Dehydration was performed with graded alcohol and clarification with
xylol. Finally they were covered with entellan.
(1997) performed the MATS test for
xylol, an organic non-polar solvent, and verified that the microorganisms studied were relatively hydrophilic, with solvent adhesion percentages varying between 2.7 and 26.5%.
The whole liver tissue samples were then put in an automated tissue processer Leica TP10202 for 24 hours for dehydration using alcohol and clearing using
xylol. The liver samples were then embedded in paraffin wax and cut into sections of 5 [micro]m thickness, mounted on clean glass slides coated with Mayer's egg albumin, and were stained with hematoxylin and eosin (H&E).
In the four experimental groups, canals were retreated with PTR alone or in combination with
xylol, orange oil, and eucalyptol.
Before staining, samples were dewaxed with
xylol and dehydrated with alcohol.
These were placed in the oven at 58[degrees]C, for 10 min, followed by deparaffinization in
xylol and rehydration in alcohol at decreasing concentrations, and washed in distilled water and PBS (0.1 M sodium phosphate buffer, pH 7.2), for 10 min.
The latter was eliminated by
xylol before being flowed in molds containing paraffin melted by heating at 60[degrees]C [23].
Light microscopy: The fixed ovaries from cultured (n=5 in each group) and non-cultured groups (n=5 for 14 day mice and n=5 for 21 day mice) by Bouin's solution were dehydrated in a graded series of ethanol and cleared with
xylol and finally embedded in paraffin wax.
After fixation, the samples were dehydrated in ascending concentrations (70, 80, 90 and absolute) of ethyl alcohol, followed by diaphanization with
xylol and inclusion in paraffin.
Liver tissue purification process has been carried out in the next stage of
xylol and paraffin embedding.
Tissues were dehydrated, cleaned in
xylol, embedded in paraffin blocks, sectioned at 5-[micro]m sizes, and stained with Harris' hematoxylin-eosin.
3-5 slices of 5 micron from selected samples placed in the 1.5 ml micro- tube and DNA samples were extracted with phenol-chloroform manually with using
xylol, ethanol, Layzyz buffer , proteinase K, phenol saturated, chloroform isoamyl alcohol, sodium acetate, isopropanol and the finally 50 micro- liter of distilled water added.