Nova nanosem 230 microscope

Fluorescence live cells images were acquired with Metamorph software using an upright microscope (Nikon eclipse Ni-U) with a 60× water dipping objective (NIR Apo 60X/WD 2.8, Nikon) and an Orca Flash 4.0 camera (Hamamatsu). Fluorophore emission was collected every 3 s. Cells were imaged in a relaxed state and then for 3 min at 5% stretch, and for 3 min during the release of stretch. Fixed cells images were acquired either in the abovementioned acquisition system except for the experiments related to WAVE protein enrichment. In this case, images were acquired in a Zeiss Airyscan microscope (Zeiss LSM880 inverted confocal microscope objective, using Zeiss ZEN2.3 SP1 FP3 [black, version 14.0.24.201] software and a 63×1.46 NA oil immersion objective). Z-stack of single cells were acquired in full Airyscan mode to visualize the PM and the WAVE-C-GFP protein. SEM images were taken using the xTm Microscope Control software in a NOVA NanoSEM 230 microscope (FEI Company) under the high vacuum mode using ET and TL detectors to acquire high and ultra-high resolution images of the cell surface. TEM samples were observed in a Jeol 1010 microscope (Gatan, Japan) equipped with a tungsten cathode in the CCiTUB EM and Cryomicroscopy Units. Images were acquired at 80 kV with a CCD Megaview 1kx1k.

Electrochemical measurements were performed using potentiostat Metrohm^@ µAutolab type III (Eco Chemie, Utrecht, The Netherlands) integrated with screen printed junction cable controlled by NOVA 1.11 software. The SPGE was purchased from DropSens (Oviedo, Spain). The diameter of the disk-shaped working electrode was 4 mm. The working electrode and auxiliary electrode were made of gold, whereas the reference electrode was made of silver. The electrodes were all printed on a ceramic support (L 33 × W 10 × H 0.5 mm). All electrochemical measurements were performed at room temperature. Field emission scanning electron microscopy (FESEM) analysis was carried out using an FEI Nova Nanosem 230 microscope equipped with an energy dispersive X-ray (EDX) system. Raman spectroscopy studies were performed with a WITec Alpha 300R microscope while Fourier transform infrared spectroscopy (FTIR) coupled with attenuated total reflectance analysis were executed using Shimadzu.

Scanning electron microscopy (SEM) investigations were carried out with a high-resolution NOVA NanoSEM 230 microscope (FEI, Eindhoven, Netherlands)) under high vacuum conditions using an accelerator voltage of 5 kV and magnification of 50× to 15,000×. The samples were coated with a gold layer with the ion sputtering method. Two types of microscopic observations were made: surface morphology and cross-section views of the nonwoven fabrics before and after the thermal stabilization process. The transverse dimensions of the fibers in the nonwoven fabric structure were analyzed using the Lucia G software (version 4.8, Laboratory Imaging Prague, Czech Republic) for image analysis. Sixty different fibers from the SEM surface morphology images were selected from randomly chosen places on the nonwoven sample for transverse dimension examination.