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*Special stains*
SPECIAL STAINS
| Question | Answer |
|---|---|
| Detects presence of acid-fast mycobacteria in tissue sections | Kinyoun acid-fast stain |
| lipoid capsule of acid-fast organism takes up carbol-fuchsin and resists decolorization w/dilute mineral acid | Kinyoun acid-fast stain |
| is more soluble in the lipids of the cell wall than in acid-alcohol, is readily removed from bacteria that lack waxy capsule | Carbol-fuchsin |
| Specific way of identifying mycobacteria | Carbol-fuchsin method |
| Although 10% NBF is preferred, others m/b used, avoid carnoy | Kinyoun acid-fast stain |
| 4 to 5 microns | Kinyoun acid-fast stain |
| control- tissue containing acid-fast organisms | Kinyoun acid-fast stain |
| use millipore water in water bath | Kinyoun acid-fast stain |
| negative control from same day work load must be run | Kinyoun acid-fast stain |
| a section from a block of uterus provides good negative control | Kinyoun acid-fast stain |
| overcountestaining w/methelyne blue will amsk any organisms present | Kinyoun acid-fast stain |
| if section is overstained take it back to the acid-alcohol to remove methylene blue, wash with water and repeat counterstaining step | Kinyoun acid-fast stain |
| If section is allowed to dry after the carbol fuchsin, a compound that resist decolorization will be formed. repeat attempts to remove compound results in complete decolorization | Kinyoun acid-fast stain |
| fixatio in Carnoy solution will make acid-fast organisms non-acid-fast | Kinyoun acid-fast stain |
| demonstrate acid mucopolysaccharides | Alcian blue pH 2.5 |
| alcian blue is a copper phthalocyanin basic dye that is water soluble and colored blue bcuz ofits copper content | Alcian blue pH 2.5 |
| forms salt linkages with the acid groups of acid mucopolysacchardes | Alcian blue |
| when used in 3% acetic acid soluton (pH 2.5), alcian blue stains both sulfated and carboxylated acid mucopolysaccharides and sulfated and carboxylated sialomucins (glycogen) | Alcian blue |
| Fixation 10%NBF or bouin | Alcian blue pH 2.5 |
| 4 to 5 microns | Alcian blue pH 2.5 |
| control section of small intestine, appendix or colon (S.A.C) | Alcian blue pH 2.5 |
| Acid-fast bacteria- bright red | Kinyoun acid-fast stain |
| Background - Light blue | Kinyoun acid-fast stain |
| weakly acidic sulfated mucosubstances, hyaluronic acid and sialomucins - dark blue | Alcian blue pH 2.5 |
| background - pink to red | Alcian blue pH 2.5 |
| demonstrate sulfated mucosubstances | Alcian blue pH 1.0 |
| when used in a 0.1N hydrochloric acid solution (pH1.0) alcian blue stains only suflated acid mucopolysacchareds and sulfated sialomucins (glycogen) | Alcian blue pH 1.0 |
| acid mucopolysaccharrides and sialomucins that are carboxylated only will not be stained | Alcian blue pH 1.0 |
| Fixation 10% NBF or Bouin | Alcian blue pH 1.0 |
| 4 to 5 microns | Alcian blue pH 1.0 |
| control - section of small intestine, appendix or colon (S.A.C.) | Alcian blue pH 1.0 |
| sulfated mucosubstances - Pale blue | Alcian blue pH 1.0 |
| Background - pink to red | Alcian blue pH 1.0 |
| differentiate epithelia and connective tissue mucins | Alcian blue w/hyaluronidase |
| staining will disappear or be dramtically reduced when tissue sections containing hyaluronic acid, chondroitin sulfate A or chondroitin sulfate C (CT "mucin") are digested w/ testicular hyaluronidase | Alcian blue w/hyaluronidase |
| Fixation 10% NBF | Alcian blue w/hyaluronidase |
| 4 to 5 microns | Alcian blue w/hyaluronidase |
| label slides one with w/o digestions one w/digestion | Alcian blue w/hyaluronidase |
| control - umbilical cord (with and without). small bowel, appendix or colon for second control to demonstrate epithelial mucins | Alcian blue w/hyaluronidase |
| w/o digestion, acid mucopolysaccharides and sialomucins - deep blue | Alcian blue w/hyaluronidase |
| w/digestion, mucosubstances containing hyaluronic acid and chondroitin sulfate A and C - marked loss staining | Alcian blue w/hyaluronidase |
| differentiate between neutral and acidic mucosubstances | Alcian blue PAS hematoxylin |
| acidic mucosubstances are stained w/alcian blue and neutral mucosubstances are stained by the PAS | Alcian blue PAS hematoxylin |
| Fixation 10% NBF or Zenker solution | Alcian blue PAS hematoxylin |
| 4 to 5 microns | Alcian blue PAS hematoxylin |
| control - kidney or mucin control, depends on the diagnostic tissue to be stained | Alcian blue PAS hematoxylin |
| Exclusively acid mucosubstances - blue, neutral polysaccharides - magenta | Alcian blue PAS hematoxylin |
| Certain substances will be colored by both PAS and alcian blue - purple | Alcian blue PAS hematoxylin |
| demonstrate carboxylated and sulfated mucopolysaccharides and glycoproteins | Muller-mowry colloidal iron |
| Colloidan ferric ions are, at a low pH, absored by carboxylated and sulfated mucosubstances | Muller-mowry colloidal iron |
| Fixation 10% NBF, carnoy or alcoholic formalin, (AVOID CHROMATE FIXATIVES) | Muller-mowry colloidal iron |
| 4 to 5 microns | Muller-mowry colloidal iron |
| control- section of small bowel, appendix or colon (S.A.C) | Muller-mowry colloidal iron |
| acid mucopolysaccharides and sialomucins - deep blue | Muller-mowry colloidal iron |
| nuclei - pink/red | Muller-mowry colloidal iron |
| cytoplasm - pik | Muller-mowry colloidal iron |
| Strongly acidic mucins that do not stain w/alcian blue also do not stain w/colloidal iron | Muller-mowry colloidal iron |
| PAS stain can be used as a counter stain for this procedure | Muller-mowry colloidal iron |
| PAS-positive material will be magenta, mixtures of neutral and acidic mucosubstances will be purple and acidic mucosubstances will be blue | Muller-mowry colloidal iron |
| gives more intense color than alcian blue, colloidal iron is not consideres specific for acid mucopolysaccharides | Muller-mowry colloidal iron |
| Hyaluronidase digestion can be used w/this procedure | Muller-mowry colloidal iron |
| excellent for demonstration of cryptococcus neoformans | Muller-mowry colloidal iron |
| some background stain maby be seen due to the presence of CT mucin | Muller-mowry colloidal iron |
| if strong background staining is noted, fresh solutions should be prepared and stain should be repeated. | Muller-mowry colloidal iron |
| demonstrate amyloid in tissue | Alkaline congo red |
| Green birefringence following congo red stain, is considered most specific technique for demonstration of amyloid | Alkaline congo red |
| false-positive results may be obtained | Alkaline congo red |
| derives from benzidine, can react w/cellulose | Congo red |
| resembles cellulose in its chemical reaction | Amyloid |
| pretreatment w/alkali aids in the release of native internal hydrogen bonds between adjacent protein chains, as a result mre potential sites are available for dye binding | Alkaline congo red |
| is a linear molecule, this allows azo and amine groups of the dye to form hydrogen bonds w/hydroxyl radicals of the amyloid | Congo red |
| Fixation - Alcohol or Carnoy preferred, 10%NBF, Bouin, Zenker may be used, | Alkaline congo red |
| Prolonged storage in 10% formalin will cause gradual decrease in staining intensity | Alkaline congo red |
| cut 8 to 10 microns | Alkaline congo red |
| if sections that are not between 8 to 10 microns, may not show green birefringence | Alkaline congo red |
