Staining Flashcards
1
Q
to study the architectural pattern of the tissue and physical characteristics and structural relationships
A
staining/dyeing
2
Q
staining is made possible through ___
A
capillary osmosis, solubility, absorption, and adsorption of stains or dyes by tissues
3
Q
PRINCIPLE OF STAINING
Alkaline structures (__) are stained by __ (Eosin cystoplasm is stained __),
**Acidic structures **(nucleus) are stained by ___ (hematoxylin; nucleus is stained __)
A
Alkaline structures (cytoplasm) are stained by acidic stains (Eosin cystoplasm is stained red),
Acidic structures (nucleus) are stained by alkaline/basic stains (hematoxylin; nucleus is stained blue)
4
Q
main function of staining
A
to enhance the visualization of cells and certain cellular component
5
Q
histological staining is for?
A
microanatomical staining, bacterial staining, and specific staining for tissues (muscles, connective, neurologic stains)
6
Q
used to demonstrate the general relationship of tissues and cells with differentiation of nucleus and cytoplasm
A
histological staining
7
Q
provides emphasis for the specification and identification of muscles, connective tissues, and neurologic stains
A
histological staining
8
Q
constituents of tissues are studied thru chemical reactions
A
histochemical staining
9
Q
enumerate examples of histochemical staining and what method is associated with this staining?
A
- Perl’s Prussian Blue
- Periodic Acid Schiff
SPECIFIC STAINING
10
Q
Perl’s Prussian blue stains for __ which demonstrates ___.
A
stains hemoglobin which demonstrates ferric iron in tissues
11
Q
Periodic Acid Schiff stains?
A
carbohydrates
12
Q
this staining allows phenotypic markers to be detected and demonstrated using a wide range of monoclonal, fluorescent labeled or ___
A
immunohistochemical staining
enzyme-labeled antibodies
13
Q
enumerate the methods of staining
A
- specific
- simple/direct
- indirect
- progressive
- regressive
- metachromatic
- counter staining
- vital staining
14
Q
accomplished by controlled, specific chemical reactions designed to give final color (staining) at the site/location of the structure of the substances in the cells or tissues
A
specific staining
15
Q
specific staining is the basis of?
A
histochemistry
16
Q
Perl’s prussian blue would give specificity/enhance and demonstrate the ___
A
hemosiderin granules
17
Q
Periodic acid schiff technique would give specificity/enhance and demonstrate the ___
A
polysaccharides
18
Q
simple/direct staining is the staining of tissue by means of ___ of the dye
A
simple alcoholic/aqueous solution of the dye
19
Q
in this method of staining, the action of the dye is intensified by some other reagent
A
indirect staining
19
Q
give an examples of simple staining
A
methylene blue and eosin
20
Q
why it is called indirect staining?
A
because it still need mordant and accentuator
21
Q
substance which when taken up by the tissue, helps make the in return serving as a link or bridge to make the staining reaction possible
A
mordant
22
Q
mordant combines with a dye forming a colored __ which combines with tissue to form an insoluble ____
A
lake which combines with tissue to form an insoluble tissue-mordant-dye complex
23
Q
an integral part of the staining reaction itself
A
mordant
24
this does not participate but merely increases or heightens the color intensity, selectivity and crispness of the stain
accentuator
25
what is the difference between mordant and accentuator
accentuator do not bind or link the tissue to the dye rather it just increases or heightens the color intensity, selectivity, and crispness of the stain
26
enumerate the examples of a mordant
- Potassium Alum with Hematoxylin in Ehrlich's hematoxylin
- Iron in Weigert's hematoxylin
27
enumerate the examples of accentuator
- Potassium hydroxide (KOH) in Loeffler's Methylene blue
- Phenol in carbo thionine and carbol fuchsin
28
mordant and accentuator is related to what method?
indirect staining method
29
staining is continued in a definite sequence until the desired intensity of coloring of the different tissue elements is attained
progressive staining
30
no washing/differentiation/decolorization in between is required solely relies on the selective affinity of dyes for different cellular elements, what method of staining?`
progressive staining
31
t or f: in regressive staining, any staining is possible as long as no differentiation is done
false, progressive staining
32
in this method, the tissues are overstained and the excess dye is then removed until the desired intensity is obtained
regressive staining
33
principle/s of regressive staining
- overstaining
- washing/differentiation/decolorization
34
the selective removal of excess stain from tissue in regressive staining
washing/differentiation/decolorization
35
enumerate the examples of regressive staining
acid fast and gram staining
36
t or f: progressive staining requires differentiation in a dilute acid alcohol
false, does NOT require
37
which is more hematoxylin concentrated? progressive or regressive?
regressive
38
metachromatic staining is also known as
metachromasia
39
use of the specific dyes that stains tissues with a color that is different from that of the stain color itself
metachromatic staining
40
metachromatic staining is particularly employed for staining of?
