Evo 60 ep sem

Scanning electron microscope (SEM) images of the clusteroids were taken in order to reveal their morphology. Samples were prepared for SEM images after fixing the cell clusteroids to avoid them to break apart during the evaporation of the aqueous solution. Of the HaCaT cell clusteroids in media, 1 mL was deposited on dry Aclar™ sheets (Agar Scientific Ltd., Essex, UK) or poly-lysine coated glass coverslips and treated with 2 wt% glutaraldehyde for 2 h. This was followed by washing with cacodylate buffer, rinsing with serial ethanol-water solutions of increasing ethanol concentration, starting from 50% ethanol and moving up to washing with an absolute ethanol and then drying by using a critical point dryer. In the case of clusteroids within an alginate film the 5 mm × 5 mm sample was washed with deionised water and deposited on the SEM stub and freeze dried at critical point temperature. The samples were imaged using a scanning electron microscope SEM (ZEISS EVO 60 EP-SEM).