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).