| control - section w/amyloid | Alkaline congo red |
| don not keep too may control sections cut, as the staining intensity has been reported to decrease with age of the section | Alkaline congo red |
| massive presumably long-standing deposits give less intense histochemical reaction than small, newly formed deposits | Alkaline congo red |
| Amyloid - deep pink to red | Alkaline congo red |
| elastic tissue - pale pink | Alkaline congo red |
| Nuclei - blue | Alkaline congo red |
| following congo red, bright apple-green birefringence exhibited under polarizing light is specific for amyloid | Alkaline congo red |
| false-positive birefringence is caused by excess dye retained in the tissue | Alkaline congo red |
| sodium chloride and high alcohol content present in dye, tend to depress dye ionization and acid-base type staining, results in a stained sectin w/a clean background | Alkaline congo red |
| saturation of solutions is very important, follow instructions carefully | Alkaline congo red |
| is an intrinsic property of the amyloid fibril-congo red complex and is a function of the parallel alignment of dye molecules and amyloid fibrils | Green birefringence |
| section too thin, show red faint color | Alkaline congo red |
| section too thick, show yellow birefringence | Alkaline congo red |
| rapid screening for amyloid | Crystal violet |
| "metachromatic" staining of amyloid is due to the mucopolysaccharide content | Crystal violet |
| amyloid will induce only weak metachromasia w/thionine and toluidine blue | Crystal violet |
| mixtures of basic dye | Crystal violet and Methyl violet |
| Crystal violet and Methyl violet are mixtures of basic dye, amyloid selectively reacts with one of the dye components | Crystal violet |
| the addition of acid to the stain solutions prevents overstaining of the cytoplasmic components | Crystal violet |
| Fixative - 10% NBF or Alcohol | Crystal violet |
| 10 to 12 microns | Crystal violet |
| control - section containing amyloid | Crystal violet |
| Amyloid - purplish violet | Crystal violet |
| Other tissue elements - blue | Crystal violet |
| Bleeding) diffusion into surrounding mounting medium, of basic aniline dyes tends to occur w/aqueous mounting media | Crystal violet |
| stained sections may be allowed to air dry completely then dip in xylene and mount w/synthetic resin | Crystal violet |
| minimum of 5 hours staining time in working crystal violet solution, w/overnight stain providing the ultimate in staining of amyloid w/the crystal violet method | Crystal violet |
| good method for amyloid not as specific as congo red w/polirization | Thioflavine T |
| is a fluorescent dye that attaches to amyloid and requires no differentiation. Background nuclear fluorescence is suprese by staining w/aluminum hematoxylin | Thioflavine T |
| Fixation - 10% NBF | Thioflavine T |
| 6 to 10 microns | Thioflavine T |
| Control - section containing amyloid | Thioflavine T |
| Amyloid- Fluoresces yellow to yellow-green | Thioflavine T |
| Thioflavine T at acid pH increases selectivity of the dye for amyloid. A pH of 1.4 is recommended | Thioflavine T |
| if prefered section must be mounted from water using Apathy mounting media | Thioflavine T |
| lipid granules, jusxtaglomerular granules and mast cells may give a yellow fluorescence but should be differentiated easily from amyliod | Thioflavine T |
| Stains epithelial mucins in tissue | Mayer mucicarmine |
| emperical stain | Mayer mucicarmine |
| aluminum forms a chelation complex with the carmine, resulting compound has a net positive charge and attaches to the acid group of the mucin | Mayer mucicarmine |
| Fixation 10% NBF | Mayer mucicarmine |
| Control - colon, small intestine, appendix (CASI) | Mayer mucicarmine |
| 4 to 5 microns | Mayer mucicarmine |
| Mucin - deep rose/red | Mayer mucicarmine |
| Capsule of cryptococcus - deep rose to red | Mayer mucicarmine |
| Nuclei - black | Mayer mucicarmine |
| other tissue elements - blue o yellow | Mayer mucicarmine |
| carminophillic properties will be obscured if section is overstained with hematoxylin or metail yellow | Mayer mucicarmine |
| anhydrous aluminum chloride should be under a hood. it reacts w/atmospheric moisture and water to give off vapors of hydrogen chloride | Mayer mucicarmine |
| term used to describe intracellular secrections of a variety of cells | Mucin |
| stain with basic dye, metachromatic, precipitated by acetic acid (except gastric mucin) soluble in alkaline solutions | properties of mucin |
| appear to be microscopically similar, they differ in composition | intracellular secretions |
| demonstrate polysaccharides, neutral mucosubstances and basement membranes | PAS reaction |
| oxidation of certain tissue elements to aldehydes by periodic acid. | PAS reaction |
| reactive group is the 1,2 glycol group, other groups are also oxidized | PAS reaction |
| treat basic fuchsin w/sulfurous acid | preparing schiffs reagent |
| reduction casues loss of quinoid structures and masking of the chromophores | PAS reaction |
| colorless compound | leucofuchsin |
| following schiffs reagent, washing in running water causes loss of the bound sulfurous acid group attached at teh centra carbon atom, restoration of quinoid structure in the dye bound tby the aldehyde and the visualization of the typical shiff color | PAS reaction |
| Metabisulfate rinses are used to remove excess schiff reagent and prevent false colorization of the tissue elements due to oxidation of adsorbed reagent | PAS reaction |
| Fixation - 10% NBF or Bouin. blood smears should be fixed in methyl alcohol for 10 to 15 minutes | PAS reaction |
| 4 to 5 microns | PAS reaction |
| Control - section of kidney (most sensitive control). | PAS reaction |
| if demonstrating glycogen use liver containing glycogen or a section of cervix(include both endo and ecto cervix) | PAS reaction |
| Glycogen, neutral mucosubstances, certai epi sulfomucins and sialomucins, colloid material of thyroid, pars intermedia of pituitary, basement membranes, fungal walls | All show a positive PAS raction (Bright rose) |
| schiff staining after periodate does not demonstrate presence of carbohydrate residues | PAS reaction |
| abscence of staining doesnt mean tht carb residues are absent | PAS reaction |
| number of available 1,2 glycol groups, reactivity of schiff reagent w/reaction product, structure of polymer, exact procedural reaction conditions | intensitity of staining in routine PAS reaction is due to the folling factors |
| fast green counterstain m/b used instead of hematoxylin. when stain is used to demonstrate fungus | PAS reaction |
| sulfite rinse essential to remove any uncombined leucofuchsin following exposure to schiff's reagent | PAS reaction |
| highly chlorinated water is capable of oxidation. if section is transferred directly into tap water, any adsorbed schiff reagent m/b reoxidized to basic fuchsin,which may stain the section | PAS reaction |
| for color development, wash in tap water after the sulfite rises | PAS reaction |
| Glutaraldehyde not recommended form fixation, it might react with schiff's reagent | PAS reaction |
| a control slide should be run through all steps of the procedure excep the periodate oxidation step | PAS reaction - to determine if there is any previously reactive aldehyde groups present in the tissue |
