Jsm 6500 tfe sem

MG63 cells were seeded on the nanodots with a different dot diameter and inter-dot distance. After removing the culture medium, the wells were rinsed with Dulbecco’s Phosphate Buffered Saline (DPBS). The cells were fixed with 1.25% gluteraldehyde in PBS at room temperature for 15 minutes, followed by staining with 1% osmium tetraoxide for 30 minutes. Samples were then washed with PBS 3 times, 5 minutes each and finally immersed in 40% alcohol overnight. Dehydration was performed the next day using a series of ethanol concentrations (10 min incubation each in 50%, 60%, 70%, 80%, 90%, 95%, 100% ethanol). The samples were sputter-coated with Platinum and examined by using by JEOL JSM-6500 TFE-SEM at an accelerating voltage of 8 Kiloelectron volts (KeV).

The C6 glioma cells were seeded on the different nanodot surfaces at a density of 5.0 × 10³ cells/cm² for 24, 72, and 120 h of incubation. After removing the culture medium, the surfaces were rinsed three times with PBS. The cells were fixed with 1.25% glutaraldehyde in PBS at room temperature for 20 min, followed by post-fixation in 1% osmium tetroxide for 30 min. Dehydration was performed by 10-min incubation in each of a graded series of ethanol concentrations (40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, and 100%); after which, the samples were air dried. The specimens were sputter-coated with platinum and examined with a JEOL JSM-6500 TFE-SEM at an accelerating voltage of 5 kiloelectron volts (keV). The astrocytic syncytium level of the cells grown on the nanodots was quantified using ImageJ software and compared to the surface area of cells grown on a flat surface. The SEM images of six different substrate fields were measured per sample, and three separate samples were measured for each nanopore surface.

Nanodot arrays were fabricated as previously described [3 (link)]. A 200-nm-thick tantalum nitride (TaN) thin film was sputtered onto a 6-in silicon wafer (Summit-Tech, West Hartford, CT, USA) followed by deposition of 3 µM-thick aluminum onto the top of a TaN layer by thermal coater. Highly uniform nanodot arrays were fabricated from 10 to 200 nm. Nanodot arrays of size smaller than 10 nm could not be fabricated due to technical limitation. Anodization was carried out in 1.8 M sulfuric acid at 5 V, 90 min for the 10 nm nanodot array and in 0.3 M oxalic acid at 25 V, 90 min for the 50 nm. 100 and 200 nm nanodot arrays were fabricated by a two-step anodization method. In the first anodic oxidation step, anodization was carried out in 0.3 M oxalic acid at 40 Volts, 10 min, for 100 nm nanodot array and in 5 % (w/v) H₃PO₄ at 100 V, 5 min, for 200 nm nanodot arrays. The porous alumina was removed by immersion in 5 % (w/v) H₃PO₄ for 70 and 60 min for 100 and 200 nm nanodot arrays, respectively. Second anodization step is repeated in the same way. Porous anodic alumina was formed during the anodic oxidation. The porous alumina was removed by immersing in 5 % (w/v) H₃PO₄ overnight. The dimensions and homogeneity of nanodot arrays were measured and calculated from images taken by JEOL JSM-6500 TFE-SEM. A thin layer of platinum was sputtered onto the structure.