Post fixation was made in 1 % osmium tetroxide
buffered to PH 7.
Geuna, "A simple protocol for paraffin-embedded myelin sheath staining with osmium tetroxide
for light microscope observation," Microscopy Research and Technique, vol.
For TEM analysis, the samples were processed as described for SEM procedures up to the fixation in osmium tetroxide
1% for two hours.
For scanning electron microscopy, formalin-fixed lung tissues were trimmed into 2-4-mm pieces, postfixed for 1 h in 1% osmium tetroxide
, and rinsed for 30 min in 0.
The polybutadiene phase was stained with osmium tetroxide
2 (three times for 10 minutes), post-fixed in 1% aqueous osmium tetroxide
solution for 1 hour, washed twice for 15 minutes in distilled water, transferred to a 0.
1 M phosphate buffer, post-fixed in 1% osmium tetroxide
, dehydrated in alcohol, embedded in PolyBed 812 (Polysciences Inc.
For scanning electron microscopy (SEM) the material was fixed with glutaraldehyde on phosphate buffer followed by complete fixation in 1% solution of osmium tetroxide
in the same buffer, subjected to dehydration in alcohol, acetone, then dried by critical point in the apparatus of the NDS-2 and sputtered with gold in the apparatus IB-2.
4, 2 h, 4[degrees]C) and post-fixed in presence of osmium tetroxide
(2% osmium tetroxide
in Milloning buffer 0.
2, for 3h, post-fixed in 1% osmium tetroxide
in phosphate buffer, washed in the same buffer, dehydrated in a growing acetone series and embedded in Araldite resin (Durcupan ACM, Fluka, Sigma-Aldrich, St.
The later range from formalaldehyde, gluteraldehyde, to osmium tetroxide
solutions, depending on type of tissue and purpose.
The technique uses a 1% solution of osmium tetroxide