| chromate-containing fixatives may overoxidize rective groups during fixation the resulting reaction with schiff reagent may be weak | PAS reaction |
| Stains nerve fibers, nerve endings, neurofibrils | Bodian |
| demonstrate polysaccharides, neutral mucosubstances and basement membranes | PAS reaction |
| oxidation of certain tissue elements to aldehydes by periodic acid. | PAS reaction |
| reactive group is the 1,2 glycol group, other groups are also oxidized | PAS reaction |
| treat basic fuchsin w/sulfurous acid | preparing schiffs reagent |
| reduction casues loss of quinoid structures and masking of the chromophores | PAS reaction |
| colorless compound | leucofuchsin |
| following schiffs reagent, washing in running water causes loss of the bound sulfurous acid group attached at teh centra carbon atom, restoration of quinoid structure in the dye bound tby the aldehyde and the visualization of the typical shiff color | PAS reaction |
| Metabisulfate rinses are used to remove excess schiff reagent and prevent false colorization of the tissue elements due to oxidation of adsorbed reagent | PAS reaction |
| Fixation - 10% NBF or Bouin. blood smears should be fixed in methyl alcohol for 10 to 15 minutes | PAS reaction |
| 4 to 5 microns | PAS reaction |
| Control - section of kidney (most sensitive control). | PAS reaction |
| if demonstrating glycogen use liver containing glycogen or a section of cervix(include both endo and ecto cervix) | PAS reaction |
| Glycogen, neutral mucosubstances, certai epi sulfomucins and sialomucins, colloid material of thyroid, pars intermedia of pituitary, basement membranes, fungal walls | All show a positive PAS raction (Bright rose) |
| schiff staining after periodate does not demonstrate presence of carbohydrate residues | PAS reaction |
| abscence of staining doesnt mean tht carb residues are absent | PAS reaction |
| number of available 1,2 glycol groups, reactivity of schiff reagent w/reaction product, structure of polymer, exact procedural reaction conditions | intensitity of staining in routine PAS reaction is due to the folling factors |
| fast green counterstain m/b used instead of hematoxylin. when stain is used to demonstrate fungus | PAS reaction |
| sulfite rinse essential to remove any uncombined leucofuchsin following exposure to schiff's reagent | PAS reaction |
| highly chlorinated water is capable of oxidation. if section is transferred directly into tap water, any adsorbed schiff reagent m/b reoxidized to basic fuchsin,which may stain the section | PAS reaction |
| for color development, wash in tap water after the sulfite rises | PAS reaction |
| Glutaraldehyde not recommended form fixation, it might react with schiff's reagent | PAS reaction |
| a control slide should be run through all steps of the procedure excep the periodate oxidation step | PAS reaction - to determine if there is any previously reactive aldehyde groups present in the tissue |
| chromate-containing fixatives may overoxidize rective groups during fixation the resulting reaction with schiff reagent may be weak | PAS reaction |
| Liver containing large amounts of glycogen should not be used as control for | PAS reaction |
| demonstrate polysaccharides, neutral mucosubstances and basement membranes | PAS reaction |
| oxidation of certain tissue elements to aldehydes by periodic acid. | PAS reaction |
| reactive group is the 1,2 glycol group, other groups are also oxidized | PAS reaction |
| treat basic fuchsin w/sulfurous acid | preparing schiffs reagent |
| reduction casues loss of quinoid structures and masking of the chromophores | PAS reaction |
| spirochetes and bacteria - brown to black | dieterle |
| following schiffs reagent, washing in running water causes loss of the bound sulfurous acid group attached at teh centra carbon atom, restoration of quinoid structure in the dye bound tby the aldehyde and the visualization of the typical shiff color | PAS reaction |
| Metabisulfate rinses are used to remove excess schiff reagent and prevent false colorization of the tissue elements due to oxidation of adsorbed reagent | PAS reaction |
| Fixation - 10% NBF or Bouin. blood smears should be fixed in methyl alcohol for 10 to 15 minutes | PAS reaction |
| 4 to 5 microns | PAS reaction |
| Control - section of kidney (most sensitive control). | PAS reaction |
| if demonstrating glycogen use liver containing glycogen or a section of cervix(include both endo and ecto cervix) | PAS reaction |
| Glycogen, neutral mucosubstances, certai epi sulfomucins and sialomucins, colloid material of thyroid, pars intermedia of pituitary, basement membranes, fungal walls | All show a positive PAS raction (Bright rose) |
| schiff staining after periodate does not demonstrate presence of carbohydrate residues | PAS reaction |
| abscence of staining doesnt mean tht carb residues are absent | PAS reaction |
| number of available 1,2 glycol groups, reactivity of schiff reagent w/reaction product, structure of polymer, exact procedural reaction conditions | intensitity of staining in routine PAS reaction is due to the folling factors |
| fast green counterstain m/b used instead of hematoxylin. when stain is used to demonstrate fungus | PAS reaction |
| sulfite rinse essential to remove any uncombined leucofuchsin following exposure to schiff's reagent | PAS reaction |
| highly chlorinated water is capable of oxidation. if section is transferred directly into tap water, any adsorbed schiff reagent m/b reoxidized to basic fuchsin,which may stain the section | PAS reaction |
| for color development, wash in tap water after the sulfite rises | PAS reaction |
| Glutaraldehyde not recommended form fixation, it might react with schiff's reagent | PAS reaction |
| a control slide should be run through all steps of the procedure excep the periodate oxidation step | PAS reaction - to determine if there is any previously reactive aldehyde groups present in the tissue |
| chromate-containing fixatives may overoxidize rective groups during fixation the resulting reaction with schiff reagent may be weak | PAS reaction |
| Liver containing large amounts of glycogen should not be used as a control, the reaction can be weak but still very apparent | PAS reaction |
| reagent problems are apparent if a sectio n of kidney is used a s a control, when the reaction is intended for substances other than glycogen | PAS reaction |
| demosntrates glycogen in tissue | PAS W/diastase |
| very sensitive histochemical method for glycogen. diastase and amylase act on glycogen to depolymerize it into smaller units that are washed out of the section. | PAS W/diastase |
| Fixation - 10% NBF | PAS W/diastase |
| 4 to 5 microns | PAS W/diastase |
| control - section of liver containing glycogen, label with and without diastase | PAS W/diastase |
| contorl - cervix is also an excellent control (including both endo and ectocervix) | PAS W/diastase |
| Glycogen will stain brigh rose on section labeld without and will be absent from section labeled with | PAS W/diastase |
| glycogen fixed in picric acid containing fixatives may be more resistant to diastase digestion | PAS W/diastase |
| stratified squamous epi of the ectocervix (glycogen) and glands of the endocervix (mucin) will show a positive schiff reaction on slides without digestion, | PAS W/diastase |
