Imaging human keratinocytes grown on electrospun mats by scanning electron microscopy

Abstract

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.

Nanogallium-poly(L-lactide) Composites with Contact Antibacterial Action

“…After two hours of soaking in the fixative at room temperature, HaCaT cells were transferred together with the films into 0.6 mL of DPBS, which was exchanged with 0.5 mL of fresh DPBS after 5 min and once again after 5 min. Then, gradual dehydration followed, with the sequential introduction and removal of the following liquids (in 0.5 mL amounts) [ 68 ]: 10%, 20%, 30%, 40%, 50%, 60%, 70%, 70%, 80%, 100%, 100% ethanol, 1:1 (1,1,1,3,3,3-Hexamethyldisilazane (HMDS)):ethanol mixture and 100% HMDS. The samples were air-dried from HMDS, glued onto carbon tape, and coated with 6 nm of Pt before observation under SEM.…”

Alginate/pectin dressing with niosomal mangosteen extract for enhanced wound healing: evaluating skin irritation by structure-activity relationship

“…Sterilized glass coverslips were used as controls. After a 24-h r incubation, all samples were fixed with 3% glutaraldehyde (Sigma–Aldrich, MO, USA) for 15 min, dehydrated with an ethanol series [ 49 , 50 ], and treated with hexamethyldisilazane (Sigma–Aldrich, MO, USA) for 5 min [ 51 ]. The samples were sputter-coated with gold and examined using a scanning electron microscope (SEM; JCM-7000; JEOL, Tokyo, Japan).…”

The Hirudo Medicinalis Microbiome Is a Source of New Antimicrobial Peptides

“…Then, samples were dehydrated with increasing ethanol percentages (10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 96% for 15 min in each solution). Then, the samples were incubated in a hexamethyldisilazane (HMDS, Reachem, Russia): ethanol mixture (1:1 by volume) for 10 min and in 100% HMDS overnight until complete HMDS evaporation [ 55 ]. After chemical drying, the samples were covered by a 10 nm gold-palladium alloy using Sputter Coater Q150T (Quorum Technologies, Lewes, UK).…”