Sigma field emission sem fesem

In culture, 4MU was dissolved in DMSO and was added at 100 µg/ml (approximately 300 µM). For in vivo studies, 4MU (Alfa Aesar, Haverhill, MA) was pressed into the mouse chow by TestDiet® as previously described (63) . Mice were initiated on the 4MU chow at 6 weeks of age and were maintained on this diet until euthanized, unless otherwise noted.
Scanning Electron Microscopy (SEM): BMDC were isolated as described above, and grown on fibronectin-coated glass slides for 24 hours, were fixed overnight in 4% paraformaldehyde with 2% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.4). The slides were gently washed twice with the same buffer, and then post-fixed in 1% aqueous osmium tetroxide (OsO4) for one hour. Samples were then washed twice in purified water and dehydrated in an increasing ethanol series (50%, 70%, 80%, 90%, 100%, 100%, 5 min each). Finally, the specimens were critical-point dried (CPD) in liquid CO2, in a Tousimis 815B critical-point dryer (Tousimis, MD). CPD-dried samples were mounted on SEM stubs with adhesive copper tape and sputter-coated with 4 nm of Au/Pd film (4 nm in thickness) in a Denton Desk II machine (Denton Vacuum, NJ). The prepared SEM samples were examined with a Zeiss Sigma field emission SEM (FESEM) (Carl Zeiss Microscopy, NY) operated at 2-3 kV, using InLens SE detection.