STAINING Flashcards
trans base
The process of applying dyes on the sections
staining
To see and study the architectural pattern of the
tissue, the physical characteristics of the cells,
and the structural relationship of the tissue and
their cells
staining
Nucleus (acidic) attracts what dye
basic dye
cytoplasm (basic) attracts what dye
acidic dye
Direct interaction between the
composition of the tissues and the stain solution being used in the process
Histological Staining
applying dye through a chemical reaction
Histochemical Staining
Combination of immunologic and
histochemical techniques allowing to
look for phenotypic markers onto the
tissues
Immunohistochemical Staining
special procedures being performed in the histopathology lab, meaning, these are rarely performed compared to
Histological staining
Histochemical and Immunohistochemical staining
no other processes involved, sections are stained with simple Aqueous or
Alcoholic Solutions
direct staining
Involves the usage of Mordant in order for the stain to interact or stay on the tissue
indirect staining
“link or bridge” between tissue and
dye; added with dye (lake)
mordant
accelerate the speed of staining
accentuator
Follows a definite sequence and the concentration is ALWAYS increasing
progressive staining
Tissue is first overstained; decolorized
Decreasing
regressive staining
selective removal of excess stain
Differentiation/Decolorization
Forms a soluble salt with the metal so that the latter is dissolved out
Acid Differentiator
Oxidizes the dye to a colorless substance
Oxidizing Differentiator
If the primary stain is basic, the decolorizer is acidic, and vice-versa
Mordant Differentiator
acts as a differentiator for both
basic and acidic dye
alcohol
Stains tissue with a color that is different from that of the stain itself
metachromatic staining
what metachromatic stain is used on mast cells
toluidine blue
what metachromatic stain is used on reticulocytes
cresyl blue
Application of a different color or stain to provide contrast and background staining
counterstaining
stains used in cytoplasmic
○ Eosin Y
○ Picric Acid
○ Lissamine Green
stains used in nuclear
○ Neutral Red
○ Hematoxylin
○ Safranin O
General relationship of tissues and cells
microanatomical staining
structures found in the cytoplasm and nucleus
cytoplasmic staining
stain for bacterial morphology
negative staining
Demonstration of tissue elements by the use of metallic salts (colorless solutions)
metallic impregnation
produce an opaque or black
deposits on the tissue
metallic salts
part of the tissue that had bacteria on it
black deposits
Selective staining of living cells
vital staining
stain for RER
trypan blue
stain for mitochondria
janus green
Done by injecting the dye into any part of animal body
intravital staining
to demonstrate the uterus in the
gravid helminths in parasitology
india ink
Stains living cells immediately after removing from the living body
supravital staining
two types of stains and staining solutions
natural dyes
synthetic dyes
active coloring agent of hematoxylin
hematin
Derived from Hematoxylin campechianium
From the heartwood of a mexican tree
Originally found in Campeche, Mexico.
Most valuable stain used
hematoxylin
types of hematoxylin
Alum Hematoxylin
Iron Hematoxylin
Copper Hematoxylin
Progressive staining
Counterstained with Eosin, Congo Red
Safranin
Alum Hematoxylin
For differential or regressive staining
Iron Hematoxylin
Used for the process of spermatogenesis
Copper Hematoxylin
Derived from female Coccus cacti (Cochineal bug). Widely used a powerful chromatin and nuclear stain
conchineal dyes
“Coal Tar Dyes”; derived from Hydrocarbon benzene
aniline dyes
When treated with Alum, will be termed as
Carmine
dye appears in the tissue
Chromophores
for Neuropathological studies
Carmine w/ Picric Acid
for Glycogen Demonstration, considered as the best Carmine stain
Carmine w/ Aluminum Chloride
Derived from Lichens; a vegetable dye
For staining Elastic Fibers
orcen
gives color to the dye
Chromogens
For regressive staining
Cartilage and cement lines of bones: Blue
Ehrlich’s Hematoxylin
soluble in alcohol but not in water
Neutral dye
base must be Sodium
Acid dye
staining time for PTAH
12-24 hrs
Routine nuclear staining
Exfoliative cytology
Sex chromosomes
Harris Hematoxylin
staining time for coles Hematoxylin
10 mins
what color does PTAH produce for Nuclei, fibrin, muscle striations, myofibrils, fibroglia
blue
based must be HCl
Basic dye
Mucopolysaccharide
Nuclear staining
Mayer’s Hematoxylin
staining time for ehrlichs hematoxylin
15-40 mins
staining time for harris Hematoxylin
5-20 mins
Routine Purposes
Used in sequence with Celestine Blue
Coles Hematoxylin
Demonstrating muscle fibers and
connective tissue
Recommended when the preceding stains contain acid
Weigert’s Hematoxylin Solutions
Demonstrates structure in paraffin,
celloidin, and frozen section
Phosphotungstic Acid Hematoxylin (PTAH)
Cytological stain recommended for
regressive staining of thin sections
Demonstration of chromatin,
chromosomes, nucleoli, centrosomes, and mitochondria
Voluntary muscle striation and myelin
Heidehain’s Hematoxylin
what color does PTAH produce for Collagen, bone, cartilage
Orange-Red or Brownish Red to Deep Red Stain
Staining connective tissue and cytoplasm differentially
eosin
Basic nuclear stain; diagnosis of
Diphtheria; Fresh sputum; Aerogenes bac
methylene blue
MB is heated with fixed alkali/alkali
carbonate
methyl violet
Sub for Thionine in FFT
For Nissl granules
toluidine blue
Amyloid in FFT; Platelets
For Hematology
crystal violet
Counterstaining of epithelial sections
aniline blue
Plasma stain for Acid-fast organism
basic fuchsin
Ascaris eggs and erythrocyte; Bacterial
spore stain
Both decolorizer and counterstain
Usually for Parasitology
malachite green
Contrast stain for Gran’s staining
bismarck brown
Utilized for manufacture of paints
prussian blue
Acid mucopolysaccharide
alcian blue
For Embryos
congo red
For Mitochondria
janus green
Substitute for Carbole Fuchsin
night blue
Used to stain lipids or fats
lysochromes
stain specific for lipids
sudan
For Phospholipids & Neutral Lipids
0.5% solution boiled in 70% ethanol
sudan black
For neutral lipids only
2% Benzoic acid in 70% Alcohol
sudan IV
Fat stain for CNS
sudan III
Should always be distilled
water
Ethyl alcohol or Methyl alcohol
alcohol
10cc added to every 1⁄2 to 1 L of hot
distilled water
aniline water
Holes 5-9 slides
coplin jar
Aqueous solution of 0.5-5%
phenol
To protect the specimen form physical injury
mounting of sections
Holds 5-19 slides
slotted staining dishes
Holds 10-30 slides
metal/glass staining rack/carries
recommended time for mounting
10 years
RI of good mounting medium
1.518
for water-miscible preparations
aqueous media
used when dehydration and clearing
with Xylene cannot be made
glycerin jelly
RI for glycerin jelly
1.47
Farrant’s Medium
gum arabic
RI of gum arabic
1.43
RI for apathy’s medium
1.52
Preparations that have been dehydrated and cleared in Xylene and Toluene
resinous media
RI for canada balsam
1.524
RI for dpx
1.532
RI for xam
1.520
RI for permount
1.518
RI for clarite
1.544
Process of sealing the margins of the coverslip to prevent escape of fluid
ringing
2 parts of paraffin wax mixed with
4-9 parts powdered Colophonium Resin
Kronig Cement