The samples were deparaffinized in a graded series of xylol
and rehydrated in a graded series of alcohol.
The tissue slices were processed in various grades of alcohol and xylol
and subsequently embedded in paraffin wax.
Tissue samples were passed from alcohol series for dehydration and from xylol
series for clarification then blocked in paraffin.
-removal of paraffin by Xylol
, increasing hydration by ethanol alcohol, inhibition of peroxidases in 3% methanol solution, two times of washing with PBS-T solution, Antigen retrieval using Dako solution, two times of washing with PBS-T solution, Blocking for one hour at 37 degrees using secondary antibody host serum (ab97245), Goat Anti Mouse (HRP), two times of washing with PBS-T solution.
Sections were passed through alcohol and xylol
series, and then adhesive (Entellan[R]) was dropped, and lamella was applied.
After sectioning, the sections were de-paraffinized in xylol
followed by hydration in descending grades of alcohol (100%, 95%, 80%, and 70%) and distilled water.
Finally, the slides were counterstained with hematoxylin, washed in running water, dehydrated in a sequential series of alcohol and xylol
baths, and covered with coverslips mounted with Permount[R] (Fisher Scientific, USA).
Routine pathology tissue procedures were applied with the tissues passed through a graded series of alcohol (50%, 75%, 96%, 100%) and xylol
, then embedded in paraffin blocks.
Then, they were dehydrated in a series of alcohols, clarified in xylol
and embedded in Paraplast (Paraplast Plus embedding medium; melting point: 54 [degrees]C; Sigma-Aldrich Chemical Co., St Louis, MO, USA).
After placing into xylol
, the cross-sections were embedded in paraffin.
The samples were then dehydrated, cleared with xylol
, embedded in paraffin, sectioned (4 [micro]m thickness), and stained with hematoxylin and Ehrlich's eosin.
Abdomens then were washed in water, dehydrated in a series of increasingly concentrated ethyl alcohol solutions, and cleared in xylol
. Abdomens were dissected to remove the tergites with tergal glands and to separate the genitalia.