Carbohydrate Stains - PAS, Alcian Blue, Toluidine Blue (Wk 7) Flashcards
This is the property of certain dyes which stain different colours depending on what tissue element they are attached to.
METACHROMASIA
Schiff’s effectiveness can be tested by adding a few drops to ________
formalin
This is the term for alcohol groups on adjacent carbons in a sugar molecule?
VICINAL DIOLS
This is what the D in PAS + D stands for.
DIASTASE
This value is one measurement of acid strength and is useful in determining at which pH anionic components will be ionized.
pKa
Schiff’s reagent binds to this reactive group
ALDEHYDES
3 important factors in metachromatic staining
- pH
- Water (dehydration will destroy metachromatic staining)
- Spacing between anionic groups (0.3-0.5 nm spacing between groups; otherwise won’t stain)
Your spit contains this enzyme.
AMYLASE
This is one of the major types of mucosubstances and is found in connective tissues?
PROTEOGLYCANS
2 components in Mucicarmine?
aluminum + carmine
This is the name for an anionic group which will stain red-purple with Toluidine blue.
CHROMOTROPE
Schiff’s reagent may revert back to pararosaniline which is what colour?
pink
3 alternate stains for Alcian blue
- Mucicarmine
- Colloidal iron
- Lectins
The carbohydrate component of a proteoglycan is called?
Glycosaminoglycan
Glycoproteins can be removed by using..?
Neuraminidase
Why must Alcian blue/PAS be done in a particular order, and what that order is?
Alcian before PAS
Schiff’s will introduce sulphonic acid groups = false positives if stained w/ Alcian
Chromophore in pararosaniline?
Quinoid ring
When this kind of a chemical group is ionized it can be stained with Alcian blue at pH 2.5 but not at pH 1.0?
CARBOXYLIC ACID
2 types of carbohydrates that stain with Alcian Blue
Proteoglycans
Acid glycoproteins (NOT neutral)
The name for HIO4
PERIODIC ACID
This is the most common dye used to demonstrate amyloid.
CONGRO RED
Proteoglycans can be removed by using..?
Hyaluronidase
These mucosubstances are considered to be epithelial mucins.
GLYCOPROTEINS
Why is pararosaniline a PSEUDOLEUCO dye?
Colourless (due to addition of SO3- to chromophore) and colour is re-established in presence of aldehydes
Not a true leuco dye b/c doesn’t involve redox rxn
Why is pararosaniline preferred over basic fuchsin to make Schiff’s reagent
Basic fuchsin is a MIXTURE of dyes
Pararosaniline is a SINGLE dye (better specificity)
General description of dyes which can be used to demonstrate acid mucins and other negatively charged components?
CATIONIC
This is the pH at which only sulphated proteoglycans will stain.
pH 1.0
Modern Schiff’s reagent is made from this single dye
Pararosaniline
Repetitive anionic groups responsible for metachromatic staining
Chromotropes
Purpose of staining at pH 1.0 and 2.5 for Alcian blue?
Differentiate between sulphate and carboxylated mucins (via none/increase in staining intensity)
COOH only stains at 2.5
This dye will specifically demonstrate Cryptococcus neoformans.
MUCICARMINE
This simple sugar is rarely seen in human tissues due to its solubility.
GLUCOSE
What functional group gets attached to carbohydrates after the addition of Schiff’s reagent?
SO3H
PAS is an example of a staining method that works via __________ bonding
covalent
2 potential false positives in PAS due to the oxidation step?
- Glutaraldehyde-fixed tissues
- Some lipids
The liver would be one place to find this mucosubstance.
GLYCOGEN
PAS stain will demonstrate neutral mucins located in ______________ of liver
portal triads
This is a mucosubstance which is often linked to proteins in glycoproteins?
SIALIC ACID
Used to detect aldehydes?
SCHIFFS REAGENT
How is the chromophore in pararosaniline modified?
Destroyed by addition of SO3- to form colourless solution
4 main PAS(+) substances
- Glycogen
- Neutral glycoproteins (stomach)
- Neutral glycoproteins (SI)
- Basement membranes
Why don’t proteoglycans stain PAS-positive?
Periodate ion (HIO4-) is repelled by COO- and SO3- parts of proteoglycan = no oxidation
Oxidizing agent in PAS stain
Periodic acid
This is a term for anisotrophic substances.
BIREFRINGENT