| Question | Answer |
| 3 classifications of carbs | Monosaccharides,
oligosaccharides,
polysaccharides |
| What is the only monosaccharide found in the body? | Glycogen |
| Monosaccharides and oligosaccharides are_ | Soluble in aq. solution,
difficult to demo |
| 4 groups of polysaccharides | Group I: neutral polysaccharides
Group II: acid mucopolysaccharides
Group III: glycoproteins
Group IV: glycolipids |
| Group I includes_ | glycogen, starch, cellulose, chitin,
PAS-positive |
| Group II includes_ | Hyaluronic acid, heparin,
chondroitin sulfates A, B, & C,
human aorta, bovine cornea,
"connective tissue mucins,"
PAS-negative |
| Group III includes_ | ovimucoid, mucin,
Paneth cell granules,
sialomucins, sialogycoproteins,
serum glycoproteins,
"epithelial mucins,"
maybe PAS-positive |
| Group IV includes_ | Cerebrosides: fatty subs. w/ carb,
phosphatides: non-carb lipids,
PAS positive |
| PAS purpose | Demo polysaccharides,
neutral mucosubstances,
basement membranes |
| PAS facts | 10% NBF or Bouin;
blood smear - methanol,
kidney 1-2 μm,
other 4-5μm,
QC: kidney; demo glycogen - liver or cervix |
| PAS solutions | Periodic acid +
Schiff Reagent +
potassium metabisulfite +
Harris w/ acetic acid or light green |
| PAS results | Positive reaction: bright rose -
nuclei: blue if hematoxylin -
background: light green if counterstained |
| Schiff reagent solution | sodium metabisulfite + HCL = sulfurous acid +
basic fuchsin +
activated charcoal +
dH2O
reacts w/ aldehydes to restore color structure |
| What substances give positive PAS reaction? | Glycogen, neutral mucosubstances,
some epithelial sulfomucins & sialomucins,
colloid material of thyroid,
pars intermedia of pituitary,
basement membranes, fungal walls |
| PAS w/ diastase digestion purpose | Demo glycogen |
| PAS w/ diastase digestion facts | 10% NBF, formalin alcohol, or absolute alcohol
QC: 2 sections of liver or cervix w/ glycogen - label one "w/," one "w/o" |
| PAS w/ diastase digestion solutions | malt diastase solution +
periodic acid +
Schiff reagent +
potassium metabisulfite +
Harris w/ acetic acid |
| PAS w/ diastase digestion results | Glycogen on "w/o" section: bright rose red -
glycogen on "w/" section: no stain |
| Malt Diastase solution | diastase of malt +
phosphate buffer, pH 6 |
| Phosphate buffer, pH6 | NaCl +
sodium phosphate +
H2O |
| Mayer mucicarmine purpose | Demo "epithelial" mucin |
| Mayer mucicarmine facts | 10% NBF
QC: small intestine, appendix, or colon |
| Mayer mucicarmine solutions | Mucicarmine solution +
Weigert hematoxylin +
Metanil yellow or light green +
H2O |
| Mayer mucicarmine results | epithelial mucin, capsule of cryptococcus: deep rose to red -
nuclei: black -
other: blue or yellow |
| Mayer mucicarmine stock solution | carmine + alum lake +
aluminum hydroxide + anhy. aluminum chloride +
50% ethanol +
25% ethanol |
| Metanil yellow solution | metanil yellow +
acetic acid +
H2O |
| Alcian blue, pH 2.5 purpose | demo acid mucopolysaccharides |
| Alcian blue, pH 2.5 facts | 10% NBF or Bouin
QC: small intestine, appendix, or colon positive control |
| Alcian blue, pH 2.5 solutions | acetic acid +
alcian blue +
nuclear-fast red |
| Alcian blue, pH 2.5 results | acid mucopolysaccharides: dark blue -
background/other: pink to red |
| Nuclear-fast red (Kernechtrot) solution | nuclear-fast red +
aluminum sulfate +
H2O |
| Alcian blue, pH 1.0 purpose | demo sulfated mucosubstances |
| Alcian blue, pH 1.0 facts | 10% NBF or Bouin
QC: small intestine, appendix, or colon positive control |
| Alcian blue, pH 1.0 solutions | HCl +
alcian blue, pH 1 +
Nuclear-fast red |
| Alcian blue, pH 1.0 results | sulfated mucosubstances: pale blue -
background: pink to red |
| Alcian blue w/ hyaluronidase purpose | Differentiate epithelial & connective tissue mucins,
hyaluronidase digests out connective mucins |
| Alcian blue w/ hyaluronidase facts | 10% NBF
QC: 2 sections of umbilical cord - one labeled "w," one "w/o,"
may use small intestine, appendix, or colon as 2nd control for epithelial mucins |
| Alcian blue w/ hyaluronidase solutions | hyaluronidase digestion soln. +
acetic acid soln. +
alcian blue +
nuclear-fast red |
| Alcian blue w/ hyaluronidase results | w/o, acid mucopolysaccharides, sialomucins: deep blue -
w/, mucosubstances w/ hyaluronic acid, chondroitin sulfates A & C: marked loss of staining |
| Buffer solution, pH 6.0 | potassium phosphate +
sodium phosphate |
