Scaffold Characterization by SEM and CLSM

The microscopic view of the scaffolds was performed using scanning electron microscopy (SEM) for qualitative analysis of the osteoblast cells along with determination of the pore dimensions of the scaffolds. For assessments using CLSM, colonized cells present on the scaffolds were fixed using 3.7% paraformaldehyde for 20 min. Cell cytoskeletal filamentous actin (F-actin) was visualized by Alexa Fluor 488 Phalloidin (1:25 dilution in PBS, 1.5 h) treatment of cells and counter-staining with propidium iodide (1 μg mL⁻¹, 20 min) for labelling of cell nuclei. The cultures were then placed in Vectashield and assessed using a Leica SP2 AOBS (Leica Microsystems, Wetzlar, Germany) microscope [28 (link)].