Esem quanta 200 scanning electron microscope

Treated and untreated titanium plates, plated with both MLO-Y4 and 293 cell lines, were observed with the ESEM Quanta-200 scanning electron microscope (Fei Company, Hillsboro, OR, USA) under low vacuum condition and in backscattered mode after gold-palladium sputtering of samples. Samples’ compositions were also analyzed via X-ray microprobe using the ESEM software (version 4.07, Oxford Instruments Analytical, Tubney newlyWoods, Abingdon, Oxon, UK).

Samples (four ossicles) were observed with the ESEM Quanta-200 Scanning Electron Microscope (Fei Company, Hillsboro, OR, USA). The samples were fixed for 20 min in 4% paraformaldehyde in PBS (pH 7.4) and dehydrated through a graded series of alcohol (50%, 75%, 95%, 100%) for 10 min each. The specimens were finally dried overnight and coated with gold-palladium (10 nm thickness) using an Emitech K550 sputter coater (Quorum Technologies, South Stour Avenue, Kent, UK). SEM images were collected under low and high vacuum conditions using a large field detector (LFD) and an Everhart Thornley detector (ETD), respectively. Finally, two selected samples were also analyzed via X-ray microprobe (X-ray energy dispersion spectroscopy, X-EDS) using the ESEM software INCA Suite (version 4.07, Oxford Instruments Analytical, Tubney Woods, Abingdon, Oxon, UK) acquiring five spectra for each SO.