H&E Staining

Question 1

basophilic

Answer 1

substances that attract basic dyes, blue hematoxylin, ex: nuclei, ribosomes

Question 2

acidophilic

Answer 2

substances that attract acidic dyes, pink eosin, ex: proteins, cytoplasm

Question 3

basic dye

Answer 3

charge on the dye ion is positive, also known as cationic

ex: crystal violet, safranin

Question 4

acidic dye

Answer 4

charge on the dye ion is negative, also known as anionic,

ex: orange G, picric acid

Question 5

mordant

Answer 5

a reagent, often a metal (aluminum, tungsten, iron, chromium) used to link the stain to the tissue

Question 6

lake

Answer 6

the combination of a dye and a mordant

Question 7

hematein

Answer 7

oxidation product of hematoxylin, a weak anionic dye

Question 8

differentiation

Answer 8

process by which excess stain is removed from a tissue so that only the desired element is left stained so it can easily be seen against a clear or counterstained background

Question 9

progressive staining

Answer 9

the reaction is stopped once the desired intensity is achieved. The slide is then rinsed and blued to increase the intensity of the nuclear stain. usually for special stains

Question 10

regressive staining

Answer 10

tissue is overstained and then decolorized with an acidic solution until only the desired element is left stained, this is typically how mordant dyes are used. typically used for hematoxylin staining

Question 11

Delafield

Answer 11

naturally ripened by exposing to oxygen, aluminum mordant, regressive staining

Question 12

Ehrlich

Answer 12

naturally ripened by exposing to oxygen, aluminum mordant, regressive staining

Question 13

Harris

Answer 13

chemically oxidized with sodium iodate, aluminum mordant, progressive staining

Question 14

Mayer

Answer 14

chemically oxidized with sodium iodate, aluminum mordant, does not contain alcohol, progressive staining

Question 15

Gill

Answer 15

chemically oxidized with sodium iodate, aluminum mordant, progressive staining, only hematoxylin that will stain mucin, esp in goblet cells

Question 16

Weigert

Answer 16

ferric chloride (iron) is the oxidizer and mordant, not used in routine H&E, resists acidic solutions

Question 17

hematin

Answer 17

formalin pigment

Question 18

chromogen

Answer 18

benzene derivatives that contain chromogens

Question 19

chromophore

Answer 19

the chemical group that confers color in dyes

Question 20

auxochrome

Answer 20

ionizing groups that enable dye to link firmly to tissue

Question 21

MGP technique (methyl green-pyronin)

Answer 21

stains RNA rose using pyronin and DNA green with methyl green

Question 22

giemsa

Answer 22

polychrome stain; a compound dye made up of different colors that should be pH 6.4 to 6.9

Question 23

Feulgen reaction

Answer 23

demonstrates DNA but not RNA, shows red nuclei only

Question 24

Bouin solution

Answer 24

hydrolyzes nuclei during fixation, doesn’t work with Feulgen reaction

Question 25

acetic acid

Answer 25

makes nuclear staining more selective

Question 26

Romanowsky-type stains

Answer 26

a combination of the basic dye methylene blue and the acidic dye eosin commonly used to differentiate leukocytes (white blood cells)

Question 27

refractive index

Answer 27

should be near that of the tissue

Question 28

refractive index

Answer 28

should be near that of the tissue, tissue appears more transparent as the media's refractive index approaches that pf the tissue

Question 29

cane sugar

Answer 29

can be added to aqueous mounting media to prevent diffusion of basic aniline dyes

Question 30

cornflaking

Answer 30

tissue dries out before coverslipping and appears burnt or singed

Question 31

pale pink cytoplasm in H&E

Answer 31

pH problem with the eosin; due to incomplete removal of bluing agent (too low pH) or too little acid (too high pH)

Question 32

reddish brown nuclei in H&E

Answer 32

over ripened hematoxylin

Question 33

uneven staining

Answer 33

paraffin was not completely removed

Question 34

milky water after hydrating alcohols

Answer 34

xylene has been carried over, change the alcohols to correct this

Question 35

slide dries out before coverslipping

Answer 35

glossy black nuclei, cornflaking, and brown stippling may occur

Question 36

dark nuclei and blue cytoplasm in H&E

Answer 36

inadequate differentiation

Question 37

blue-black precipitate in H&E

Answer 37

filter the hematoxylin to remove aluminum-hematein crystals

Question 38

incomplete dehydration

Answer 38

microscopic water bubbles trapped under the coverslip

Question 39

microscopic water bubbles trapped under the coverslip

Answer 39

incomplete dehydration

Question 40

some parts of tissue can't be brought into focus

Answer 40

mounting media on top of the coverslip

Question 41

hazy blue nuclei

Answer 41

too much heat during processing

Question 42

undo H&E

Answer 42

use acid alcohol

Question 43

last dehydrating alcohol is very pink

Answer 43

contains water which carried over eosin

Question 44

poor Giemsa staining

Answer 44

change the pH of the solution