For morphological analysis of the cell attached scaffolds, the samples were washed with PBS after removing the medium and fixed with 0.6 ml of 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer solution for 30 min at 4 °C. After thorough washing twice with PBS, scaffolds were dehydrated through a series of graded ethanol concentrations (30%, 50%, 70%, 90% and 100%) and finally dried in HMDS. After proper drying, scaffolds were coated with gold and palladium using a sputter coater (Cressington 108 Auto) and were then observed using an SEM (FEI Quanta 3D FEG dual beam ESEM, USA) operated at an accelerating voltage of 5 kV.
108 auto
Manufactured by Thermo Fisher Scientific
Sourced in United States
The 108 Auto is a laboratory instrument designed for automated sample processing. It is capable of performing basic laboratory functions such as mixing, diluting, and aliquoting samples. The 108 Auto is a compact and versatile device suitable for use in various laboratory settings.
Automatically generated - may contain errors
3 protocols using 108 auto
1
Morphological Analysis of Cell-Attached Scaffolds
2
Morphological Analysis of Cell-Attached Scaffolds
3
SEM Imaging of Fixed Cells
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For SEM experiments, cells were cultured and manipulated as described above to illustrate different conditions/surfaces. Cells were then fixed with 2% PFA, 2.5% glutaraldehyde in PBS for 1 h at room temperature. Fixed samples were dehydrated by exposure to a graded series of water-ethanol mixtures. Once in 100% ethanol, samples were dried at the critical point of CO₂ using a critical point drying technique (Tousimis Samdri-795) to preserve structural details. Dried samples were then coated with 10 nm of gold palladium alloy using a sputter coater (Cressington 108 Auto), and imaged using a FEI Helios 600 NanolabScanning Electron Microscope working at an accelerating voltage of 5 kV.
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