Nanowizard 4a nanoscience

SEM microscopy was performed to investigate the microstructure of tendon tissue, pattern, and topography of cell replica and tissue replica and cell morphology on sheets. For cell fixation, 2.5% glutaraldehyde was added to all samples so as to cover all the sheets. All samples were dehydrated using ethanol with increasing concentration (serial dilution 50-100%). After drying, the specimens were coated with a thin layer of gold and then were viewed under a TESCAN VEGA3 SEM at an accelerating voltage of 30 kV. In order to investigate the topography of cell replica, AFM (NanoWizard 4a NanoScience, JPK Instruments AG, Berlin, Germany) contact A c c e p t e d M a n u s c r i p t mode imaging was performed. A rectangular cantilever (HQ:NSC18/Al BS, MikroMasch, Bulgaria) was used with spring constant of 2.8 N/m and a conic tip of 8 nm radius. Using a scan rate of 0.05 Hz and setpoint force of 0.5 nN, surface areas up to 100 2 µm were scanned. Image analysis was done using standard software of the instrument (JPKSPM Data Processing).

The topography of the fabricated substrates and the cells morphology cultured on the imprinted substrates after 14 days were visualized by SEM (Vega2 Tescan, Czech). Before imaging, cells were fixed in 4% glutaraldehyde solution for 45 minutes followed by dehydration with graded dilutions of ethanol (10-100% v/v). Additionally, samples were coated in gold, and SEM was carried out at various magnifications.
The morphology and topography of the substrates and cultured cells on samples were also analyzed using AFM contact mode imaging (NanoWizard 4a NanoScience, JPK Instruments AG, Germany). The instrument software (JPKSPM Data Processing) was used to analyze and obtain the cross-section height profiles of the images.