Imaging human keratinocytes grown on electrospun mats by scanning electron microscopy

Drying is a critical step in preparing cell samples for examination with scanning electron microscopy (SEM). The two commonly used drying procedures are the critical point drying (CPD) and the chemical drying using hexamethyldisilazane (HMDS drying). Here we compared the application of these procedures for the drying of HaCaT human keratinocyte cells grown on electrospun nylon mats. Both drying procedures allowed us to obtain images of the cells and characterize the microvilli on the cell surface. After HMDS drying the membrane was less damaged than after CPD. Both drying procedures could be used to investigate contact guidance-the substrate-induced changes in cell shape. The aspect ratio of HaCaT cells grown on the aligned and random mats was 4.2 ± 2.8 and 1.5 ± 0.3, respectively.