cartilage, epithelial mucins, connective tissues, amyloid, mast cell granules
41
metachromatic dye are the basic dyes belonging to ___
Thizine and Triphenylmethane groups
42
enumerate the examples of metachromatic
cresyl blue for reticulocytes
43
application of a different color or stain to provide contrast and background to the staining
counterstaining
44
eosin is an example of counterstaining for the __
cytoplasm
45
selective staining of living cell constituents, demonstrating cytoplasmic structures by the ___ of the dye particle
vital staining - phagocytosis
46
vital stain of reticuloendothelial system
trypan blue
47
demonstration of nuclear structures during vital staining suggests __ of the membrane by the dye, signifying the death of the cell
suggest **permeability**
vital staining
48
true vital (intravital) staining of mitochondria
Janus Green B
49
intravital staining is done by injecting the dye into ____ producing specific coloration of certain cells
intravenous, intraperitoneal, subcutaneous
50
enumerate the examples of intravital staining
lithium, india ink, carmine
51
used to stain living cells immediately after removal from the living body
supravital staining
52
best supravital dye
neutral red
53
metallic impregnation has its greatest application in tissue from ___ and for the demonstration of __
CNS and for the demonstration of reticulin
54
this consists of an opaque black particular precipitate
metallic impregnation
55
most commonly used agent for impregnation
silver nitrate
56
used for demonstration of lipids
osmium tetroxide
57
enumerate the natural dyes
SHOC - Saffron, Hematoxylin, Orcein, Cochineal
58
natural dyes are obtained from?
plants and animals
59
most commonly used for routine histologic studies
hematoxylin
60
A vegetable dye extracted from lichens which are
normally colorless, but when treated with ammonia and
expose to air, produce blue or violet color
orcein
61
used for staining Elastic fibers
orcein
62
An old histologic dye extracted from the female cochineal bug (coccus cacti)
cochineal
63
A plant with orange stigmas yielding a dye
saffron
64
synthetic dyes are also known as __
coal tar dyes
65
synthetic dyes are derived from the ___ and are collectively known as __
hydrocarbon benzene and are collectively known as **aniline dyes**
66
a synthetic dye should consist these two substances, enumerate.
chromophores and auxochromes
67
Substances capable of producing visible colors, also known as “color-bearers” in greek
chromophores
68
Substances which impart to the compound property of electrolytic dissociation, thereby altering the shade of the dye, giving it the property
of forming salts with another compound and ultimately retaining its color, this is also known as "color-increaser"
auxochromes
69
Natural dye that is extracted from the core of heartwood of the tree **Haematoxylon campechianum**
hematoxylin
70
formed by oxidation of Hematoxylin
hematin
71
this is done by exposing the substance to air and
sunlight, thereby oxidizing hematoxylin
ripening
72
The complex of stain and mordant in hematoxylin is called a
“lake”
73
Most commonly used mordant. Gives a **blue lake appearance**. This increase the selectivity for nuclei, especially if acid is added or is used as a differentiating agent
aluminum
74
enumerate the mordants of hematoxylin
aluminum, chromium, iron, copper, tungsten/phosphotungstic acid, ferric salt
75
this causes an intense **blue-black appearance** when used
as a mordant
ferric salt
76
most commonly used eosin
eosin Y
77
A red acid dye used for cytoplasm differentially. Commonly used as a background or contrast stain
because it gives a leasing and useful contrast to nuclear stains such as hematoxylin
eosin
78
H&E STAINING COLOR REACTION
cytoplasm and proteins in edema fluid
pale pink
79
H&E STAINING COLOR REACTION
decalcified bone, osteoid, cartilage
pink
80
H&E STAINING COLOR REACTION
muscle fiber
deep pink
81
H&E STAINING COLOR REACTION
karyosome
dark blue
82
H&E STAINING COLOR REACTION
cartilage
light blue to dark blue
83
H&E STAINING COLOR REACTION
RBC, Eosinophil granules, keratin
bright orange to red
84
H&E STAINING COLOR REACTION
calcium and calcified bone
purplish blue
84
H&E STAINING COLOR REACTION
plasma cells, osteoblast, basophilic cytoplasm
purplish pink
84
H&E STAINING COLOR REACTION
nucleus
blue to blue black
85
specialized fixative used in frozen section which serves to localize antigens and hydrolytic enzymes
formol calcium
86
formol calcium is also used for?
preservation of lipids
87
what makes it different from H&E from the modified H&E technique?
they are similar but the differentiation stage is omitted, it is a progressive form of staining
88
enumerate the 3 stains of papanicolaou stain
- hematoxylin
- OG-6 (orange green)
- eosin azure
89
enumerate the composition of eosin azure
- eosin
- bismarck brown
- lithium carbonate
- PTA
- light green stain (36, 50, 65)
90
91
hematoxylin in papanicolaou is for staining?
nuclear structures
92
OG-6 (orange green) is for staining?
cytoplasm of mature cells
92
eosin azure is for staining?
cytoplasm of immature cells
93
what is omitted in the modified pap's stain?
