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89 Cards in this Set
- Front
- Back
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Glycogen is best demonstrated by the use of:
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PAS with and without diastase
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Amyloid can be demonstrated with:
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Congo red
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The Schiff reaction demonstrates:
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aldehydes
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A good control for the Mayer mucicarmine stain is:
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appendix
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A good control for glycogen is:
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liver
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To increase the specificity for amyloid, Congo red stains should be examined by which microscope?
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polarizing
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Of the following, which is the best fixative for glycogen?
a) Zenker b) Orth c) acetic acid d) absolute alcohol |
d) absolute alcohol
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Periodic acid is used in the PAS technique as a/an:
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oxidizer
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The crystal violet stain for amyloid is a/an:
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polychromatic stain
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Acid mucopolysaccharides are demonstrated by:
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alcian blue
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The reliability of the Schiff reagent may be checked by adding which of the following to a small aliquot of the schiff solution?
a) sodium iodate b) formaldehyde c) potassium metabisulfite d) diastase |
b) formaldehyde
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A fluorescent dye used for the demonstration of amyloid is:
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thioflavin T
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The alcian blue stain performed at pH 1.0 demonstrates:
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sulfated acid mucopolysaccharidess
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Substances stained positively with the colloidal iron will be:
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blue
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Schiff reagent is a/an:
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reduced solution of basic fuchsin
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Diastase digestion increases specificity for:
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glycogen
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Colloidal iron is used for the demonstration of:
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acid mucopolysaccharides
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A mucicarmine stain would be most helpful in the diagnosis of:
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adenocarcinoma
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Adjacent sections are stained with PAS, one with and one without diastases digestion. A positive result on the one without digestion and a negative result on the one with digestion indicates the presence of:
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glycogen
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T/F: Glucose, sucrose, and other oligopolysaccharides can be demonstrated easily in tissue sections.
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false
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T/F: Hyaluronidase is used to digest some connective tissue mucins.
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true
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T/F: The routine alcian blue stain is done at pH 1.5.
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false
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T/F: Good Schiff reagent should be light pink.
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false
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T/F: The end product in the colloidal iron method is Prussian blue.
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true
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T/F: Amyloid shows a yellow birefringence following staining with Congo red.
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false
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T/F: Glutaraldehyde is one of the recommended fixatives for the PAS reaction.
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false
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T/F: The Schiff reaction may show false positively following chromate-containing fixatives.
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false
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T/F: Glycogen-containing tissue fixed in Bouin solution may show resistance to diastase digestion.
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true
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T/F: The addition of acid to the crystal violet staining solution reduces background staining.
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true
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T/F: 4um to 6um sections are recommended for crystal violet stains.
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false
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What reagents are used to show acid mucopolysaccharides?
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-alcian blue, pH 1.0
-alcian blue, pH 2.5 -colloidal iron |
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What reagents are used to show amyloid?
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-Congo red
-crystal violet -thioflavin T |
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What reagents are used to show carboxylated acid mucopolysaccharides?
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-alcian blue, pH 2.5
-colloidal iron |
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What reagents are used to show glycogen?
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-diastase
-periodic acid -Schiff reagent |
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What reagents are used to show neutral polysaccharides?
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-periodic acid
-Schiff reagent |
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Which reagents are used to show sulfated (only) acid mucopolysaccharides?
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-alcian blue, pH 1.0
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No staining of the glomerular basement membrane can be seen microscopically on a control section of kidney. This may be the result of:
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inadequate oxidation
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Marked nonspecific background staining is noted on a section stained with the PAS technique. This could be the result of:
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fixation with glutaraldehyde
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Very weak staining is noted on a PAS-stained control section of liver. One problem-solving action is to:
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check the Schiff reagent with formaladehyde
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Sections of small intestine show orange goblet cells that are partially obscured by the yellow background. This is most likely the result of:
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overstaining with metanil yellow
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Control sections stained with Congo red show only yellow, and no green, birefringence. This could probably be corrected in the future by:
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ensuring that the sections are cut at 8um to 10um
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what are examples of neutral epithelial mucins
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ovimucoid
stomach mucin Paneth call granules |
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What are examples of COOH and OSO3H acid connective tissue mucins
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Chondrotin sulfate A, B, C
Heparin |
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Reagent order in PAS
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P-periodic
A-acid S-schiff metabisulfite rinse Harris hematoxylin |
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What is the Schiff reagent made of
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basic fuchsin (pararosaniline)
charcol HCl metabisulfite |
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Factors affecting PAS stain
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amount of 1,2 glycols avaliable
reactivity of Schiff reagent |
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The PAS is what type of reaction
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oxidation
1,2 glycol to aldehyde |
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Fixative for PAS
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10% NBF
Bouins methanol- blood |
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Which fixative is not good for the removal of glycogen in the PAS with digestion stain?
