Point Dryer to remove all moisture and subsequently mounted on aluminum sample holders with adhesive carbon tape. The sample was coated with a layer of gold at 20 mA for 2 min using Quorum Q15OR-ES equipment. Finally, the sample was observed in a Hitachi SU1510 SEM at 10–15 kW [28 (link)].