Scd 050

The changes during the drying were microstructurally examined using Scanning Electron Microscope (a JEOL model JSM 5800LV) at 5 kV. The samples were coated with gold in a Sputter Coater (BALZERS, model SCD050).

For light microscopy (LM), INCs were mounted onto glass microscope slides, suspended in water, sealed with coverslips, and then viewed under an Axiophot light microscope (Carl Zeiss, Jena, Germany) operating in brightfield mode using the objective of 20× magnification. Representative light micrographs of cells were captured using a Leica DFC320 camera equipped with the Leica software application suite LAS V3.8 (Leica Microsystems, Wetzlar, Germany). Digesta samples taken after 0 and 120 min of the small intestinal digestion were stained with 2% (w/v) Lugol’s iodine solution and visualised under LM for detecting the presence of starch.
For scanning electron microscopy (SEM), INCs were directly mounted on double-sided adhesive tapes on aluminium stubs, sputter coated with gold (SCD 050, Balzers, Liechtenstein), and viewed under a scanning electron microscope (FEI Quanta 200 FEI Electron Optics, Eindhoven, the Netherlands). Representative electron micrographs of cell samples were captured with accelerating voltage of 25 kV and using the xT microscope software version 3.0.7 (FEI Quanta, Eindhoven, the Netherlands).

Non-galled leaves and rolling galls fixed in FAA (Johansen, 1940 ) were dehydrated in an ethanolic series (Johansen, 1940 ), critical point dried, mounted on stubs, and covered with 15 nm of gold (Balzers SCD 050) (O’Brien and McCully, 1981 ). The samples were observed in a scanning electron microscope (JEOL JSM - 6360LV) in the Center of Microscopy at the Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais state, Brazil¹.

The microstructure of the gelatinized starch was observed using a scanning electron microscope (TM-1000, Hitachi, Japan). The starch powder was dispersed and uniformly placed on the carrier platform before conductive coating in a sputter coater (SCD 050, Balzers, Liechtenstein) was performed, and then the sample was then firmly fixed by a rubber ear syringe. The images were taken under an acceleration voltage of 2 kV and were photographed at 3000× magnification according to a previous report [66 (link)].

Scanning electron microscopy analysis of the injection molded samples was performed with a field emission SEM (FE-SEM, Merlin^®, Carl Zeiss GmbH, Jena, Germany) using 2 kV acceleration voltage. Prior to imaging the cross-sections, the samples were freeze-fractured under liquid nitrogen and sputter-coated (SCD 050, Balzers AG, Balzers, Liechtenstein) with a thin gold layer.

The scanning electron microscopy (SEM) (S-4800, Hitachi, Tokyo, Japan) was used to evaluate the morphology of the cells attached to the zirconia disks. In this study, 5×10⁴ or 1×10⁴ cells were seeded onto disks and incubated for 3 or 24 h, respectively, to observe the cell attachment and spreading. For conducting the SEM examination, specimens were fixed with 4% formaldehyde for 2 h at room temperature and rinsed three times in deionized water for 15 min; then, each specimen was dehydrated in an ascending ethanol series (ranging from 30 to 100% ethanol), three times for 10 min at 4°C. Finally these samples were sputter coated with gold palladium for 60 s at 60 mA (SCD050, Balzers, Liechtenstein).