The cell suspension was stained with 5 [micro]L of lacto-acetic orcein
, covered with a cover slide and sealed with glue.
Tissues were processed for routine histology and 5-7 micro meter thick sections were cut and were stained with hematoxylin and eosin, Orcein
and Masson's trichrome stains.
Meristematic regions with 1 mm of length excised and squashed in a drop of 1% acetic orcein
mixed with a drop of 45% acetic water (Jahier 1992).
The samples were then stained with 1% (w/v) orcein
in 45% (v/v) acetic acid solution and examined under a phase-contrast microscope (IX-50, Olympus, Tokyo, Japan) at 400x.
Oocytes were stained with 1% aceto-orcein
in 45% glacial acetic acid).
Permanent slides were made after softening the tissues in 45% acetic acid, drying on a hot plate and staining with lacto-acetic orcein
for 3 to 5 min.
Application of pectinase and cellulase in an orcein
The root-tips were stained in 2% lactopropionic orcein
after cold hydrolysis in 5N HCl for 10 min and were squashed in 45% lactopropionic acid.
Metaphase I configurations were analyzed by squashing a small sample of testicular tissue stained with acetic orcein
For this study, 3 new histologic sections (6 [micro]m thick) were obtained from each slide and stained with hematoxylin-eosin, Masson trichrome, or orcein
(for demonstration of elastic fibers).
A drop of fixed sample was loaded on a slide, mixed with 2-3 drops of orcein
For histological study small segments of the CS from its proximal, middle, distal parts, including the Thebesian valves were processed by routine histological techniques and 5-7 microns thick sections were cut and stained by--H and E, Masson's Trichome and Orcein
These were put in ten drops of 2% acetic orcein
stain (GUERRA; SOUZA, 2002) + one drop of 1N HCl, buckling was performed for a short time (1 to 2 minutes), just to allow hydrolysis and staining of the cell.
for morphological analysis according to Paulete and Becak (1976), Alcian Blue for detection of acid polysaccharides according to Junqueira and Junqueira (1983), Periodic Acid Schiff stain (PAS) for detection of neutral polysaccharides according to Junqueira and Junqueira (1983), Picrosirius Red for detection of collagen fibers I and III accoding to Pearse (1985) by polarized light, Orcein
for the detection of elastin fibers according to Paulete and Becak (1976).
Some of the COCs were removed and fixed with acetic alcohol (alcohol:acetic acid = 3:1, v/v) for at least 2 days at room temperature and stained with 1% acetic orcein