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6.
7.
Instructions for use
Hemalum solution acidic according to Mayer
T865
The dye precursor for hemalum solution acidic acc. to Mayer is hematoxylin, a natural dye which initially has to be
converted by oxidation to hematein (ripening). The hemalum dye itself subsequently emerges by steeping it with
alum.
Hemalum stains cell nuclei blue. The bonding of the dye must be carried out in a highly acidic setting.
Subsequently a pH-value of more than 7 is needed to maintain the blue colour. This can be achieved by rinsing
intensively with tap water (blueing). In this way you also increase the permanency of the staining as the dye is
hardly soluble in alkaline medium.
Hemalum solution acidic according to Mayer is used, for example, for nuclei staining in H&E staining (see below).
Hematoxylin-eosin staining (H&E staining)
H&E staining is a standard staining method in histology. It provides an overview of the different tissue structures
often allowing an immediate diagnosis.
After hemalum staining, Eosin Y solution 0.5 % in water is used for counterstaining.
Adding 1 drop of pure acetic acid per 100 ml terminates alkaline blueing and makes contrast staining easier.
Additional chemicals required:
Eosin Y solution 0.5 % in water (Art. No. X883)
Pure acetic acid, p.a. (Art. No. 3738)
Ethanol denatured: 99.8 % (Art. No. K928), 96 % (T171), 70 % (T913)
Clearing Agents: ROTI®Histol (Art. No. 6640)
or ROTICLEAR® (Art. No. A538)
or xylene p.a. (Art. No. 4436)
Mounting Media: ROTI®Histokitt (Art. No. 6638), compatible with ROTI®Histol
or ROTI®Mount (Art. No. HP68), compatible with ROTICLEAR®
or ROTI®Histokitt II (Art. No. T160), xylene compatible
Instructions*:
Use de-waxed or frozen sections on slides wetted in distilled water:
1. Stain with Hemalum solution acc. to Mayer
5-10 min
2. Rinse quickly with distilled water
3. Blue in flowing tap water 15 min
4. Rinse with distilled water 2 min 8. Clear with clearing agent
9. Mount with appropriate mounting medium
* Acc. to Romeis, Mikroskopische Technik, 18. Auflage, Spektrum Akademischer Verlag (2010)
Results:
Cell nuclei: blue
All other structures: reddish with different intensity (e.g. cytoplasm, collagen fibres, erythrocytes)
Note:
The colour intensity depends on the pre-treatment and the composition of the samples to be stained.
It may initially be necessary to adapt the method to the respective conditions.
5. Incubate in Eosin Y solution
(add 1 drop of pure acetic acid per 100 ml solution) 1-5 min
Rinse quickly with distilled water
Dehydrate by ascending alcohol series (70-96-100 %)
g Warning H302-H315-H319
Carl Roth GmbH + Co. KG
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P.O. Box 100121 • 76231 Karlsruhe
Phone: +49 (0) 721/ 5606-0
Fax: +49 (0) 721/ 5606-149
[email protected] • www.carlroth.com sse 06/2021
The com pany is a l imited partnersh ip with headquarter s in Karlsruhe, reg. court
Mannheim HRA 100 055. Roth Chemie GmbH, with headquarters in Karlsruhe, reg.
court M annheim HRB 100428, is the personally liable partner. Managi ng Directo r:
André H oudelet. Sales tax i dentification n umber: DE 143621073.
Hemalum solution acidic acc. to Mayer
T865.1 500 ml
T865.2 1 l
T865.3 2,5 l
SDB-Version:05.10.2021
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