Comparison of Environmental Scanning Electron Microscopy in Low Vacuum or wet mode for the investigation of cell biomaterial interactions

摘要: Aim : The aim of the present study was to investigate efficacy environmental scanning electron microscopy (ESEM), in low vacuum mode (LV-ESEM) and wet (wet-ESEM) assessment cell-material interactions. Methods Mouse calvaria MC3T3 cells (ATCC) were seeded on commercially pure machined titanium discs 10 mm diameter Dulbecco modified MEM, 10% Fetal Bovine Serum, 1% Penicillin Streptomycin Glutamine. Samples then processed for microscope observation by rinse Phosphate Buffer saline fixation 4.5% Glutaraldehyde. rinsed Sodium Cacodylate buffer observed or dehydrated alcohol prior LV-ESEM observation. Fresh samples 0.9% NaCl solution wet- ESEM. Results No significant loss detail when non analysed at LV-ESEM.The fresh wet-ESEM however proved difficult need eliminate water which forms a layer covering sample, thus hiding cell surface details. When reducing vapor pressure chamber, evaporated immediately started precipitate collapsed, no further investigation possible. Conclusions use vacuum-ESEM after fixation, but without dehydration gold sputter coating viable alternative traditional high SEM