Has someone of you experiences with different fixation methods of HUVECs as described above? We have an immunofluorescence protocol for heparan sulfate staining (including fixation with 0,5% glutaraldehyde in PBS), but the structure doesn't look like as we would expect. Perhaps it's a problem of the fixation of heparan sulfate...
Damien G Harkin
I agree. Use 2-4% (w/v) formaldehyde (pH 7.2) for 20 minutes.
Glutaraldehyde is a better fixative (benefits not really apparent unless using electron microscopy), but produces more autofluorescence.
Can also compare with alcohol based techniques if problems persist.
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