Aqueous Hematoxylin

Aqueous Hematoxylin

Biømeda’s hematoxylin is water- based, unlike the most common hematoxylins available which are alcohol-based. The use of alcohol-based hematoxylin destroys most of the chromogens (i.e., AEC, Fast Red, BCIP/INT, etc.) used for immunocyto-chemistry. Biømeda’s hematoxylin formulation avoids this problem and is specifically designed for nuclear counterstaining using automated and/or manual immunocytochemistry techniques. This hematoxylin provides a clean, strong and permanent nuclear stain in the formalin-fixed, paraffin-embedded tissue sections, as well as in frozen tissue sections.

Aqueous hematoxylin was designed to overcome one of the major obstacles in immunocytochemistry: the use of alcohol-based hematoxylins which destroy many of the most useful chromogens used for both immunoperoxidase and immunoalkaline phosphatase assays. Although alcohol-based hematoxylins, such as the Harris formula, produce rapid and selective nuclear stains, their application results in the rapid decolorization of the brown-red oxidation product of 3-amino-9-ethylcarbazole (AEC). This is unfortunate because AEC is not only the most powerful chromogen available for immunoperoxidase techniques but its brown-red product contrasts far better with blue nuclear stains than does alcohol-stable chromogens such as diaminobenzidine (DAB). This problem exists also with binary alkaline phosphatase chromogens such as 5-bromo-4-chloro-3-indolyl phosphate and iodonitrotetrazolium (BCIP/INT). Biømeda’s hematoxylin provides a clean, strong, superior nuclear stain in only one minute which is 8-10 times faster than other water-based hematoxylins such as Mayer’s formulation. When counterstaining specimens subjected to in situ DNA hybridization procedures, we recommend using Biømeda’s new Probe/Hematoxylin which provides controlled nuclear counterstaining for better contrast and stain definition.

Store at room temperature.

Technical Information:
Can dilute in water, but check to be sure to keep pH in balance. If too concentrated, dip for 5-10 seconds instead of recommended 2 minute incubation. Once completed staining, there is NO problem using alcohol or other organic solutions to wash, etc. pH should be approx. 2-3.

Seven Step Procedure: 1.    After the immunostaining, rinse slides with distilled water and drain.
2.    Introduce the hematoxylin.
3.    Incubate the slides for 1-2 min. at room temperature.
4.    Drain off the hematoxylin and wash slides several times with distilled water.
5.    Incubate the slides with 1X Automation Buffer or PBS (pH7.4) for 1 minute.
6.    Wash with distilled water.
7.    Mount with Crystal/Mount (Cat. #M02 or #M03).
Do not use beyond the expiration date stated on the label.

For Research Use Only.

This is a laboratory reagent and should only be used by trained laboratory personnel. Avoid contact with skin or clothing.