Jsm 6300f

SEM measurements were performed with a field emission microscope JSM-6300F (JEOL, Tokyo, Japan). The energy of the exciting electrons was mostly 5 keV. Beside the detector for secondary electrons (SEI), Everhart-Thornley type, the system is equipped with different detector types (semiconductor and YAG type) for backscattered electrons. The sample preparation was done by the deposition of 10 µL droplets of the particle dispersion on polished amorphous carbon substrates (sigradur) and dried in air. Because of the low atomic number of carbon this strategy helps to increase the imaging contrast.

The corneoscleral buttons were then dissected and the flap gently cut off. The specimens were fixed in formaldehyde 10% and processed for scanning electron microscopy (SEM). Briefly, washing in BSS for 5 minutes was followed by dehydration in ascending concentrations of ethanol (25%, 50%, 75% and 100%, ten minutes each), followed by infiltration with three changes of hexamethyldisilazane (HMDS) each lasting for 10 minutes. The samples were allowed to slowly air-dry overnight. They were then coated with 20 nm of gold by resistive thermal evaporation and examined (JEOL JSM-6300F, Tokyo, Japan) at 5 kV and 10 mm working distance. SEM photos were taken at 1000× magnification, at three different locations within the ablated bed.
Smoothness of the ablated bed was then evaluated based on the texture analysis of the SEM images [2] (link). Image texture, defined as variations in the pixel intensities (variations in the gray-levels), was analyzed using the Haralick texture contrast parameter [25] . The usefulness of this parameter has been well documented in biomedical sciences [26] (link), and more specifically, for the quantification of corneal stromal surface roughness as observed on a SEM image [27] (link). The Haralick contrast parameter was computed for each SEM image (Matlab R2009b version). Haralick contrast values increase with roughness.

The freeze-dried kafirin microparticle samples were mounted onto carbon tabs and coated with 10 nm of platinum using a sputter coater (Baltec MED 020, Leica Microsystems, Wetzlar, Germany). The samples were then viewed at 5 kV and a working distance of 8 mm using a field emission scanning electron microscope JSM 6300F (JEOL, Tokyo, Japan) [4 (link),23 (link),25 (link)].

A scanning electron microscope (Model JEOL JSM-6300 F, Tokyo, Japan; 2–5 kV with Auto Fine Coater, JEOL-JFC-1600E Ion Sputtering Device) was used to study modified invertase-onion membranes. During SEM analysis, onion membrane(s) were mounted on stubs and coated with Au/Pd. SEM micrographs of both the invertase immobilized and those that were blank were taken at various magnifications.

SEM measurements were performed with a field emission microscope JSM-6300F (JEOL, Tokyo, Japan) and FEI Helios NanoLab G3 UC (ThermoFisher, Nederland). The energy of the exciting electrons was mostly 5 keV. In order to enhance the surface sensitivity and in this manner the topographical impression some of the micrographs were taken at a stage tilt of 45°. Besides the detector for secondary electrons (SEI), Everhart-Thornley type, the system is equipped with different detector types (semiconductor and YAG type) for backscattered electrons.