Images were performed by fluorescence microscope (Nikon Corporation, Japan) after the cells were incubated in 3,3'-diaminobenzidine (Beijing OriGene Technology Co., Ltd.,) for 5 min and stained in hemalum
, and all images were measured by Image Pro-Plus 6.0 software (Media Cybernetics, Silver Spring, MD, USA).
Cell nuclei were counterstained by hemalum
within 2 min.
Puchtler, ""Harris hematoxylin," what harris really wrote and the mechanism of hemalum
stains," Journal of Histotechnology, vol.
Counterstaining was performed for 30 sec with Mayer's hemalum
solution (Merck Darmstadt, Germany).
The specimens were counterstained with hemalum
. The stained specimens were viewed at an objective magnification of x200 and x400 by three investigators.
After paraffin embedding, 5[micro]m sections were cut and stained with hemalum
phloxine saffron (HPS) for histological evaluation.
Sections were counterstained with Mayer's hemalum
and rinsed in distilled water.
The sections were lightly counterstained with hemalum
and luxol blue.
Binding was detected by using monoclonal antibody 2G3 against RHDV, biotinylated anti-mouse IgG, horseradish peroxidase-conjugated avidin, and 3-amino-9-ethylcarbazole substrate with hemalum
counterstaining, as described (2).
Posteriormente se realizo una contracoloracion con hemalum
de Mayer, se lavaron con agua destilada, dejando secar los cortes y se realizo el montaje en Entellan.
Sections were then counterstained with hemalum
(progressive Mayer's) solution (Sass, 1958) for two min.
After counterstaining with hemalum
and dehydrating were carried out, the slides were placed under coverslips with Neomount (VWR, Vienna, Austria).