| on slides with digestion, the stratified squamous epi will be negative while the gland of the endocervix remain positive | PAS W/diastase |
| used to differntiate between collagen nd smooth muscle in tumors and identify inreasees in collagenous tissue in disease such as cirrhosis of the liver | Masson trichrome stain |
| Thicrhome = three dyes | Masson trichrome stain |
| sections are stained w/ acid dyes, acidophillic elements like, cytoplasm, muscel, collagen wil bind w/acid dyes | Masson trichrome stain |
| sections are treated with phosphotungsic or phosphomolybdic acids | Masson trichrome stain |
| cytoplasm is less permealbe than collagen, phosphotungsic acid and phosphomolybdic acids cause biebrich scarlet to diffuse out the collagen but not the cytoplasm | Masson tricrhome stain |
| have numerous acid groups that most likely act as a link between the decolirzed collagen and aniline blue, the collagen dye | phosphotungsic and phosphomolybdic acids |
| pH of phosphotungsic/phosphomolybdic acids solution increases selectivity of collagen staining and aids in diffusion or removal of biebrich scarlet | Masson trichrome stain |
| Fixation - bouin is preferred, 10% NBF may be used | Masson trichrome stain |
| 4 to 5 microns | Masson trichrome stain |
| Control - every tissue has internal contorl, no other control sections are needed | Masson trichrome stain |
| control - if a control is desired, use uterus, small intestine, appendix or fallopian tube | Masson trichrome stain |
| Nuclei - black | Masson trichrome stain |
| Cytoplasm, keratin, muscle fibers - red | Masson trichrome stain |
| Collagen and mucus - blue | Masson trichrome stain |
| collagen may be stained with light green instead of aniline blue | Masson trichrome stain |
| light green is better counter stain when collagen is predominent, when small amounts are to be demonstrated aniline blue is better | Masson trichrome stain |
| decreased red staining indicates the staining solution has aged or overused, should be discarded | Masson trichrome stain |
| If blue staining of CT tissue appears faded, section has been overdifferentiated in acetic acid | Masson trichrome stain |
| altered collagen (burns) may lose affinity for aniline blue and bind to the acid dye instead | Masson trichrome stain |
| Picric acid w/less than 10% water is very explosive, its important that solution not be spilled in oven and allowed to evaporate | Masson trichrome stain |
| staining jar containing picric acid should be placed inside another container while in oven | Masson trichrome stain |
| iron hematoxylin is used for nuclear staining in trichrome procedure bcuz iron hematoxylin is more resistant than aluminum hematoxylin to decolorization b in acidic dye solution | Masson trichrome stain |
| Nuclei - dark blue | Masson trichrome stain |
| cytoplasm, keratin, muscle fibers - red | Masson trichrome stin |
| collagen and mucus - blue | Masson trichrome stain |
| indentify increase in collagenous CT fibers or differentiate between collagen and smooth muscle fibers | Gomori one step trichrome |
| plasma stain and CT fiber stain are combined in a soludtion of phosphotungsic acid to which glacial acetic acid has been added | Gomori one step trichrome |
| phosphotungsic acid favors the red staining of the muscle and cytoplasm. | Gomori one step trichrome |
| The tungstate ion is taken up by collagen and the CT fiber stain is subsequently bound to this complex, coloring the collagen green or blue, | Gomori one step trichrome |
| Fixation - any well fixed tissue may be used, Bouin solution is used as a mordant to intensify the color reactions. | Gomori one step trichrome |
| 4 to 5 microns | Gomori one step trichrome |
| Control - every tissue has internal contorl, no other control sections are needed | Gomori one step trichrome |
| control - if a control is desired, use uterus, small intestine, appendix or fallopian tube | Gomori one step trichrome |
| Nuclei - black | Gomori one step trichrome |
| cytoplasm, keratin, muscle fibers - red | Gomori one step trichrome |
| Collagen and mucus - green or blue | Gomori one step trichrome |
| used to demonstrate pathological changes in elastic tissue | Verhoeff elastic stain |
| demonstrates atrophy of tissue, thinnin/loss that results from arteriosclrotic changes and reduplicarion, breaks/splitting from vascular disease. | Verhoeff elastic stain |
| also used to demonstrate normal elastic tissue like identification of veins and arteries to determine if the blood vessels are invaded by tumors | Verhoeff elastic stain |
| tissue is overstained w/soluble lake hematoxylin-ferric chloride-iodine. | Verhoeff elastic stain |
| ferric chloride and iodine serve as mordants, they also have oxidizing function that assists in convervting hematoxylin to hematein | Verhoeff elastic stain |
| this method requires that sections be overstained and then differentiated it is REGRESSIVE | Verhoeff elastic stain |
| differentiation is accomplished by using excess mordant, ferrich chloride, to break the tissue-mordant-dye lake complex | Verhoeff elastic stain |
| breaking the tissue-mordant-dye lake complex allows other elements to be decolorized and the elastic fibers to remain stained | Verhoeff elastic stain |
| sodium thiosulfate is used to remove excess iodine. van Gieson solution is most commonly used counterstain. others m/b used | Verhoeff elastic stain |
| Fixation - any well-fixed tissue m/b used. neutral buffered formalin or zenker is preferred | Verhoeff elastic stain |
| 4 to 5 microns | Verhoeff elastic stain |
| control - aorta embedded on edge or cross section of large artery | Verhoeff elastic stain |
| elastic fibers - blue-black to black | Verhoeff elastic stain |
| Nuclei - blue to black | Verhoeff elastic stain |
| Collagen - red | Verhoeff elastic stain |
| other tissue elements - yellow | Verhoeff elastic stain |
| easy to overdifferntiate this stain | Verhoeff elastic stain |
| overdifferentiated sections m/b restained at any step, if the have not been treated w/alcohol | Verhoeff elastic stain |
| dont prolong staining w/van Gieson, as picric acid also will differentiate the stain further | Verhoeff elastic stain |
| not necessary to remove mercury deposits, they will be removed by staining solution | Verhoeff elastic stain |
| preparation of van Gieson solution is critical for proper differentiation of muscle and collagen | Verhoeff elastic stain |
| if picric acid is not saturated, collagen will not stain red and cytoplasm, muscle and collagen may all stain the same color | Verhoeff elastic stain |
| To prepare Verhoeff elastic stain solution, the reagents must be added in the order give w/mixing after each addition or poor stain w/result | Verhoeff elastic stain |
| for optimum results, slides must be individually differentiated, as time of differentiation is some what dependent on amount of elastic tissue present | Verhoeff elastic stain |
| dont depend on control for timing the differentiation of all sections | Verhoeff elastic stan |
| Bcuz proper differentiation is sometimes difficult, it is helpful to use duplicate sections differentiated to a slightly different end point | Verhoeff elastic stain |
| used to demonstrate pathological changes in elastic tissue | Aldehyde fuchsin elastic stain |
| also used to demonstrate normal elastic tissue like identification of veins and arteries to determine if the blood vessels are invaded by tumors | Aldehyde fuchsin elastic stain |
