Cells were washed in PBS and counterstained
with Hoechst 33342 nucleic acid stain for 10 minutes.
with diluted hematoxylin or hemalaun (blue).
Some of the tissue sections from the heart, lungs, liver, and spleen were stained with alum hematoxylin (Harris hematoxylin) and counterstained
with 90% ethanolic extract of Z.
A total of 10-15 metaphase spreads were analyzed, using a fluorescence microscope (AxioImager.Z1 mot, Zeiss) equipped with appropriate filter sets to discriminate between a maximum of five fluorochromes and the counterstain
Briefly, conjunctival scrape smears were covered with 30 microliters of Evans blue counterstain
containing solution of fluorescein-isothiocyanate(FITC-) conjugated antibodies for 20min at 20[degrees]C in a dark, humidified chamber.
Slides were washed with PBS and mounted in Vectashield Mounting Medium (Vector Labs) containing the dye 4,6-diamido-2-phenylindole (DAPI) to counterstain
The analysis of antibody binding was performed with a diaminobenzidine (DAB) kit, and hematoxylin was used as the counterstain
. The slices were dehydrated through graded ethanol and xylene and mounted with Entellan.
Hematoxylin was applied for 2 minutes and washed with distilled water.
DAB was used as the chromogen, and Mayer hematoxylin was used as the counterstain
. The positive control was normal testis tissue, which shows cytoplasmic staining in Leydig cells.
Acid fuchsin counterstain
(5 mL of 1% acid fuchsin + 95 mL of picric acid + 0.25 mL of 12 N hydrochloric acid) was added to the cell layers for 5 minutes.
The slides were developed in 3,37-diaminobenzidine (DAB), followed by a haematoxylin counterstain
. Sections from nonneoplastic colonic mucosa from non-FAP patients, normal breast, and endometrium were used as positive controls, and for each case a section from which the primary antibody was replaced by phosphate buffered saline was used as a negative control.
To remove residual counterstain
, cells were washed twice in PBS before mounting.