| Hyaluronidase digestion solution | testicular hyaluronidase +
buffer soln. |
| Alcian blue-PAS-hematoxylin purpose | Differentiate between neutral & acidic mucosubstances |
| Alcian blue-PAS-hematoxylin facts | 10% NBF, Zenker,
kidney 2-3 μm,
other 4-5μm
QC: kidney or mucin control depending on diagnostic tissue |
| Alcian blue-PAS-hematoxylin solutions | acetic acid soln. +
alcian blue, pH 2.5 +
periodic acid soln. +
Schiff reagent +
reducing rinse +
Harris w/ acetic acid |
| Alcian blue-PAS-hematoxylin results | acid mucosubstances: blue -
neutral polysaccharides: magenta -
substances stained by PAS & alcian blue: purple |
| Reducing rinse solution | sodium metabisulfite +
H2O
prepare just before use |
| Müller-Mowry colloidal iron purpose | Demo carboxylated & sulfated, (acid) mucopolysaccharides
& glycoproteins,
less specific than alcian blue,
Prussian blue reaction |
| Müller-Mowry colloidal iron facts | 10% NBF, Carnoy, alcoholic formalin;
avoid chromate fixatives
QC: small intestine, appendix, or colon |
| Müller-Mowry colloidal iron solutions | acetic acid soln. +
Müller-Mowry colloidal iron +
ferrocyanide-HCl +
Nuclear-fast red |
| Müller-Mowry colloidal iron results | acid mucopolysaccharides & sialomucins: deep blue -
nuclei: pink-red -
cytoplasm: pink |
| Müller-Mowry colloidal iron working solution | ferric chloride +
acetic acid +
H2O |
| Ferrocyanide-hydrochloric acid solution | potassium ferrocyanide +
HCl +
H2O |
| What is amyloid? | Fibrillar protein,
deposits in tissue under pathologic conditions,
contains some acid mucopolysaccharides |
| What is amyloidosis? | Disease where deposits replace cellular elements causing
progressive loss of function & eventual death |
| Primary amyloid | Occurs in absence of disease,
primarily muscle, heart, skin, tongue |
| Secondary amyloid | Associated w/ disease, esp. inflammatory disease, e.g. RA & TB,
primarily kidneys, liver, spleen, adrenal glands |
| Myeloma-associated amyloid | Associated w/ diseases of immunological system,
primarily muscle, heart, skin, tongue |
| Tumor-associated amyloid | Associated w/ tumors,
esp. those of amine precursor uptake & decarboxylation system |
| Alkaline Congo-red method facts | Demo amyloid,
alcohol or Carnoy
8-10 μm (or no birefringence)
QC: sections w/ amyloid (brain, kidney)
stain intensity decr. over time |
| Alkaline Congo-red method solutions | Harris w/ acetic acid +
alkaline salt soln. +
Congo red working soln. |
| Alkaline Congo-red method results | amyloid: deep pink to red -
elastic tissue: pale pink -
nuclei: blue |
| Alkaline salt solution | 80% alcohol sat'd w/ NaCl +
NaOH |
| Congo red working solution | Congo red +
80% alcohol sat'd w/ NaCl +
NaOH |
| Stock 80% alcohol sat'd w/ NaCl | NaCl +
100% ethanol +
H2O |
| Crystal violet purpose | Rapid screening of amyloid,
not as specific as Congo red |
| Crystal violet facts | 10% NBF or alcohol
10-12 μm
QC: section w/ amyloid
polychromatic |
| Crystal violet solutions | 95% sat'd w/ CV +
Apathy mounting medium,
seal w/ nail polish |
| Crystal violet results | amyloid: purplish violet -
other elements: blue |
| Modified Apathy mounting medium solution | gum arabic +
cane sugar +
NaCl +
thymol +
H2O |
| Thioflavin T Fluorescent method facts | Demo amyloid, less specific than Congo red
10% NBF
6-10 μm
QC: section w/ amyloid |
| Thioflavin T Fluorescent method solutions | Mayer +
thioflavine T +
acetic acid soln. |
| Thioflavin T Fluorescent method results | amyloid: fluoresces yellow to yellow-green |
| How to test Schiff reagent quality? | Schiff reagent should be clear/light amber;
to test quality add formaldehyde,
reddish-purple - good,
deep blue-purple - solution breaking down |
| Best carmine facts | Demo glycogen, less specific,
alcohol, Bouin, Carnoy |
| Best carmine reagents | carmine +
K2CO3 + KCl
NH4OH +
methanol +
ethanol +
hematoxylin |
| Best carmine results | glycogen: pink -
nuclei: blue |
| Alcian blue is a: | Copper phthalocyanin dye |
| Prussian blue reaction: | Acidic potassium ferrocyanide,
ferric iron reacts to form insoluble bright blue pigment |
| Congo red birefringence under polarized light should be: | Bright green, 8-10 µm,
too thin - red,
too thick - yellow |
| Turnball blue reaction: | Acidic potassium ferrocyanide,
ferrous iron reacts to form insoluble bright blue pigment - ferrous ferricyanide |
| The colored product of the PAS reaction is formed by: | Restoration of the quinoid structure |
