Scanning
electron microscopy
visualize
BOEC
polarity
differentiation level
polyester
membranes
hanging
inserts
monolayers
washed twice
with Krebs
solution
fixed
glutaraldehyde
rinsed
buffer
containing
saccharose
insert membranes
removed
polystyrene
housing
placed
plate
containing
cacodylate
buffer.
Subsequently, cells were dehydrated in an
ascending series of ethanol concentrations (70%, 90%, 95%
each for 15 minutes at room temperature, and 100%, three
times, for 30 minutes) and submitted to critical point drying
in a Leica EM CPD030 after which the monolayers were
mounted on a stub and gold coated in a sputter coater.
Scanning electron microscopy imagingwas performed with
an SEM 515 (Philips, The Netherlands) at different magnifications.
Monolayers were assessed on cell polarity, the
presence of microvilli and cilia, monolayer integrity, and cell
growth [35]. For each treatment, two monolayers were
processed, and all results are descriptive, not quantified.