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picric acid fixative will be most resistant to glycogen removal
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Facts about mucins
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mucins: stain with basic dyes
metachromatic precipitated by acetic acid soluble in alkaline solutions |
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What forms a compound in the Mayer Muccicarmine stain?
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Aluminum forms a chelation with carmine; compound has a + charge attaches to acid groups of mucin.
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Which stain detects epithelial mucins?
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Mayer Muccicarmine
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What is muccicarmin composed of?
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carmine
aluminum hydroxide alcohols |
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Steps of Mayer Muccicarmine stain
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Weigerts Iron Hematoxylin
Muccicarmine Menatil Yellow |
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Which stains detect glycogen?
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PAS
Best Carmine |
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What is important in the Best Carmine reaction?
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high pH basic
ammonium maintains high pH |
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Procedure for Best Carmine
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Harris Hematoxylin
Carmine Alcohol |
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What is the differentiating solution in Best Carmine?
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alcohol
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Types of connective tissue mucin stains
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Alcian Blue
Colloidal Iron |
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If there is weak stainig in Alcain Blue, what may cause this?
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complete hydration of slide necessary b/c alcaniphilic structures hydrate slowly
weak staining will result if not properly hydrated |
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steps of Alcian Blue pH2.5
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Acetic Acid
Alcian Blue w/ acetic acid Acetic acid nuclear fast red |
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What does the acetic acid in Alcian Blue do?
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Acetic acid prepares the tissue to stain, protects pH changes, and prevents nonspecific staining
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steps of Alcian Blue pH1.0
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HCl
Alcian Blue w/ HCl HCl nuclear fast red |
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Which connective tissue stain can will not work with a chromate fixative?
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Collodial iron
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What reactions occurs in Colloidal Iron stain?
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Colloidal ferric iron at low pH binds;
Prussian blue detects the iron bound to the tissue |
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Steps of colloidal iron
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acetic acid
colloidal iron acetic acid Potassium ferrocyanide hydrochloric nuclear fast red |
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Whaich stain differentiate between connective tissue mucins and epithelial mucins
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Alcian Blue w digestion
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Alcian Blue/ PAS stain demonstrates
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differentiate between connective tissue (acid) mucins and neutral mucins
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What helps to enhance amyloid staining?
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alkali pretreatment makes more site avaliable for dye binding
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How to examine Congo Red results
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see results under polarized light
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Results of Congo Red stain
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Green birefringence
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Cut size for amyloid Congo Red tissue
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6-10um, if thinner will not show green birefringence
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Steps for Cyrstal Violet stain
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Crystal violet
Apathy Mounting Media |
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What prevents bleeding of the cyrstal violet stain
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potassium acetate or sugar
(mounting apathy medium contains sugars) |
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How is the background diminished in Thioflavin T stain
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Background nuclear staining is quenched by staining with aluminum hematoxylin (Mayer Hematoxylin)
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Which of the following will bind to acid mucosubstances and may then be demonstrated by the Prussian blue reaction?
a) diastase b) dimedone c) colloidal iron d) hyaluronidase |
c) colloidal iron
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Parallel sections are stained with PAS, one with and one without diastase digestion. When the staining results are evaluated, the digested section demonstrates:
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sites where glycogen was removed
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Acid mucosubstances and neutral mucosubstances can be differentiated by stained with both
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Acid mucosubstances and neutral mucosubstances can be differentiated by stained with both alcian blue and PAS
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In the colloidal iron method of staining, the principle of the reaction is believed to be the formation of an ionic bond between ferric iron and the free carboxyl group of the:
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acid mucopolysaccharides
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In order to suppress background and non-specific staining, a Congo red solution frequently contains:
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sodium chloride
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In metachromatic staining, the strong acid mucosubstances can be distinguished from weakly-acid mucosubstances by varying the:
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pH
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Carmine and aluminum chloride combine to form Mayer:
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mucicarmine
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The PAS reaction is useful for the demonstration of:
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neutral mucopolysaccharides
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Microscopic inspection of a PAS-stained control section for fungi reveals very palely stained fungal organisms, making identification difficult. A likely cause for this problem is the:
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use of depleated Schiff reagent
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The acid used in the Prussian blue reaction is:
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hydrochloric
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Tissue from which of the following organs should be selected as a control for the alcian blue technique (pH 2.5)?
a) skin b) kidney c) muscle d) colon |
d) colon
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An alcian blue stain at pH 2.5 has been requested, but no alcian blue is available. Which of the following procedures could be performed to give equivalent results?
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a) Luxol fast blue
b) periodic acid-Schiff c) colloidal iron d) mucicarmine |
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The major disadvantage of using crystal violet technique for the demonstration of amyloid is that:
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the preparation is not permanent
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What is potassium ferranocyde hydrochloric used for
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Colloidal Iron
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