Scanning Electron Microscopy
Scaffolds were fixed in 100 µl fixing solution (2.5 M glutaraldehyde,
0.2 M phosphate buffer, 1 mM sucrose solution) for 2 hours at room
temperature, washed three times with washing buffer (0.05 M sucrose,
0.05 M phosphate buffer, pH 7.2) for 10 minutes. Samples were dried by
liquid displacement using an ethanol: water gradient (25%, 50%, 70%,
80%, 95%, 100%) and then ethanol: hexamethyldisilazane (3:1, 1:1,
1:3). Finally, samples were air dried overnight. Dried samples were
sputter coated with gold/palladium, mounted and imaged on a LEO 1550
scanning electron microscope.