STEP 7: STAINING Flashcards
The purified form of a coloring agent or crude dye that is generally applied in an aqueous solution
Histologic stain
The process of giving color to the sections by using aqueous or alcoholic dye solutions
Direct staining
The process whereby the action of the dye is intensified by adding another agent to make the staining reaction possible
Indirect staining
Serves as a link or bridge between the tissue and the dye; it combines with a dye to form a colored “lake”
Mordant
Does not participate in the staining reaction, but merely accelerates the reaction
Accentuator
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 elements is attained
Progressive staining
Tissue is overstained to obliterate the cellular details, and the excess stain is removed or decolorized from unwanted parts of the tissue, until the desired intensity of color is obtained
Regressive staining
Most common method utilized for microanatomical studies of tissues using the regressive staining
Routine Hematoxylin and Eosin staining
Studies the general relationship between tissues and dyes
Microanatomical studies
The selective removal of excess stain from the tissue during regressive staining
Differentiation (decolorization)
The use of specific dyes which differentiate particular substances by staining them with a color that is different from that of the stain itself
Metachromatic stainig
Dyes more effective than methyl violets
Azure or toluidine blue dyes
The process where specific tissue elements are demonstrated, not by stains, but by colorless solutions of metallic salts which are thereby reduced by the tissue, producing an opaque, usually black deposit on the surface of the tissue or bacteria
metallic impregnation
Most valuable metals as metallic impregnating agents
Gold chloride and silver nitrate
Selective staining of living cell constituents, demonstrating cytoplasmic structures by phagocytosis of the dye particle, or by staining of pre-existing cellular components
Vital staining
Staining of pre-existing cellular components
True vital staining
Injecting the dye into any part of the animal body, producing specific coloration of certain cells, particularly those of the reticuloendothelial system
Intravital staining
Method of staining used in microscopy to examine living cells that have been removed from an organism
Supravital staining
Lithium, carmine, and india ink are examples of what stain?
Intraviral stains
NMB, BCB, neutral red, janus green, and trypan blue are examples of what stain?
Supravital stain
Supravital stains for reticulocytes
New methylene blue
Brilliant cresyl blue
Best vital dye
Neutral red
Supravital stain for mitochondria
Janus green
Supravital stain that is toxic to cell
Trypan blue
Application of a different color or stain to provide contrast and background to the staining of the structural components to be demonstrated
Counter staining
3 red cytoplasmic stains
Eosin Y
Eosin B
Phloxine B
3 yellow cytoplasmic stains
Picric acid
Orange G
Rose bengal
2 green cytoplasmic stains
Light green SF
Lissamine green
4 red nuclear stains
Carmine
Hematoxylin
Neutral red
Safranin O
3 blue nuclear stains
Methylene blue
Toluidine blue
Celestine blue
Process whereby various constituents of tissues are studied thru chemical reactions that will permit microscopic localization of a specific tissue substance
Histochemical staining
Widely used in the diagnosis of abnormal cells
Immunohistochemical staining
Staining technique using a wide range of polyclonal or monoclonal, fluorescent labeled or enzyme-labeled antibodies to detect tissue antigens and phenotypic markers under the microscope
Immunohistochemical staining
Hematoxylin is derived from the core or the heartwood of the Mexican tree…
Hematoxylin Campechianum
An active coloring agent formed by the oxidation of hematoxylin, a process known as “ripening”
Hematin
Mordant of Alum hematoxylin
Potassium alum
Alum hematoxylin used for regressive staining
Erlich’s
Ripening agent of Ehrlich’s hematoxylin
Sodium iodate
Alum hematoxylin used for routine nuclear staining in exfoliative cytology
Harris’ hematoxylin
Ripening agent of Harris’ hematoxylin
Mercuric oxide
Ripening agent of Cole’s hematoxylin
Alcoholic iodine
Hematoxylin for regressive and progressive staining
Mayer’s hematoxylin
Ripening agent of mayer’s hematoxylin
Sodium iodate
Hematoxylin for differential or regressive staining; acid-alcohol as a differentiating agent
Iron hematoxylin
Standard iron hematoxylin used in the laboratory
Welgert’s hematoxylin
Ripening agent and mordant of Welgert’s hematoxylin
Ferric chloride
Iron hematoxylin used for regressive staining
Heidenhain’s hematoxylin
Ripening agent and mordant for Heidenhain’s hematoxylin
Ferric ammonium sulfate
Hematoxylin for progressive staining
Ripening agent: KMnO4
Mordant: 1% phosphotungstic acid
Phosphotungstic acid
Hematoxylin used for the study of spermatogenesis
Copper
Cochineal dye is extracted form the cochineal bug…
Coccus cacti
Cochineal dye is treated with alum to produce the dye…
Carmine
Used for neuropathological studies
Cochineal dye combined with picric acid
Best carmine stain; for the demonstration of glycogen
Cochineal dye combined with aluminum chloride
