Xl 30 esem feg scanning electron microscope

Collagenase Type II enzyme (1 mg/mL; Invitrogen Corporation, USA), Hank’s Balanced Salt Solution 1X, 14025, Gibco (HBSS), RPMI–1640 (Sigma-Aldrich GmbH, Germany), DMEM (1000 mg glucose/L, Sigma Chemical St. Louis, USA) Penicillin-Streptomycin, fetal bovine serum (Sigma Chemical St. Louis, USA), Osteoblast Stimulator MesenCult-XF kit (MSC Basal Medium human-05431, Osteogenic Stimulatory Supplement human-05435, and β-Glycerophosphate human-05436), which is used as osteogenic progenitor, were obtained from Stemcell Technology, USA (≠9966/05434).
The laminar flow cabinet (Air Flow-NUVE/NF–800 R) and the incubator (NUVE) were obtained from Akyurt, Ankara, Turkey. Primary cell culture consumables (flask, 6-well plate, pipette, tube, and scraper) were obtained from Sarstedt, Greigner. A CKX41 invert microscope, and for ELISA measurements, a Mindray MR 96 A were used (PRC). A Quanta 250 FEG (Fei Company, Hillsboro, Oregon, USA) environmental scanning electron microscope (ESEM) was used. For porosity assessment of the plates and surface morphology characterizations, a FEI-Philips XL30 ESEM-FEG scanning electron microscope and gold-plated Polaron SC7640 Sputter Coater were used. The antibodies used for immune phenotyping were BD brand and they were analyzed using a BD FACS-Calibur flow cytometer.

Scanning electron microscopy (SEM) was used to analyze the microstructure of all the groups. The SEM was evaluated by the method of Palanisamy et al. (2018). The refrigerated samples were taken out and cut into 10 × 10 mm, then dried immediately to the full. Then, the sample surfaces were sputter coated with gold and images were taken by XL‐30 ESEM FEG scanning electron microscope (FEI, Shanghai, China).

The compressive mechanical test was performed on the 3D PEI scaffolds (20 ∗ 12 ∗ 12 mm) and PEI rods (12.7 mm ∗ 10 mm) via the electronic universal testing system (Instron 5869, USA) with 50 kN load cells and the crosshead speed set as 1 mm/min. The compressive strength, compressive modulus, and stress-strain curve were obtained from the load recorded. The structure and the surface of the 3D PEI scaffold were observed via FE-SEM (XL-30 ESEM FEG Scanning Electron Microscope, FEI Company). The samples were sputtered with Au before SEM observation.

A subset of seventeen samples also underwent ultrastructural characterization by scanning electron microscopy (SEM). SEM analysis was conducted by using both environmental (E-SEM) and conventional high-vacuum (HV-SEM) operative modes. The E-SEM mode allowed imaging of fully hydrated samples without the need for conductive coating, thus informing the native status of the sample. HV-SEM was applied to obtain high-resolution images of micromorphologies at the membrane surface. Differently from E-SEM, HV-SEM required complete dehydration and metallic coating for guaranteeing sample conductivity. HV-SEM preparation was obtained by dehydration in ascending hydroalcoholic solutions, drying in a laminar flow cabinet, mounting on a sample holder, and sputter-coating with gold. SEM imaging in both E-SEM and HV-SEM modes was performed with a XL30 ESEM FEG scanning electron microscope (FEI, Philips, The Netherlands). High-resolution micrographs were obtained with a magnification ranging from 500 to 2000 time in the E-SEM mode and up to 10000 times in the HV-SEM mode. Details of surface morphology and any eventual cell-like structure were imaged and stored.

High-resolution transmission electron microscopy (HR-TEM) pictures were collected at 100 kV using a TECNAI G2 microscope (Thermo Fisher, Waltham, MA, USA). The image of scanning electron micrographs (SEM) was measured by a XL-30ESEM FEG scanning electron microscope (FEI, Hillsboro, OR, USA). FTIR spectra were conducted with a VERTEX 70 FT-IR spectrometer (Bruker, Bremen, Germany). The X-ray photoelectron spectroscopy (XPS) spectra were acquired using an ESCALAB 250Xi spectrometer (Thermo Fisher, Waltham, MA, USA). A Rigaku Minister apparatus was used to generate the X-ray diffraction (XRD) patterns (Tokyo, Japan). The UV absorption spectra were performed via a Hitachi UV2450 spectrophotometer (Tokyo, Japan). The fluorescence spectra were obtained using an F97Pro FL spectrophotometer coupled with a 1.0-cm quartz cell (Lengguang Technology, Shanghai, China). Additionally, the fluorescent, phosphorescent lifetime, and emission spectrum were analyzed at room temperature using an FLS-1000 fluorescence spectrophotometer (Edinburgh, UK).