Confocal Microscopy Analysis of Cell Morphology

Similar to our previous report^{35 (link)}, to examine the cell morphology under confocal microscopy after 24 h of incubation, cells on each surface were fixed in 4% paraformaldehyde for 20 min. A 1 μL stock solution of phalloidin-488 (AAT Bioquest) was diluted in 100 μL of PBS containing 1% bovine serum as the main working solution for staining. The surfaces were also washed with PBS for three times before staining the nuclei with 1 μL of Hoechst 33342 (AAT Bioquest) in 1 mL of PBS buffer for 10 min. Upon completion of staining and washing, a drop of Fluoromount (NOVUS) was carefully aliquoted onto the surface and subsequently covered with a coverslip and sealed with nail varnish. The stained cells with actin filament were observed under confocal microscopy (LEICA SP8) at 40x magnification to examine the cell morphology.