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Hematoxylin counterstain

Manufactured by Roche

Hematoxylin Counterstain is a laboratory reagent used in histology and cytology. It is a dye that stains nuclei in cells, providing contrast to the primary stain. The core function of Hematoxylin Counterstain is to enhance the visual distinction between cellular structures during microscopic examination.

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2 protocols using hematoxylin counterstain

1

Immunohistochemical Staining of CYP27A1 in Liver

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The liver and heart tissues were stained with H&E. Immunohistochemical (IHC) staining for the CYP27A1 protein in liver tissue was conducted by using an autostain system with exclusive reagents (Discovery XT system; Ventana–Roche Diagnostics K.K., Basel, Switzerland). Deparaffinized 5-µm thick specimens of hepatic tissue were incubated with the primary antibody for anti-CYP27A1 (1:100) for 1 hour at room temperature, and then incubated with a biotin-conjugated universal secondary antibody (Ventana) for 32 min at 37 °C. It was developed using the DAB map kit (Ventana). The nucleus and cytoplasm were stained by using Hematoxylin Counterstain (Ventana) and a bluing reagent (Ventana). Specific immunoreaction of the primary antibody was confirmed by incubation without the primary antibody.
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2

Histological Analysis of SELP in Liver and Lungs

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Liver and lungs of each animal were immediately fixed in 10% buffered formalin for 48 hrs and then transferred to 70% ethanol. Sections from each liver lobe and each lung lobe were cut in 5 μm slices and analyzed. Tissue samples were stained using both hematoxylin and eosin (H&E) stain and an immunohistochemistry (IHC) stain specific for SELP using an antibody for poly-Histidine (anti-6X His tag®, Abcam, Cambridge, MA; 1:3000 dilution), which is found in the carboxyl-terminal sequence of the SELP polymer strand. A chromogenic detection was used, IView diaminobenzidine research detection kit (Ventana Medical Systems) and hematoxylin counterstain (Ventana Medical Systems) in the IHC procedure. A piece of pre-gelled SELP was fixed, sliced, mounted onto a slide, and stained as a positive control using both stains. Slides were imaged using a Nikon DXM 1200C Digital Camera affixed to an Olympus BH2 microscope and analyzed in ACT-1C for DXM1200C software.
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