The usual sequence among the commoner hypogene (deposited by generally ascending solutions) metallic sulfide minerals is pyrite, enargite, tetrahedrite, sphalerite, chalcopyrite, bornite, galena, and pyrargyrite.
Briefly, specimens were retrieved eight weeks after implantation, fixed in 10% formalin solution, dehydrated in an ascending series of ethanol solutions, and embedded in paraffin.
For cryosectioned tissue, brains were first cryoprotected through an ascending series of sucrose solutions until they sank (10, 20, 30% sucrose w/v in PBS).
Vibratome sections containing this region were post-fixed in 1% osmium tetroxide for 45 minutes and dehydrated through an ascending series of ethanol solutions (including dehydration for 40 mins in 70% alcohol containing 1% uranyl acetate) and propylene oxide.
The samples were then dehydrated through an ascending series of ethanol solutions (2×30 min each in 70% and 96%, 2×20 min in 100%), followed by 2×1 h in chloroform.
Following dehydration through an ascending series of ethanol solutions and propylene oxide, the nerves were embedded in Durcupan resin overnight and polymerised for 48 hours at 50°C.
After hybridization, the sections were washed in 2 × SSC at 50°C and then dehydrated in an ascending series of ethanol solutions containing 0.3 mol/l ammonium acetate.
The PFA was rinsed out with several changes of phosphate buffered saline (PBS), and embryos were taken through an ascending series of sucrose solutions (7, 15, 30%).
They were rinsed in DIW and dehydrated in an ascending series of ethanol solutions to 100% ethanol, rinsed twice in acetonitrile, and embedded in quetol epoxy resin.
For electron microscopy, after development of the immunoperoxidase reaction, sections were first treated with 1% OsO4 in 0.1 m PB for 10 min in the dark, on ice, and then dehydrated in an ascending series of ethanol solutions, followed by acetonitrile.
Our findings suggest that the elasticity change of tissues in the formalin solution has an ascending trend, but that of Thiel remains almost constant, providing a more real texture and properties.