Vegetable dye from certain lichens which are normally colorless, but turn blue or violet when treated with ammonia and exposed to air
Orcein
Color of orcein when treated with ammonia and exposed to air
Blue or violet
Mainly used for staining elastic fibers
Orcein
Stain extracted form flower of crocus sativus
Saffron
Derived from the hydro-carbon benzene
Synthetic dyes (collectively known Aniline dyes)
Coloring property of synthetic dyes is attributed to the…
Chromophore
Active coloring substance is found in the acid component, and the inactive base is the sodium salt of a sulfonate of rosaniline
Acid dyes
Active coloring substance is found in a basic component that combines with the acid radical
Basic dyes
Formed by combining aqueous solutions of acid and basic dyes
Neutral dyes
Oldest stain; Stains amyloid for microscopic study of starch granules
Iodine
The process where hematoxylin is oxidized to hematin
Ripening
Karyosome color stained with hematoxylin
Dark blue
Nucleus color stained with hematoxylin
Blue
Cytoplasm color stained with hematoxylin
Pink
Most valuable stain; used as a counterstain
Eosin
Eosin with blue-deep red color
Eosin B
Eosin with yellow-green color
Eosin Y
Common basic nuclear stain; valuable for plasma cells
Methylene blue
Metachromatic dye; for leukocytes
Methylene violet
Nuclear stain substitute for thionine for fresh frozen tissue; Nissl/Tigroid granules and chromophilic bodies
Toluidine blue
For amyloid, fungi, platelets in blood
Crystal violet
Counterstain for epithelial cells
Aniline blue
Is a plasma stain; deep staining for acid fast organisms
Basic fuschin
Mixture of picric acid and acid fuschin; for the demonstration of connective tissue; SIMPLEST method of differential staining of collagen
Van gieson
Used for staining blood to differentiate leukocytes
Giemsa
Resistant to strong acids; Recommended for routine staining of fixed sections; ALTERNATIVE to Iron Hematoxylin nuclear stain
Celestine blue
Stains Ascaris eggs, RBCs, Bacterial spore (BOTH A DECOLORIZER AND A COUNTERSTAIN)
Malachite green
Stains chromatin green; It gives false positive reactions with certain secretions such as mucin
Methyl green
Most reliable and specific histochemical staining technique for DNA
Feulgen’s stain
Contrast stain for Gram’s stain, In Acid Fast, Papanicolau method & Diphtheria organism
Bismarck Brown
Calcium: BLACK
Von Kossa Silver Nitrate
Colored Salt of Ferric ferrocyanide, normally used for the manufacture of paints
Prussian Blue
Excellent stain for elastic fiber; _______ + Ammonia —exposed to air→Blue or Violet color
Orcein
For the demonstration of connective tissue (fixative and stain)
Picric acid
Used as a chromatin stain for fresh materials in smear preparations
Carmine
Stains mucopolysaccharide; more specific for connective tissue and epithelial mucins
Alcian Blue
Cell granules & vacuole phagocytic cells
Neutral Red
Best known as an indicator; Utilized as a stain for axis cylinder in embryos; (4% aqueous solution: elastic tissues, amyloid, myelin)
Congo Red
For demonstrating mitochondria during intravital staining
Janus green B
Used for demonstration of neuroglia in frozen sections
Victoria Blue
Is used as a substitute for Carbol Fuschin
Night Blue
DNA: green to blue green; RNA: rose red
Methyl Green Pyronin
Demonstrates deposits of calcium salts and possible sites of phosphatase activities
Acridine Red 3B
Permits discrimination between dead and living cells (DNA: green fluorescence; RNA: red fluorescence)
Acridine Orange
Stains blood and glandular tissues
Rhodamine B
Used for staining haemoglobin
Benzidine
Phospholipids
Sudan Black
Triglycerides and Neutral lipids (deep red)
Sudan IV/Scharlach R
Fats (orange)
Sudan III
Used in Spirochetes reticulum and fiber stains
Silver nitrate
Connective tissues
Azocarmine
Histochemical stain used for the demonstration of carbohydrates (GLYCOGEN)
Periodic Acid Schiff (PAS)
Used for differential staining of pancreatic islets of Langerhans
Aldehyde Fuschin Stain (Gomori)
Staining for muscles and bones; Astrocytes
Mallory’s Phosphotungstic Acid Hematoxylin (PTAH)
Muscle demonstration
Lissamine Fast Red Tartrazine Method
Used for fats
Osmic Acid/Osmium Tetroxide stain
Spirochetes
Levaditi’s method
Melanin (Silver Modification): BLACK
Masson Fontana Technique
Diagnostic for Bile pigments
Gmelin’s Test
Hemosiderin (iron-containing pigment of hemoglobin)
Perl’s Prussian Blue
For Hemoglobin
Benzidine method
Astrocyte
Cajal’s Gold Sublimate
Neurons, Axons, Neurofibrils
Bielschowsky’s Technique
Normal Myelin Sheaths
Weigert-Pal Technique
Copper
Lindquist’s Modified Rhodamine
Bacteria
Gram-Twort
Bacteria, Nocardia, Actinomyces
Brown and Brenn (B&B)
Helicobacter
Cresyl Violet Acetate
Legionella pneumophilia
Dieterle
Spirochetes
Warthin-Starry/Levaditi’s methods
Fungi
Grocott Methamine Silver (GMS)
Hepatitis B Surface antigen
Orcein method
Reticular Fibers
Gordon and Sweet’s method
Enumerate 4 natural dyes
Hematoxylin
Cochineal dye
Orcein
Saffron
Dyes from plants and animals; previously for dyeing of wool or cotton
Natural dyes
Hematin formed from exposing the substance to air/sunlight; 3-4 months
Natural hematin
Hematin from chemical oxidation
Artificial hematin