H&E Staining

Question 1

1-3

Answer 1

Xylene 3mins

Question 2

4,5,6

Answer 2

100% ETOH - 2mins and 30secs

100% ETOH - 2mins

95% ETOH - 2mins and 30secs

Question 3

7

Answer 3

Running tap water - 3mins

Question 4

8

Answer 4

Hematoxylin - 5mins

Question 5

9

Answer 5

Running tap water - 2mins

Question 6

10

Answer 6

Acid alcohol - 1min and 30secs

Question 7

11

Answer 7

Bluing agent - 1min and 30secs

Question 8

12

Answer 8

95% ETOH - 1min

Question 9

13

Answer 9

Eosin - 4 mins

Question 10

14,15,16

Answer 10

95% ETOH - 2mins

100% ETOH - 3mins

100% ETOH - 3mins

Question 11

17,18

Answer 11

Xylene - 3mins

Question 12

Purpose of first 3 xylene

Answer 12

Deparaffinization

Question 13

What is not included during frozen section biopsy?

Answer 13

First 3 xylene

Question 14

Purpose of decreasing concentration of ETOH?

Answer 14

Rehydration

Question 15

Purpose of rehydration

Answer 15

To make hematoxylin penetrate better by opening the tissue pores

Question 16

What happens if rehydration is not done appropriately

Answer 16

Hematoxylin will not be absorbed

Question 17

Primary stain

Answer 17

Hematoxylin

Question 18

Purpose of acid alcohol

Answer 18

Differentiator or Decolorizer

Enhance the performance of BLUING agent and remove excess hematoxylin

Question 19

Enhance the performance of BLUING agent and remove excess hematoxylin

Answer 19

Acid alcohol

Question 20

Bluing agent can be:

Answer 20

Scott’s tap water
Ammonia water

Question 21

Scott’s tap water

Answer 21

Sodium bicarbonate NaHCO₃ + Magnesium sulfate MgSO4

Question 22

is a bluing solution used in histology to intensify the staining of cell nuclei and basophilic structures in tissue sections after hematoxylin staining

Answer 22

Scott’s solution

Question 23

Purpose of bluing agent

Answer 23

Enhances primary stain (hematoxylin)

Question 24

Enhances eosin stain

Answer 24

95% ETOH

Question 25

Acidic stain Counter stain Secondary stain Background stain

Answer 25

Eosin

Question 26

Purpose of increasing concentrations of ETOH

Answer 26

Dehydration

Question 27

Purpose of dehydration

Answer 27

Remove water in preparation for mounting with xylene

Question 28

Purpose of last batch of xylene

Answer 28

Increase the refractive index Important for easier mounting

Question 29

is the corner stone of tissue-based diagnosis The process stains thin tissue sections so that pathologists can visualize tissue morphology

Answer 29

Hematoxylin and Eosin (H&E) staining

Question 30

The process uses a hematoxylin dye to stain cell nuclei (and other parts)_____ and an eosin dye to stain other structures____

Answer 30

blue pink or red

Question 31

binds strongly to acids and consequently binds to nuclear DNA and stains nuclei blue. Properly applied, this technique provides exceptional detail of tissue structure and the makeup of the cells. This detail is required for tissue-based diagnosis, particularly in the detection and classification of infection, cancer or metabolic disease

Answer 31

Hematoxylin

Question 32

plays a significant role in tissue-based diagnosis by coloring otherwise transparent tissue sections, and allowing cell structures including the cytoplasm, nucleus, and organelles and extra-cellular components to be clearly visible under the microscope

Answer 32

Routine H&E staining

Question 33

In a histology laboratory, all specimens are initially stained with_____ and additional stains are only ordered if additional information is needed to provide a more detailed analysis.

Answer 33

H&E

Question 34

The term______ is used by pathologists to indicate any area of damage, infection, inflammation, tumor, necrosis or otherwise abnormal tissue.

Answer 34

lesion

Question 35

ROUTINE H&E STAINING in Paraffin Embedded Section Progressive or Regressive?

Answer 35

Regressive Staining

Question 36

Routine H&E staining Fixation: Most fixatives can be used except_____ solutions which inhibit hematoxylin.

Answer 36

osmic acid

Question 37

Differentiate in_______ (________)

Answer 37

1% acid-alcohol 1 ml concentrated HCl to 99 ml of 80% ethyl alcohol

Question 38

Bluing agent Blue in_______ (average of 5 minutes) or ________until the sections appear blue (about 30 seconds).

Answer 38

ammonia water 1% aqueous lithium carbonate

Question 39

Counterstain with ______for 5 minutes. If alcoholic eosin is used, the time can be reduced to 30 seconds or 1 minute

Answer 39

5% aqueous eosin

Question 40

For tissues fixed with_______, the staining time in hematoxylin should be increased slightly while duration of eosin staining should be reduced

Answer 40

mercuric chloride

Question 41

The following staining methods are commonly employed for frozen sections, the choice depending upon the personal preference of the pathologist and the type of tissue section to be stained.

Answer 41

1. Hematoxylin-Eosin method 2. Thionine method 3. Polychrome Methylene Blue method 4. Alcoholic Pinacyanol method

Question 42

H & E staining of Frozen Sections for Rapid Diagnosis Progressive or Regressive?

Answer 42

Progressive Staining