Hematoxylin and Eosin

Hematoxylin:
Hematoxylin is extracted from the heartwood
(‘logwood’) of the tree (Hematoxylon-
Campechianum ), that originated in the Mexican
State of Campeche, but which is now mainly
cultivated in the West India.
Hematoxylin itself is not a stain, oxidization
process must be done to obtained the color
(hematin).
Objective:
To differentiate the tumor / cancer or benign cells
from autopsy and biopsy.
To identify cell constituents demonstrated with H &
E stain.

Principle:
H stains the acidic parts of the cells (nucleus), so its
called as nuclear stain, while eosin acts as a acidic
stain and bind with the basic part (cytoplasm).
Classification according to method of
oxidation( Ripening):
1. Natural oxidation : carried out by exposure to
light and air. Very slow take about 3-4 months.
Lasts for longer.

2. Chemical oxidation: use oxidizing agents to

converts hematoxylin to hematin.
Take short time to prepare, ready for use after
preparation.
According to which mordant is used:
a) Alum Hematoxylin
b) Iron Hematoxylin
c) Tungsten Hematoxylin
d) Molybdenum Hematoxylin
e) Lead Hematoxylin
f) Hematoxylin without mordant.
Alum hematoxylin:
Used routinely in the hematoxylin and eosin stain and
produce good nuclear staining.
The mordant is aluminum:
Alum- aluminum potassium sulfate
Alum- aluminum amonium sulfate

A.H can be used progressively or regressively.

Calssification of Alum hematxylin:
Ehrilch`s Hematoxylin .
Mayer`s H.
Harris H.
Ehrilch`s Hematoxylin :
Naturally ripened strong alum Hematoxylin.
Stain mucin, salivary glands and cartilage. suitable
for tissue subjected to acid decalcification.
Suitable for tissue that had been stored for a long
period in formalin ( tissue become acidic over the
storage period).
Not ideal for frozen section.
Ripened by sodium iodide, in case of emergency.
Mayer`s H :
Ripened with sodium iodide.
Used for regressive and progressive stain.
Used as counter stain in the demonstration of
glycogen (PAS).
Harris`s H:
Ripened with mercuric oxide.
Used as regressive stain in routine histology lab.
Used as progressive stain in diagnostic exfoliative
cytology .
EOSIN
Eosin : is xanthene dyes.
*Examples of easily obtainable
commercially
Eosin Y (yellowish, water &alcoholic
soluble):most widely used.
Eosin B(bluish, erythrosine B), & Ethyl
eosin(alcohol soluble):are rarely used.
ALTERNATIVE RED DYES
FOR EOSIN :
PHLOXINE
BIEBRICH SCARLET