| Hydrochloric acid and paraldehyde are added to an alcoholic solutin of basic fuchsin to form aldehyde fuchsin | Aldehyde fuchsin elastic stain |
| schiff bases are formed by aldehyde and the fuchsin, but the affinity of elastic fibrs for this solution is not understood | Aldehyde fuchsin elastic stain |
| other tissue elements will also stain, includes pancreatic beta cell granules and sulfated mucosubstances. staining is intensified by prior oxidation | Aldehyde fuchsin elastic stain |
| Fixation - neutral buffered formalin preferred. Chormate fixatives should be avoided | Aldehyde fuchsin elastic stain |
| Formalin and bouin fixed tissues will show a colorless background. mercury fixed tissue will show a pale lilac background | Aldehyde fuchsin elastic stain |
| 4 to 5 microns | Aldehyde fuchsin elastic stain |
| Control - section of aorta embedded on edge or a cross section of large artery. skin also provides good control | Aldehyde fuchsin elastic stain |
| elastic fibers - deep blue to purple | Aldehyde fuchsin elastic stain |
| Other tissue elements - Green | Aldehyde fuchsin elastic stain |
| paraldehyde used for preparation of aldehyde fuchsin should be fresh | Aldehyde fuchsin elastic stain |
| old solution of aldehyde fuchsin doesnot stain well, staining time may need to be prolonged | Aldehyde fuchsin elastic stain |
| do not use rosanili, it is nt satisfactory | Aldehyde fuchsin elastic stain |
| shelf life aldehyde fuchsin may be prolonged by refrigerating a small amount and freezing aliquots of the remainder | Aldehyde fuchsin elastic stain |
| acetaldehyde may be used in place of paraldehyde, acetaldehyde is chaper and may be obtained without a DEA number | Aldehyde fuchsin elastic stain |
| demonstrate reticular fibers in tissue, important in differential dx of certain types of tumors | Gomori stain for reticular fibers |
| a change from normal reticular fiber pattern, seen in some liver disease | Gomori stain for reticular fibers |
| hexose sugars of reticulin are demonstrated by oxidation | Gomori stain for reticular fibers |
| potassium permangenate is oxidizing agent in this procedure and excess is removed by potassium metabisulfate | Gomori stain for reticular fibers |
| Ferric ammomium sulfate acts as the sensitizer and is replaced by silver from the diamine silver solution | Gomori stain for reticular fibers |
| following impregnation formalin is used to reduce the silver to its visible metallic form | Gomori stain for reticular fibers |
| toning with gold chloride and removal of unreacted silver with soidum thiosulfate. the final step is a counterstain if desired | Gomori stain for reticular fibers |
| Fixation - 10% NBF | Gomori stain for reticular fibers |
| 4 to 5 microns | Gomori stain for reticular fibers |
| Control - liver | Gomori stain for reticular fibers |
| Reticulin - black | Gomori stain for reticular fibers |
| Collagen - Taupe | Gomori stain for reticular fibers |
| a hint of turbidity should remain in the silver solution | Gomori stain for reticular fibers |
| excess of ammonia decrases the sensitivity and results in incomplete impregnation of reticular fibers | Gomori stain for reticular fibers |
| for good reticulin demonstration, rinse between the diamine silver solution and the formaldehyde | Gomori stain for reticular fibers |
| if wash is prolonged, staining of the reticulin will be reduced and if its insufficient there will be excessive background staining | Gomori stain for reticular fibers |
| glassware m/b chemically cleaned w/comercial cleaning agents or bleach | Gomori stain for reticular fibers |
| the older cleaining method of using a mix of sulfuric acid and potassium dichromate is no longer used bcuz of hazards | Gomori stain for reticular fibers |
| use plastic forceps in this method | Gomori stain for reticular fibers |
| aka iron alum. | ferric ammonium slufate |
| term used for salts that are double or bisulfates, like potassium aluminum sulfate, ammonium aluminum sulfate, chromium potassuim sulfate, and ferric ammonium sulfate | Alum |
| if nuclear fast red is the counterstain, wash the sildes well w/water after staining | Gomori stain for reticular fibers |
| if slides are not transfered from counterstain directly to alcohol, they will develop a cloudiness, that can be removed by backing the slide up to water | Gomori stain for reticular fibers |
| pattern of reticular fiber staining is very important, should be seen easy w/scanning lens of microscope (liver bx) | Gomori stain for reticular fibers |
| a guide to the quality of the stain is the easy visualization of reticulin pattern w/x 10 objective | Gomori stain for reticular fibers |
| counterstain, may obscure the easy visualization and therefore should not ve used when visualization of this pattern is important | Gomori stain for reticular fibers |
| If silver stain of nuclei is a problem, fix by using acetified potassium permanganate | Gomori stain for reticular fibers |
| if ammonium hydroxide looses strength, due to loss of ammonia from solution, open a fresh bottle | Gomori stain for reticular fibers |
| decrease in ammonia is noticed bcz more than the usual ammount of ammonium hydroxide is required | Gomori stain for reticular fibers |
| Chukurian uses ammonical silver solution for several days,storing it in fridge and brining it to room temp before use | Gomori stain for reticular fibers |
| Chukurian feels reticulin stains better after ammonical silver has aged for a few days | Gomori stain for reticular fibers |
| ammonical silver may form explosive compound, use care in preparation, use and storage of this solution | Gomori stain for reticular fibers |
| storage in regrigerator retards formation of explosive compound, avoid exposure to direct sunlight | ammonical silver solution |
| deposit of silver should be in a linear pattern | Gomori stain for reticular fibers |
| Demonstrate muscle cross striations and fibrin | Mallory PTAH |
| are a dx feature of rhabdomyosarcomas or tumors arising form striated muscle. | cross-striations |
| Nemaline present in some skeletal muscle dx, may also be demonstrated by this method | Mallory PTAH |
| amount of phosphotunstic acid in the staining solution is greater than the amount of hematein | Mallory PTAH |
| tungsten binds all available hematein to give a blue-colored lake | Mallory PTAH |
| metal hematein lake stains selected tissue components blue, while the phosphotungstic acid is thought to stain the red-brown components | Mallory PTAH |
| This stain is referred to as a polychrome stain bcuz one solutin gives two major colors | Mallory PTAH |
| components colored red-brown will lose this color with wateror prolonged alcohol washes, and therefore dehydration of section following staining must be rapid | Mallory PTAH |
| Fixation - Zenker preferred, 10% NBF m/b used | Mallory PTAH |
| 4 to 6 microns | Mallory PTAH |
| control - skeletal or cardiac muscle to demonstrate cross striations, | Mallory PTAH |
| control - section containing fibrin for demonstration of fibrin, or secion of cerebral cortex (not spinal cord) for demonstration of glial fibers | Mallory PTAH |
| Cross-striations , fibrins - blue | Mallory PTAH |
| Nuclei - Blue | Mallory PTAH |
| Collagen - red/brown | Mallory PTAH |
| PTAH has been replaced by IHC, although fibrin will stain well after formaldehyde fixation | Mallory PTAH |
