The use of solvents improved the cleaning efficacy, but filling remnants were found in three thirds of the specimens treated with xylol
and orange oil.
Tissues were dehydrated, cleaned in xylol
, embedded in paraffin blocks, sectioned at 5-[micro]m sizes, and stained with Harris' hematoxylin-eosin.
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.
At diafanization, to make the tissue semi-transparent, the alcohol present in the tissues was replaced by xylol
and the samples were kept in alcohol: xylol
(1:1) for one hour and then immersed in two sequences of xylol
for 30 minutes each.
and warm paraffin were then added into the tube.
Immunohistochemical staining was used to detect the distribution of transplanted BMSCs in the hippocampus, as follows: deparaffinization of samples in xylol
(Merck, Germany), hydration in a graded alcohol series (100, 96, 80, and 70%), incubation in 50% formamide (Merck, Germany), incubation in 2x standard sodium citrate (SSC, Merck, Germany) for 2h at 65[degrees]C, incubation in 2N HCl (Merck, Germany) for 30 min at 37[degrees]C, rinse in 0.
They were kept in a drying oven at +80[degrees]C for 20 min, passed through a pure xylol
series for 20 min and then through an ethyl alcohol series (99.
Tissue sections were dehydrated (2 x 1 min 70% ethanol, 2 x 1 min 96% ethanol, 2 x 1 min 100% ethanol, 2 x 5 min xylol
, 1 x 2 min xylol
) and embedded using Entellan (Merck).
This involved tissue slides being deparaffinised, rinsed with xylol
and then rehydrated with alcohols of different grades (absolute ethanol, 90, 80, 70, 50%).
112) Still, other studies identify xylene and chloroform (93) or just chloroform (compared with xylol
after tissue digestion with trypsin) (113) as superior clearing agents.
Various body parts were separated and mounted in Canada balsam after a brief dip in xylol
After that, they were washed in running water, dehydrated in increasing concentrations of alcohol to 100%, diaphanized in xylol
and embedded in Paraplast[R].
These mice were sacrificed and the specimens from testicular tissues were fixed in 10% neutral buffer formalin dehydrated in ascending grades of ethanol alcohols cleared in xylol
casted blocked cut at 2-5 m thickness and stained with hematoxylin-eosin for microscopic examination.
The trimmed tissues were first washed with tap water followed by dehydration through a graded alcohol series and then passed though xylol
and paraffin series before being embedded in paraffin.
Staining was made by using safranine, cleared in xylol
and mounted in Canada balsam (Ruzin, 1999).