Tm3000 benchtop sem

ECM were fixed with 4% paraformaldehyde in phosphate buffer (PB) for 5 h, postfixed in 1% osmium tetroxide in PB for 1 h, dehydrated in ascending degree of ethanol (70–80–95–100%), dried for 30 min in xylene and embedded with paraffin after evaporation of xylene and serially cut as 10 μm thick sections using a Leica microtome. Sections were first deparaffinized in xylene for 2 h and then hydrated in descending degree of ethanol (100–95–80–70%) and H₂O. Ultrathin sections were collected on glass coverslips, mounted on aluminum stubs and sputter-coated 3 min with gold. Morphology and ultrastructure were investigated by SEM on a TM3000 Benchtop SEM (Hitachi, Pleasanton, CA, USA) instrument operating at 15 kV acceleration voltage [30 (link)]. Areas of matrix or empty spaces were calculated with a tool of the ImageJ software (version 2.0, National Institute of Health, Bethesda, MD, USA) and reported as percentage area on the whole image analyzed. Results are also reported as perimeter/area ratio of empty spaces. Repopulated GelMA scaffolds were analyzed by SEM as above, after fixation without embedding.

The morphology and structure of the samples were assessed using scanning electron microscopy (SEM) performed on a Hitachi TM 3000 Benchtop SEM instrument (Tokyo, Japan) operating at 15 kV acceleration voltage. Observations were carried out on fragile fractures (in liquid nitrogen) of samples lyophilized and sputter-coated with gold.
Observations at higher magnification were carried out with a commercial AFM (NTMDT) model NTEGRA in tapping mode. For AFM measurements, the oxidized PS sample was fixed on a glass slide with a double tape.