| you get better results if tissue is fixed in Zenker on in a solution containing mercury | Mallory PTAH |
| stain is not as good when formalin fixed sections are mordanted as wehn original fixative is mercuric | Mallory PTAH |
| uneven staining occurs in formalin fixed tissue | Mallory PTAH |
| chemically oxidized staining solution has a shorter shelf life than naturally ripened staining solution bcz chemical oxidation may cause overoxidation. | Mallory PTAH |
| when this occurs there is failure to show proper density of th blue tones. | chemical oxidation may cause over oxidation. |
| solutions should be stored in amber glass bottles to retard overoxidation by light | Mallory PTAH |
| thorough washing of section before staining is essential, as hydration of tissue structures will greatly facilitate uptake of dye molecules | Mallory PTAH |
| sodium thiosulfate interferes w/binding of the PTAH, sections should be washe very well after application of sodium thiosulfate | Mallory PTAH |
| delineates glomerular basement membranes | Periodic acid-methenamine silver microwave (PAMS) |
| Methenamine silver demonstrates carbohydrate component of basement membrane by oxidizing the carbs to aldehydes | Periodic acid-methenamine silver microwave (PAMS) |
| in silver tecnique for staining basement membrane, silver ions from the methenamine silver complex are first bound to carb compnents of basement membrane and then reduced to visible metallic silver by aldehyde group | Periodic acid-methenamine silver microwave (PAMS) |
| toning is done with gold chloride and any unreduced silver is removed by sodium thiosulfate | Periodic acid-methenamine silver microwave (PAMS) |
| Fixation - 10% NBF preferred, mercury containing fixatives not recommended | Periodic acid-methenamine silver microwave (PAMS) |
| paraffin processed tissue cut at 2 microns | Periodic acid-methenamine silver microwave (PAMS) |
| Control - kidney has an internal control. no other control slide is necessary | Periodic acid-methenamine silver microwave (PAMS) |
| Basement membrane - black | Periodic acid-methenamine silver microwave (PAMS) |
| Background - green | Periodic acid-methenamine silver microwave (PAMS) |
| sharper staining of basement membrane and less background staining can be obtained with the use of microwave oven for silver stain | Periodic acid-methenamine silver microwave (PAMS) |
| temp is critical and should be below boiling, 95C, immediately after removal from oven. | Periodic acid-methenamine silver microwave (PAMS) |
| each oven should be calibrated for the time required to reach the correct temperature | Periodic acid-methenamine silver microwave (PAMS) |
| very difficult stain to perform correctly | Periodic acid-methenamine silver microwave (PAMS) |
| glomerular basement membrane should appear as continuous black line. stopping silver impregnation too soon results in uneven or interupted stain | Periodic acid-methenamine silver microwave (PAMS) |
| application of too much counterstain will mask the silver stain and decrease contrast | Periodic acid-methenamine silver microwave (PAMS) |
| demonstrate neutral lipids in frozen sections. | Oil red O |
| demonstrates fat occuring in abnormal places, such as fatty emboli that develop after bone fracture or injury that crushes fatty body area | Oil red O |
| fat verifies that emboli caused death | Oil red O |
| degenarating material containing fat, like cell membranes or myelin, may gro into fat droplets that are demonstrable with fat stain | Oil red O |
| tumors arising from fat cells (liposarcomas) can be differentiated from others with | Oil red O |
| staining with oil-soluble dye is based on greater solubility of the dye in the lipoid substances than in the usual hydroalcoholic dye solvent | Oil red O |
| this is a physical method of staining | Oil red O |
| dyes used must be more soluble in tissue lipid than in the solvent, must not be water soluble, m/b strongly colored, act w/tissue constituents by solution | Oil red O |
| solvent used is critical, w/isopropanol removing a minimal amount of lipid and propylene glycol not extracting any lipid | Oil red O |
| Fixation 10% NBF or calcium-formal, alcoholic fixatives should not be used, bcz of their lipid dissolving ability | Oil red O |
| cut frozen sections at 10 microns | Oil red O |
| Paraffin sections cant be used bcuz dehydrating and clearing agents dissolve the fat | Oil red O |
| if free-floating sections arnot used, sections of fixed tissue should be picked up on coated or subbed slides | Oil red O |
| Control- most tissue contain fat, a control isnot used | Oil red O |
| Fat - intense red | Oil red O |
| Nuclei - Blue | Oil red O |
| to improve microtomy of FZ, formalin fixed tissue may be infiltrated with 30% sucrose before freezing | Oil red O |
| aqueous mounting media must be used, organic solvent present in synthetic resinous media wll disolve fat | Oil red O |
| fat in section is relatively liquid and mobile, care should be taken so that no pressure is placed on the coverglss or the fat will be displaced | Oil red O |
| if air bubbles are present in section, remove cover slip by soaking the slide in warm water | Oil red O |
| If glycerine jelly is used for mountin, it should not be overheated, this may melt the fat and displace it | Oil red O |
| stain should be done at 60 degrees celsius | Oil red O |
| Demonstrates fungus | Grocott methanamine silver |
| Control section w/fungs, Pneunmocytis use pneumocytis control | Grocott methanamine silver |
| Principle - Polysaccharides in fungal cell wall oxidize to aldehyde by chromic acid | Grocott methanamine silver |
| Mucin - taupe to gray | Grocott methanamine silver |
| Background - green | Grocott methanamine silver |
| Fixation - 10% NBF | Grocott methanamine silver |
| 4 to 5 microns | Grocott methanamine silver |
| chemically cleaned glassware and nonmetallic forceps must be used | Grocott methanamine silver |
| Demonstrates spirochetes, hpylori, legionela and cat scratch fever | Steiner and Steiner |
| Principle - argyrophilic method, organisms have ability to adsorb silver ions but not reduce the silver to a visible form, a chemical reducer hydroquinone is used for that purpose | Steiner and Steiner |
| Legionella, hpylori, spirochetes, pneumophila, and other bacteria - dark brown to black | Steiner and Steiner |
| Background - light yellow | Steiner and Steiner |
| demonstrate spirochetes | Warthin starry |
| Principle - argyrophilic method, spirochetes have ability to bind silver ions but not reduce the silver to a visible form, a chemical reducer hydroquinone is used for this purpose | Warthin starry |
| spirochetes - black | Warthin starry |
| Other bacteria - black | Warthin starry |
| Background - Pale yellow to light brown | Warthin starry |
| Control - section with spirochetes | Warthin starry |
| indication of reducing substances present in tissue. Melanin, argentaffin granules and formalin pigments w/b stained | Schmorl |
| reduces substances present in tissue. reduce the ferric ions present in staining solution to ferrous ions. | Schmorl |
| The ferrous ions combine w/t ferricyanide present in the staining solution to form a precipitate of ferrous ferricyanide (turnbull blue | Schmorl |
| Control - seciont w/ melanin or agentaffin granules | Schmorl |
