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.
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.