bismarck brown in the eosin azure
94
Van Gieson's is a mixture of __ for the demonstration of ___
mixture of picric acid and acid fuchsin for the demonstration of connective tissues, mucin and elastic tissue
95
in ___, basic acridine fluorochrome permits discrimination between ____ giving ___ for DNA
acridine orange
discrimination between dead and living cells giving GREEN fluorescence for DNA
96
for calcium salts and phosphate activity
acridine red 3B
97
alcian blue stain ___, it is more specific for ____
stain acid mucopolysaccharides, it is more specific for connective tissue and epithelial mucin
98
enumerate the basic fuchsin
Carbol fuchsin, Coleman's Feulgen, Schiff's, Mallory's fuchsin, Aldehyde fuchsin (Gomori)
99
a plasma stain utilized also for deep staining of acid fast organisms mitochondria, and differentiation of smooth muscles with the use of ___
basic fuchsin
use of picric acid
100
used for staining hemoglobin
benzidine
101
used for staining ___, used for contrast stain for Gram's technique, acid fast, and Papanicolau method
Bizmarck brown
used for staining diptheria
102
used as a chromatin stain for fresh materials in smear preparations
carmine
103
best carmine is?
combined with aluminum chloride to stain glycogen
104
a mordanted dye acting as a basic dye and staining acid substances
Mayer's Carmalum solution
105
recommended for routine staining of fixed sections
celestine blue
106
___ best known as indicator utilized as a stain for ____ in embryos. used for staining ___, ____, and ___ (Krajian's method)
congo red utilized as a stain for AXIS CYLINDERS in embryos. used for staining ELASTIC TISSUES, AMYLOID, AND MYELIN (Krajian's method)
107
a nuclear or chromatic stain used for staining amyloid in frozen sections and platelets
crystal violet
108
gentian violet is a mixture of?
crystal violet, methyl violet, and dextrin
109
used for staining blood to differentiate leukocytes
giemsa
110
composition and purpose of gold sublimate
made up of gold chloride and mercuric chloride, *used for metallic impregnation*
111
the oldest of all stains
gram's and lugol's iodine
112
gram's/lugol's iodine stains __
amyloid, cellulose, starch, carotenes, and glycogen
113
purpose of malachite green
- staining ascaris eggs and erythrocytes
- used as a bacterial spore stain
114
stains chromatin green in the presence of an acid
methyl green
115
purpose of methylene blue
- for plasma cells,
- fresh sputum for malignant cells
- evaluation and differentiation of bacterial organisms
- for diptheria diagnosis
- nervous tissue vital staining
116
methylene violet, coloring nuclei of leukocytes ___ in presence of ___
reddish purple in presence of methylene blue
117
used for observing cell granules and vacuoles of phagocytic cell
neutral red
118
substitute for carbol fuchsin in acid fast staining
night blue
119
for dermatological studies
orcein
120
orcein is an excellent stain for ___ which demonstrates the finest and most delicate _____
for elastic fibers which demonstrates the finest and most delicate fiber skin
121
osmium tetroxide used as a fixative used to stain __
fat
122
normally utilized for the manufacture of paints used as microanatomical contrast stain for demonstration for the circulatory system by injection (intravital staining)
prussian blue
123
___ used with osmic acid to fix and stain blood and granular tissue
rhodamine B
124
used in identification of spirochetes, reticulum, and other fiber stains
silver nitrate
125
recommended for staining Nissl granules or chromophilic bodies; nuclear stain for fixed tissues
toluidine blue
126
toluidine blue is used as a substitute for ___ in fresh tissue sections
thionine
127
used for demonstration of neuroglia in frozen section
victoria blue
128
enumerate the cytoplasmic counterstains: RED
eosin Y, eosin B, phloxine B, rose bengal
129
enumerate the cytoplasmic counterstains: YELLOW
picric acid and Orange G
130
enumerate the cytoplasmic counterstains: GREEN
light green SF and lissamine green
131
enumerate the nuclear counterstains: RED
neutral red, safranin, carmine
132
enumerate the nuclear counterstains: BLUE
methylene blue, tuluidine blue, celestine blue, hematoxylin
133
enumerate the lysochrome or the oil soluble dyes
- Sudan black B
- Sudan IV or Sharlach R
- Sudan III
134
this lysochrome has greater affinity for phospholipids or neutral fats (triglycerides) which is more sensitive coloring agent than other lysosomes
Sudan Black B
135
this lysochrome demonstrates lipids that are resistant to paraffin embedding
Sudan Black B
136
T or F: Sudan IV should be discarded if the brownish black color appeared
false, Sudan Black B
137
this lysochrome is recommended for neutral fats like triglycerides
Sudan IV/Sharlach R
138
T or F: Sudan IV **do not color** phospholipids and fine lipid droplets
true
139
the first Sudan Dye introduced into Histochemistry, fat soluble
Sudan III
140
T or F: Sudan Black B is a good stain for CNS
false, Sudan III
141
this is spread thinly and evenly on a clean grease-free glass slide which is then gently approximated to the end of the ribbon and drawn upwards in a near vertical motion
adhesives
142
enumerate the most commonly used adhesives (8)
- Mayer's egg albumin
- Dried albumin
- 1% gelatin
- Gelatin
- Starch paste
- Plasma
- Poly-L-cysteine
- 3-APES (3-aminopropyltriethoxysilane)
143
144