| Reagents: Ferric Chloride stock solution, Potassium Ferricyanied stock solution, Ferric chloride-potassium ferricyaide working solution, mayer mucicarmine, metanil yellow | Schmorl |
| silver proteinate is usted to impregnate tissue sections, copper is added to the impregnating solution to "destain" connective tissue allows differtiation neural vx CT elements | Bodian |
| 6 to 8 | Bodian |
| demonstrate nerve fibers and nurofibrils | Holmes silver nitrate method |
| argyrophil method, requires chemical reduction.Gold chloride, oxalic acid,sodium thiosulfate | Holmes silver nitrate method |
| is more soluble in the lipids of cell wall than acid alcohol, but is readily removed from bacterias that lack the waxy capsule | Carbol fuchsin (kinyoun) |
| staining is enhanced by phenol and alchol, both of these chemicals help to disolve the basic fuchsin | Kinyoun acid fast stain |
| identify mycobacteria | Carbol fuchsin (kinyoun) |
| makes acid fast organisms non-acid | carnoy |
| lipoid capsule of mycobacteria is of high molecular weight. It is waxy at room temp, successful penetration by aqueous based staining solutions in gram staining is prevented | Kinyoun acid fast stain |
| overcounterstaining w/methylene blue will mask any organisms | Kinyoun acid fast stain |
| wash out acid before counterstaining or tissue will not stain | Kinyoun acid fast stain |
| This method is not satisfactory for mycobacterium leprea | Kinyoun acid fast stain |
| if the section is allowed to dry after carbol fuchsin application, a compound resistant to decolorization w/b formed | Kinyoun acid fast stain |
| detect presence of acid-fast mycobacteria | Ziehl-neelsen method for acid fast bacteria |
| lipoid capsule of acid-fast organism takes up carbol-fuchsin and resists decolorization w/dilute mineral acid | Ziehl-neelsen method for acid fast bacteria |
| acid fast bacteria - bright red, background - light blue | Ziehl-neelsen method for acid fast bacteria |
| This carbol-fuchsin method is preferred | Ziehl-neelsen method for acid fast bacteria |
| detect the presence of mycobacterium leprea (leprosy) | Fite acid-fast stain |
| lipoid capsule of organism takes up carbol fuchsin and resists decolorization with dilue mineral acid | Fite acid-fast stain |
| norcadia will also stain with | Fite acid-fast stain |
| fixation any except carnoy | Fite acid-fast stain |
| acid fastness of leprosy organism is enhanced when the waxy capsule is protected by the mixutre of peanut oil and xylene. avoid dehydration solutions | Fite acid-fast stain |
| detect mycobacteria tuberculosis or other acid fast organisms | Auramine-Rhodamine fluorescence |
| the exact mechanism of the stain is unknown. both of the dyes use basic dyes that fluoresce at short wave lengths. when both dyes are used toghether, they yield a better stain | Auramine-Rhodamine fluorescence |
| is more soluble in the lipids of cell wall than acid alcohol, but is readily removed from bacterias that lack the waxy capsule | Carbol fuchsin (kinyoun) |
| staining is enhanced by phenol and alchol, both of these chemicals help to disolve the basic fuchsin | Kinyoun acid fast stain |
| identify mycobacteria | Carbol fuchsin (kinyoun) |
| makes acid fast organisms non-acid | carnoy |
| lipoid capsule of mycobacteria is of high molecular weight. It is waxy at room temp, successful penetration by aqueous based staining solutions in gram staining is prevented | Kinyoun acid fast stain |
| overcounterstaining w/methylene blue will mask any organisms | Kinyoun acid fast stain |
| wash out acid before counterstaining or tissue will not stain | Kinyoun acid fast stain |
| This method is not satisfactory for mycobacterium leprea | Kinyoun acid fast stain |
| if the section is allowed to dry after carbol fuchsin application, a compound resistant to decolorization w/b formed | Kinyoun acid fast stain |
| detect presence of acid-fast mycobacteria | Ziehl-neelsen method for acid fast bacteria |
| lipoid capsule of acid-fast organism takes up carbol-fuchsin and resists decolorization w/dilute mineral acid | Ziehl-neelsen method for acid fast bacteria |
| acid fast bacteria - bright red, background - light blue | Ziehl-neelsen method for acid fast bacteria |
| This carbol-fuchsin method is preferred | Ziehl-neelsen method for acid fast bacteria |
| detect the presence of mycobacterium leprea (leprosy) | Fite acid-fast stain |
| lipoid capsule of organism takes up carbol fuchsin and resists decolorization with dilue mineral acid | Fite acid-fast stain |
| norcadia will also stain with | Fite acid-fast stain |
| fixation any except carnoy | Fite acid-fast stain |
| acid fastness of leprosy organism is enhanced when the waxy capsule is protected by the mixutre of peanut oil and xylene. avoid dehydration solutions | Fite acid-fast stain |
| detect mycobacteria tuberculosis or other acid fast organisms | Auramine-Rhodamine fluorescence |
| the exact mechanism of the stain is unknown. both of the dyes use basic dyes that fluoresce at short wave lengths. when both dyes are used toghether, they yield a better stain (auramine O, rhodamine B) | Auramine-Rhodamine fluorescence |
| false-positive may occure | Auramine-Rhodamine fluorescence |
| slides stained with auramine O-rhodamine c/b restained with carbol-fuchsin for confirmation if the results are questionable | Auramine-Rhodamine fluorescence |
| this method is more likely to stain dead or dying organisms than the carbol-fuchsin | Auramine-Rhodamine fluorescence |
| use small amounts of Rhodamine B it will intesify the fluorescence of mycobacteria. too much Rhodamine B will quench fluoresence, even in low concentration | Auramine-Rhodamine fluorescence |
| fluorescence micropscopy is not satisfactory after fixation in solutions w/heavy metals. heavy metals tend to quench the primary fluorescence of the specimen | Auramine-Rhodamine fluorescence |
| Modification of Gram | Brown-Hopps |
| to demonstrate Gram-negative and Gram-positive bacteria in tissue | Brown-Hopps |
| crystal violet is applied first followed by an iodine mordant forming a dye lake. | Brown-Hopps |
| both have a cell wall composed of peptidoglycan | Gram-negative and Gram-positive bacteria (Brown-Hopps) |
| the wall of gram-positive bacteria is thicker than the wall of gram-negative organisms | Gram-negative and Gram-positive bacteria (Brown-Hopps) |
| contains a layer of lipopolysaccharide external to cell wall | gram-negative bacteria (Brown-Hopps) |
| cannot be washed out of the peptidoglycan layer of gram positive cells | large crystal violet-iodine comlex |
| can be removed from gram-negative bacteria, bcuz alcohol and acetone disrupt the outer lipopolysaccharide layer | large crystal violet-iodine comlex |
| this method is preferred for staining gram-negative organisms and ricketssiae | Brown-Hopps |
| this is a useful method for for screening of infectious agents that cause actinomycosis, nocardiosis, coccidiomucosis, blastomycosis, aspergillosis, rhinosporidiosis and amebiasis | Brown-Hopps |
| gram-positive bacteria -blue, gram-negative bacteria -red, background -yellow, nuclei -light red | Brown-Hopps |
| can be used for the demonstration of bacteria, ricketssias and toxoplasma gondii. | Giemsa |
| identification of helicobacter pylory | Modified diff quick Giemsa |
| the Romanowsky stains "neutral" dyes combining the basic dye methylene blue and the acid dye eosin. methylene blue and eosin give a wide color range when staining tissues and blood smears. | Modified diff quick Giemsa |
