After washing with PBS, cells were counterstained
with Hoechst 33342 nucleic acid stain (Thermo Fisher Scientific-Molecular Probes) for 10 minutes.
The IHC sections were counterstained
using Harris hematoxylin.
To determine the number of proliferating microglia in the OB we performed triple fluorescence staining labelling for Iba1 and MCM2, including DAPI as nuclear counterstain
. Therefore, we adapted the protocol for MCM2 from above and extended the blocking step to 3 h in 3% donkey serum (in TBS-T).
When Z.officinale extract was used as a counterstain
for hematoxylin, the nuclei took deep green coloration, which enabled a clear contrast to be made between the different structures of the cells.
A drop of lactophenol blue was added as the counterstain
. The images of filamentous fungi were observed under light microscope.
Captured cells are then further qualified as CTCs on the basis of expression of cytokeratins (CK), nuclear counterstain
with 4',6-diamidino-2phenylindole (DAPI), and absence of CD45 expression (11, 12).
Diaminobenzidine was used as the final chromogen, and hematoxylin was used as a nuclear counterstain
For the most part, the vast majority of microscopic pathology is still done by viewing or imaging one marker at a time in serial sections, using a hematoxylin counterstain
and either eosin (H&E) or, for protein IF1C measurements, 3,3'-diaminobenzidine (DAB) as a chromogen.
Mayer's hematoxylin was used as the counterstain
. Prostatic glandular basal cells and duodenal mucosal tissue were used as the positive controls for maspin and ezrin, respectively.
Appropriate Alexa Fluor 488 or 594 F (ab7)2 fragment of goat [alpha]-rabbit or [alpha]-mouse IgG (H+L) antibodies or Alexa Fluor 555 goat [alpha]-rabbit IgG (H+L) or Alexa Fluor 594 F (ab7)2 fragment of rabbit [alpha]-goat IgG (H+L) (A-11070, A-11020 or A-21428, A-21223; 1: 2000; Invitrogen) and DAPI (1 [micro]g/mL) counterstain
were used for visualization.
Streptavidin-biotin method was utilized to perform staining, with DABB being used as the chromogen and haematoxylin as counterstain
The dark brown dye 3,3,-diaminobenzidine (DAB) is commonly used as an antibody-specific stain whereas the blue dye, hematoxylin, is used as a counterstain
and is unspecific in its binding.