Hematoxylin and Eosin Staining Method Flashcards

1
Q

For routine microanatomical examination,_______is the staining method of choice

A

Hematoxylin and Eosin (H&E)

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2
Q

H&E

The two stains were independently introduced in____ and ____, respectively, by____ and ____

A

1865 and 1875

Böhmer and Fischer

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3
Q

In 1876,______ described their use in combination as a tissue staining method for staining different materials with different colours.

A

Wissowzky

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4
Q

is the most important type of bonding that occurs in histologic staining techniques.

A

Ionic bonding

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5
Q

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

A

Ionic bonding

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6
Q

alone is not technically a dye, and will not directly stain tissues.

A

Hematoxylin

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7
Q

Hematoxylin needs to be used in combination with a “_____” – a compound that helps it link to the tissue

A

mordant

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8
Q

The mordant used is typically a metal____, such as aluminium.

A

cation

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9
Q

Hematoxylin in complex with aluminium salts is cationic and acts as a______ dye.

A

basic

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10
Q

Hematoxylin is______ charged and can react with______ charged, basophilic cell components, such as nucleic acids in the nucleus.

These stain____ as a result.

A

positively

negatively

blue

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11
Q

is anionic and acts as an acidic dye.

A

Eosin

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12
Q

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.

A

negatively

positively

acidophilic

pink

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13
Q

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.

A

H&E

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14
Q

Deparaffinization

It is the removal of wax is done with_____.

A

xylene

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15
Q

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.

A

2 to 3

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16
Q

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_____.

A

Deparaffinization

xylene

17
Q

_______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.

A

Hydration

Xylene

18
Q

Hydration

First change is made to____________ and finally_____.

A

absolute alcohol or acetone followed by 90, 70% alcohol

distilled water

19
Q

After hydration

Sections now should appear…

20
Q

Hydration

Presence of any clear areas are indicative of the presence of_____. To remove this, sections should be returned to absolute alcohol and rehydrated.

21
Q

(whenever necessary)

In case mercury containing fixatives e.g. Zenker is used, mercury pigments are precipitated on the sections.

A

Removal of mercury pigments

22
Q

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.

A

iodine

tetrathionate

thiosulphate

23
Q

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.

A

hematoxylin, eosin Y

acid alcohol and ammonia

24
Q

Dehydration and clearing

Dehydration is done using graded alcohols from_____ to _____

A

70% to absolute alcohol.

25
can remove some of the stains so time has to be suitably modified to minimize fading
Dehydrating alcohol
26
Since alcohol is miscible in xylene, it is used for clearing the sections. Any sections from which water has not been completely removed would give a_____ appearance after the first xylene. Such sections should be returned to _______and the process is repeated. Mounting is done after ___ or ____xylene.
milky absolute alcohol 2nd or 3rd
27
From the slide warmer, allow the slide to cool for at least___ minutes. Then proceeding with staining
5
28
Procedure 1. (2 changes, 5 minutes each). 2. (10 dips) 3.(3 changes, 10 dips each) 4. Wash slides in_____ and remove excess______ or impurities using a cotton swab. Remember not to touch the tissue sections. Then rinse the slide well with clean water and drain excess water on paper towels. 5. Stain slide in Hematoxylin for ______minutes. Drain excess stain then rinse with water. 6. Decolorize sections in_____ (2 quick dips only). Rinse slides immediately in water. 3|Page 7. Immerse slides in _____until tissue turns blue (average of 5 minutes). Rinse again in water. Drain excess water on paper towels. 8. Stain slide with Eosin Y for ____minutes. Drain excess stain then rinse well with water. Drain excess water on paper towels. 9. Dip in_____(3 changes, 10 dips each). 10. Lastly, dip slides in____ (3 changes, 10 dips each).
Xylene Alcohol formol 95% ethanol soapy water albumin 5-10 acid alcohol ammonia water 1.5 - 3 95% ethanol xylene