| hpylori - dark blue, other bacteria -blue, nuclei -dark blue, cytoplasm -pink | Modified diff quick Giemsa |
| demonstrate fungi in tissue | Hotchkiss-Mcmanus Pas reaction |
| "carbohydrates". polysaccharides present in fungal cell walls are oxidized by periodic acid to aldehyes. the aldehydes react w/schiffs reagent to yield a rose colored fungi | Hotchkiss-Mcmanus Pas reaction |
| fungi -rose, background -green | Hotchkiss-Mcmanus Pas reaction |
| when staining for fungus, the green counterstain provides better contrast | Hotchkiss-Mcmanus Pas reaction |
| it is helpful to use diastase digestion in sections containing glycogen | Hotchkiss-Mcmanus Pas reaction |
| oxidizing and schiffs reagent must not be overused or you will get a poor stain | Hotchkiss-Mcmanus Pas reaction |
| demonstrates fungus in tissue | Gridley |
| this is a modification of Bauer | Gridley |
| use chromic acid to oxidize adjacent glycol groups to aldehydes. the aldehydes react w/schiffs reagent. aldehyde fuchsin acts as an aldehyde and occupies uninvolved linkages to schiffs. | Gridley |
| is a stronger oxidizing agent than periodic acid. it further attacks and destroys aldehydes | chromic acid |
| very old nonviable fungus doesnot stain well with this technique as with GMS | Gridley |
| sulforous acid rinse m/b used after schiffs | Gridley |
| coccidioides immitis | Gridley |
| melanin granules, chromatin, formalin pigments and foreign material present in macrophage will stain brown to black, | dieterle |
| QC- tissue containing, hpylori, legionella, pneumophila | Steiner and Steiner |
| chromic acid is a strong oxidant. it oxidizes many newly released aldehyde groups. it breaks down products that will not react. this helps to suppress the weak background reactions of collagen fibers and basement membranes | chromic acid (GMS) |
| fungal cell walls, glycogen and mucins are substances that hvae large quantities of polysaccharide. they remain reactive with methanamine silver reducing it to visible metallic silver | Grocott Methanamine-silver nitrate |
| Methanamine gives the solution the alkaline properties necessary for proper reaction, and sodium borate acts as the buffer. gold chloride tones, and sodium thiosulfate removes unreduced silver | Grocott Methanamine-silver nitrate |
| QC- if staining for pneumocystis. use pneumocystis control | Grocott Methanamine-silver nitrate |
| background -pale yellow, or tan | dieterle |
| Methanamine silver decomposess at higher temperatures | Grocott Methanamine-silver nitrate |
| Gluteraldehyde should be avoided. Bcuz teh free aldehyde groups reduce the silver and give w/g you nonspecific staining | Grocott Methanamine-silver nitrate |
| failing to remove alc used in deparaffination and hydration w/result in reduction of chromic acid. this causes the color to change from orange to brown. | Grocott Methanamine-silver nitrate |
| it requires takes longer for p carnii to stain. if unsure of what you are staining for, use a p carni control | Grocott Methanamine-silver nitrate |
| a water bath instead of oven should be used | Grocott Methanamine-silver nitrate |
| do not overheat silver or breaking down of solution will occur and staining m/b nonspecific | Grocott Methanamine-silver nitrate |
| may be used to demonstrate fungi, p Carinii, Actinomyces and related species, Nocardia asteroides and certain encapsulated bacteria | Grocott Methanamine-silver nitrate |
| bcuz of toxicity of chromic acid, periodic acid m/b used however, false negatives m/b obtained. | Grocott Methanamine-silver nitrate |
| sections that have been stained using other methods m/b used with GMS. Any exisiting stain w/b removed by chromic acid | Grocott Methanamine-silver nitrate |
| Rapid method, useful on cytospin preparations or on F.S. for dx of Pneumocystis carinii. is also good as a more routine procedure | Microwave Methanamine-silver nitrate procedure |
| fixation: cytospin -95%alc, F.S. -40%formaldehyde, paraffin sections fxd 10%NBF | Microwave Methanamine-silver nitrate procedure |
| F.s 6um, parrafin 4 to 5Um | Microwave Methanamine-silver nitrate procedure |
| place plastic coplin jars in loosely closed plastic bag in the microwave oven. any solution that boils over will be left in the bag | Microwave Methanamine-silver nitrate procedure |
| reticular fibers, rbc's, and other tissue w/stain if the slides are left in methanamine-silver to long. this nonspecific staining will mimic the appearance of fungi and will also obscure any fungi that is present | Microwave Methanamine-silver nitrate procedure |
| chromic acid is very toxic. instead of microwaving, oxidation c/b done at 58C for 10 minutes. | Microwave Methanamine-silver nitrate procedure |
| demonstrate the capsule of cryptococcus | Mayers mucicarmine and alcian blue |
| the mucoid capsule of cryptococcus neoforma c/b differentiated form other yeast like fungi of similar size and shape | Mayers mucicarmine and alcian blue |
| if sections over developed, treat with iodine and sodium thiosulfate for color removal | Warthin starry |
| all bacteria are nonselectively blackened by silver impregnation methods, | Warthin starry, Dieterle, and Steiner and Steiner |
| demonstrate small, wealky gram negative bacterial such as legionella. | Warthin starry, Dieterle, and Steiner and Steiner |
| provide much greater sensitivity twhen screening for small numbers of gram pos and gram neg bacteria | silver impregnation |
| demonstrate spirochete and other bacteria in tissue | microwave modification of warthin starry |
| alipia felis (cat scratch bacillus)-black, legionella pneumophila -black, nocardia asteroides -black, hpylori -black, nuclei brown, erythrocytes -brown | microwave modification of warthin starry |
| results are more consistent and reliable when staining and developing are done at lowest power levels | microwave modification of warthin starry |
| spirochetes are not well demonstrated by this method | microwave modification of warthin starry |
| demonstrates spirochetes or the causitive organisms of legionellosis | dieterle |
| spirochetes are argyrophilic, the will adsorb silver from a silver solution but the adsorbed silver must be chemically reduced to visible metallic form. Hydroquinone is the reducing agent/developer | dieterle |
| QC-tissue containing spirochetes or legionella | dieterle |
| considered a primary CT stain, but rarely used. Its an excellent counterstain for other methods such as Verhoeff elastic | Van Gieson Picric Acid-Acid Fuchsin Stain |
| in strongly acidic solution, collagen is stained by acid fuchsin. picric acid provides pH and acts a s the stain for muscle and cytoplasm. addding hydrochloric acid helps to sharpen the differntiation btwn collagen and muscle | Van Gieson Picric Acid-Acid Fuchsin Stain |
| demonstrate mucin, fibrin, elastic fibers, muscle and collagen | Russell modification of the movat pentachrome stain |
| acidic mucosubstances are stained by AB. the alkaline alcohol solution coverts the AB to Moastral fast blue, which is insoluble.Iron hematoxylin is used to stain elastic fibers which are then differentiated w/ferric chloride, | Russell modification of the movat pentachrome stain |
| uses potassium ferrOcyanide | Prussian blue |
| Uses potassium ferrIcyanide | Turnbull blue |
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