Hematoxylin and Eosin Staining Method Flashcards
For routine microanatomical examination,_______is the staining method of choice
Hematoxylin and Eosin (H&E)
H&E
The two stains were independently introduced in____ and ____, respectively, by____ and ____
1865 and 1875
Böhmer and Fischer
In 1876,______ described their use in combination as a tissue staining method for staining different materials with different colours.
Wissowzky
is the most important type of bonding that occurs in histologic staining techniques.
Ionic bonding
It involves electrostatic attraction between ions of opposite charge, one of which is fixed in the tissue, and the second of which is in the dye
Ionic bonding
alone is not technically a dye, and will not directly stain tissues.
Hematoxylin
Hematoxylin needs to be used in combination with a “_____” – a compound that helps it link to the tissue
mordant
The mordant used is typically a metal____, such as aluminium.
cation
Hematoxylin in complex with aluminium salts is cationic and acts as a______ dye.
basic
Hematoxylin is______ charged and can react with______ charged, basophilic cell components, such as nucleic acids in the nucleus.
These stain____ as a result.
positively
negatively
blue
is anionic and acts as an acidic dye.
Eosin
Eosin
It is_____ charged and can react with____ charged,_____ components in the tissue, such as amino groups in proteins in the cytoplasm.
These stain____ as a result.
negatively
positively
acidophilic
pink
is a useful all-purpose stain that is also quick and easy to use. Despite its simplicity, this stain has stood the test of time.
Even now, over a century later, it remains the most frequently used tissue stain worldwide.
H&E
Deparaffinization
It is the removal of wax is done with_____.
xylene
It is essential to remove the wax completely, otherwise subsequent stages will not be possible.
At least _____changes in xylene are needed to ensure complete removal.
2 to 3
Sections at this stage should appear clear and transparent.
Presence of any patches indicates the presence of wax and sections should be kept longer in the_____.
Deparaffinization
xylene
_______is not miscible with aqueous solutions that are used later in the staining process.
Therefore it is removed with graded alcohol from higher concentration to lower concentration.
Hydration
Xylene
Hydration
First change is made to____________ and finally_____.
absolute alcohol or acetone followed by 90, 70% alcohol
distilled water
After hydration
Sections now should appear…
opaque
Hydration
Presence of any clear areas are indicative of the presence of_____. To remove this, sections should be returned to absolute alcohol and rehydrated.
xylene
(whenever necessary)
In case mercury containing fixatives e.g. Zenker is used, mercury pigments are precipitated on the sections.
Removal of mercury pigments
Removal of mercury pigments
Removal involves treatment with____ solutions which changes mercury to an _____ compound.
This in turn is converted to______ by_____, which is readily soluble in water, the slides are placed in running water to wash out all extraneous chemicals.
iodine
tetrathionate
thiosulphate
Staining Various staining reagents are applied during this hydrated stage.
This involves exposing the tissues to_____, _____ and strong chemicals like_____ and ______. Care must be observed during this stage to avoid detachment of tissues.
hematoxylin, eosin Y
acid alcohol and ammonia
Dehydration and clearing
Dehydration is done using graded alcohols from_____ to _____
70% to absolute alcohol.