STAINING Flashcards
process of applying dyes on the sections to see and study the architectural pattern of the tissue and physical characteristics of the cells
staining
purpose of staining
- to make the tissue and cell become more visible
- we can easily identify morphological changes in the tissue/cell
- performed to establish the presence of disease
higher affinity with basic dye
acidic
higher affinity with acidic dyes
basic
tissue constituents are demonstrated in sections by direct interaction with dye or staining solution
histologic stain
producing coloration of the active tissue component
histologic staining
employed to demonstrate the general relationship of tissues and cells with differentiation of nucleus and cytoplasm
histologic stains
examples of histologic stains
- microanatomic stains
- bacterial stains
- specific tissue stains
examples of microanatomic stains
- hematoxylin and eosin
- Masson’s Trichome
- periodic acid schiff
what does hematoxylin stain and what color
it stains nucleus blue
counterstain of hematoxylin
eosin
what does eosin stain and what color
it stains cytoplasm pink
microanatomic stain used for muscle fibers
Masson’s Trichome
color of Masson’s Trichome
red
microanatomic stain used for carbohydrate structures
Periodic Acid-Schiff
color of PAS
magenta
examples of bacterial stains
- gram stain
- Ziehl-Neelsen (Acid-Fast) stain
- Giemsa stain
color of Ziehl-Neelsen (Acid-Fast) stain
red
counter stain of Ziehl-Neelsen (Acid-Fast) stain
methylene blue
primary stain of Ziehl-Neelsen (Acid-Fast) stain
Carbon Fucshin
to heat fat to be easily penetrated by primary stain
Ziehl-Neelsen (Acid-Fast) stain
to differentiate blood cells and parasites by simple staining techniques
Giemsa stain
examples of muscle stains
- Gomori Trichome stain
- ATPase stain
types of specific tissue stains
- muscle stains
- connective tissue stains
- neurologic stains
muscle stain that differentiate muscle fibers from connective tissue
Gomori Trichome stain
combination of different tissues
Gomori Trichome stain
color of muscle fiber and CT in muscle stains
Muscle fibers: red
CT: blue/green
muscle stain for enzymatic reaction
ATPase stain
types of CT stains
- Van Gieson’s stain
- Verhoeff’s Elastic Stain
- Mallory’s Phosphotungstic Acid hematoxylin (PTAH) stain
color of collagen fibers and muscle fibers for Van Gieson’s stain
CF- red
MF - yellow
CT stain for staining elastic fibers
Verhoeff’s Elastic stain
color of elastic fibers for Verhoeff’s Elastic stain
black
color of collagen fibers and muscle fibers for Mallory’s Phosphotungstic Acid Hematoxylin (PTAH) stain
collagen - blue
muscle fibers - red orange
types of neurologic stains
- Luxol Fast blue
- Cresyl Violet (Nissl stain)
- Bielschowsky Silver stain
neurologic stain for myelin sheath
Luxol Fast blue
for staining Nissl bodies
Cresyl Violet (Nissl stain)
stain for studying neurodegenerative substances
Bielschowsky Silver stain
color of Bielschowsky’s Silver stain
Black
tisseu constituents are studied through chemical reactions that will permit microscopic localization of a specific tissue substance
histochemical staining
histochemical staining for hemoglobin
Perl’s Prussian blue
histochemical staining for carbohydrates
Periodic Acid Shiff
part of hemoglobin that is stained
iron/ ferric component of hemoglobin
positive color for iron
blue
ferric iron is converted to
ferric ferrocyanide
detect the activity of alkaline phosphate for bone and liver conditions
alkaline phosphate stain
organs utilized in alkaline phosphate stain
- bone, liver, intestine, and placenta
demonstarate activity of acid phosphate for bone and prostate gland
acid phosphatase
combination of immunologic and histochemical techniques that allow phenotypic markers to be detected and demonstrated
immunohistochemistry
serves as a link or bridge between the tissue and the dye
mordant
not part of the chemical reaction but merely accelerates or hasten the speed of staining reaction
accentuator
3 applications of mordant
- applied before staining
- added as part of the staining solution
- added to dye sltn itself
process fo giving color to the sections by using aqueous or alcoholic dyes; no mordant is used
direct staining
staining reaction is intensified by adding another agent or mordant; mordant is used
indirect staining
process whereby tissue elements are stained in a definite sequence, and the staining solution is applied for specific periods of time or until the desired intensity of coloring of the different tissue element is attained
progressive staining
process of selectively removing excess stain from the tissue
differentiator/decolorizer
tissue is first over stained to obliterate cellular details, and excess stain is removed or decolorized from unwanted parts of the tissue, until the desired intensity of color is obtained
regressive staining
differentiator/ decolorizer is not needed
progressive
differentiator/ decolorizer is needed
regressive
