7. Glutaraldehyde is a slow fixative and preserves cell morphology
well. It may be stored at À20
C and can be used for up to a
week before it will start to crosslink.
8. Don’t let the samples dehydrate during SEM sample preparation, as this may affect the cell morphology. The steps are best
performed in a well plate. It is not necessary to remove the
liquid entirely, as multiple washes are performed.
9. If necessary the dehydration process may be paused at 70% and
the samples left at 70% overnight; otherwise the process needs
to be backtracked.
10. Depending on the materials used for the scaffolds, the samples
may be dried in a critical point dryer following the 100%
ethanol incubation.
11. The samples for CLSM may be fixed and stored in PBS at 4
C
and stained at a later time.
Fig. 6 Primary human dermal fibroblasts on a MEW scaffold with fiber spacing of
0.2 mm. The samples were prepared according to the above protocol and
imaged using a CLSM. The cell nuclei were stained with DAPI and the actin
filaments with FITC-conjugated phalloidin
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