substance is stained with a color that is the same from that of the dye
orthochromic staining
use of specific dye which differentiate particular substance by staining them with a color that is different from that of the stain itself
metachromatic staining
employed for staining cartilage, connective tissue, epithelial mucin, mast cell granules and amyloid
metachromatic staining
color that is different from that of the stain itself
metachromatia
3 methods fo intravital staining
- intravenous
- intraperitonial
- subcutaneous
performed by injecting in the living animal’s body in the blood stream in vivo
vital staining
selective staining of living cell constituents
vital staining
doen by injecting the dye into part of the animal body
intravital staining
example of intravital staining
- lithium
- carmine
- India ink
done by staining living cells immediately after removal from the living body
supravital staining
e.g. of supravital staining
- Neutral red
- Janus green
- Trypan Blue
- Nile blue
- Thionine
- Toluidine blue
trace cell population and their movement within the body
lithium
lysosomes in living organisms
carmine
for staining macrophage
India dye
for staining lysosomes
neutral red
for staining mitochondria
Janus green
for staining dead cells separated from living cells and for cell viability
Tryptan Blue
for staining fat/lipid droplet
Nile Blue
for staining Nissl bodies in neirons
Thionine
for staining mast cells
Toluidine blue
process where specifci tissue elements are demonstrated, not by stains, but by colorless solutions of metallic saltsa which are thereby reduced by the tissue, producing opaque, usually black deposit on the surface of the tissue
matallic impregnation
agent will not be absorbed by the tissue, instead, it is on the surface
metallic impregnation
example of metallic impregnation
ammoniacal silver
reduced by melanin and intestinal glands
ammoniacal silver
obtained from plants and animals previously utilized for dying of wool and cotton
natural dyes
where is hematoxylin derived from
Haematoxylum campechianum
most valuable staining reagent used in cytology
hematoxylin
active coloring reagent/form of hematoxylin
hematin
hematoxylin is usually combined with:
Aluminum, iron, chromium, copper, and salts
oxidation process of hematoxylin
ripening
2 types of ripening
- natural
- artificial
type of ripening wherein hematoxylin is exposed in sunlight or air for 3-4 hours
natural
type of ripening wherein ripening is accelerated by adding oxidizing agents
artificial
examples of oxidizing agents
- H2O2
- mercuric oxide
other name for cochineal dyes
carmine
cochnieal dyes are old histologic dye extracted from the female Cochineal bug called
Coccus cacti
treated with aluminum to produce the dye carmine
Cochineal dye
a powerful chromatin and nuclear stain for fresh material and smear preparation
Cochineal dye
2 solution of cochineal dye
- Picrocarmine
- Best Carmine
composition of picrocarmine
picric acid + cochineal dye/carmine
composition of best carmine
aluminum chloride + cochineal dye/carmine
cochineal dye that is for neuropathologic samples
picrocarmine
cochineal dye for glycogen
Best Carmine
vegetable dye from certain lichens which are normally colorless, but which, when treated with ammonia and exposed to alkali to produce blue or violet color
orcein
used for staining elastic fibers
orcein
used as spice from a stigma of the plant
safron
flower from which safron is harvested from
Crocus sativus
artificial dye is also known as
Coal Tar dyes
derived from the hydrocarbon benzene and are collectively known as Aniline dye
artificial dye
artificial dyes are derived from the hydrocarbon benzene and are collectively known as
aniline dye
composition of artificial dye
- chromophore
- auxochrome
color bearer/coloring property composition of Artificial dye
chromophore
increaser/dyeing property of artificial dye
auxochrome
composition of chromogen
benzene and chromophore
sltn that can impart color temporarily
chromogen
gives dyeing property of dye making it permanent
auaxochrome
active coloring agent is found in the acid acomponent and the inactive base
acid dyes
example of acid dye
picric acid
active coloring substance is found in a basic component that combines with acid radical
basic dyes
example of basic dye
methylene blue
formed by combining aq solutions of acid and basic dyes, capable of staining cytoplasm and nucleus simultaneously and differentially
neutral dyes
counter stain for cytoplasmic staining which is red in color
- Eosin Y
- Eosin B
- Phloxine B
counter stain for cytoplasmic staining which is yellow in color
- Picric acid
- Orange G
- Rose Bengal
counter stain for cytoplasmic staining which is green in color
- Light Green SF
- Lissamine green
counter stain for Nuclear staining which is color red
- Neutral Red
- Safranin O
- Carmine
- Hematoxylin
counter stain for Nuclear staining which is color blue
- methylene blue
- toluidine blue
- celestine blue