Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Microtechniques > Carbohydrates CH 7 > Flashcards

Carbohydrates CH 7 Flashcards

1
Q

Single most abundant class of organic molecules in nature.

The metabolic precursors
of virtually all other biomolecules
Formula: (CH₂O)n
Including: Sugars, starch, cellulose…

A

Carbohydrates

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Major saccharides of concern

A

Glucose

Glycogen (storage form of glucose)

Additional types of concern: Homoglycans, heteroglycans, mucins, glycolipids

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

1.) Type of saccharide is Glucose

2.) Type of saccharide is Glycogen (storage form of glucose)

A

1.) Monosaccharide

2.) Polysaccharide

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Carbohydrate Pathological Conditions
1.) Name the carbohydrate metabolism enzyme deficiencies.

2.) Glycogen may have diagnostic significance in
several types of tumors including what?

A

1.)
Von Gierke’s disease
Pompe’s disease

2.)
Carcinoma
Mesothelioma
Rhabdomyosarcoma

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Carbohydrate Pathological Conditions
◦ Mucins function as lubricants or assist in cell adhesion or host defense
◦ Can be from epithelial or connective tissue origin

What are mucins produced by?

A

By many tumors including
- Carcinoma
- Liposarcoma
- Mesothelioma

Enzyme deficiencies can cause abnormal systemic production of mucins
- Hurler syndrome
- Hunter disease

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

For histopathology concerns, there are four main groups of carbohydrates for demonstration. They are:

A
  1. Neutral Polysaccharides (Nonionic homoglycans)
  2. Acid Mucopolysaccharides (Anionic heteroglycans also known as glycosaminoglycans)
  3. Glycoproteins (Mucins, Mucoid, Mucoproteins,
    mucosubstances)
  4. Glycolipids
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Neutral Polysaccharides (Nonionic homoglycans) group

A
  • Either glucose containing
  • Like glycogen, starch, cellulose
  • Or N-acetyl-glucosamine containing
  • Like chitin
  • Group is VERY positive PAS
  • Negative with other common carbohydrate stains
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

Acid Mucopolysaccharides (Anionic heteroglycans) group

A
  • Either carboxylated: like hyaluronic acid
  • Or chondroitin sulfated: Connective tissue compounds
  • Or heparin (mast cells); sulfated and carboxylated;
  • Sulfated only: Aorta
  • Group is considered proteoglycans (attached to protein)
  • PAS negative. Positive with other common carb stains
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Glycoproteins (mucins, mucoid, mucoprotein, mucosubstances) group

A
  • Either neutral: Stomach mucins, Paneth cell granules
  • Or carboxylate, sialomucins: goblet cells, serum, blood or sulfated and carboxylated groups
  • Group is mostly epithelial mucins
  • Potentially but not always PAS positive. Demonstrated
    with other stains or techniques
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Glycolipids group

A
  • Either cerebrosides: fatty residue bound to a carbohydrate structure
  • Phosphatides: PAS positive noncarbohydrate containing lipids
  • Not commonly demonstrated in clinical laboratory
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

Carbohydrate Special Stains

A

1.) PAS (Periodic acid, Schiff): PAS with Diastase
2.) Best Carmine
3.) Mayer Mucicarmine
4.) Alcian Blue, pH 2.5
- Alcian Blue, pH 1.0
- Alcian Blue with Hyaluronidase
- Alcian Blue-PAS-Hematoxylin
5.) Colloidal Iron

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

PAS (Periodic Acid Schiff) Reaction

A
  • Demonstrates polysaccharides, neutral mucosubstances, basement membranes
    (measure invasiveness of skin tumors) and fungus (fungal wall)
  • Sections cut at 4 to 5 microns (1 to 2 microns for kidney)
  • Control tissue: kidney or small or large intestine (liver to demonstrate glycogen only)
  • Fixative: 10% NBF or Bouin solution
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

PAS (Periodic Acid Schiff) Reaction

A
  • Reaction is based on oxidation of certain tissue elements to aldehydes by periodic acid.
  • Schiff’s is compound of basic fuchsin and sulfurous acid (forming a colorless compound)
  • Schiffs reaction with aldehydes (attachment of dye structure), followed by washing in running water, causes the attached dye color to become visual by breaking the sulfurous group away from basic fuchsin
  • Results: PAS positive for neutral mucosubstances, glycogen, basement membranes, fungal walls – BRIGHT ROSE/MAGENTA
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

PAS (Periodic Acid Schiff) Reaction Typical Stain Protocol

A
  • Periodic acid solution
  • Rinse in running water
  • Schiff reagent
  • Rinse in metabisulfite**
  • Running tap water for 5 to 10 minutes
  • Counterstain with hematoxylin
  • Metabisulfite rinses may be used to remove excess Schiff reagent, preventing false staining of oxidized absorbed reagents (i.e. highly chlorinated water). Must follow with running water after sulfite rinses.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

PAS Reaction Troubleshooting

A
  • Glutaraldehyde can NOT be used (di-aldehyde)
  • Open aldehyde stains
  • PAS false positive
  • Chromate containing fixatives may “over- oxidize” tissue
  • Schiff reagent is typically stored between 2ºC to 12ºC.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

PAS with Diastase Digestion
- Demonstration of glycogen

A
  • 2 slides are prepared:
    1.) First slide WITH enzyme diastase (or α-amylase) digests glycogen from tissue section

2.) Second slide of same tissue WITHOUT diastase treatment

  • Diastase digestion shows “where glycogen was” when compared to slide WITHOUT diastase (otherwise, it was not glycogen).
  • Fixative: 10% NBF, formalin alcohol, absolute alcohol
  • Avoid picric acid fixatives; glycogen is more resistant to digestion
17
Q

PAS with Diastase Digestion
- Typical Staining Protocol

A
  • Section WITH Diastase, 1 hr at 37ºC
  • Running water 5 minutes
  • Both sections in Periodic acid
  • Continue as with standard PAS protocol
18
Q

Best Carmine
- Demonstration of glycogen

A
  • Sections cut at 4 to 5 microns
  • Control tissue: liver
  • Results: Glycogen-Pink to Red; Nuclei-Blue
  • Fixative: Absolute alcohol, Carnoy solution, Bouin solution
19
Q

Best Carmine
- Typical Staining Protocol

A
  • Harris hematoxylin
  • Running water 15 minutes
  • Working Carmine solution 30 minutes
  • Differentiate 30 seconds
  • Rinse quickly 80% alcohol
  • Run Down
  • Not as specific as PAS with Diastase.
  • Ammonium hydroxide component toxic, corrosive
20
Q

Mucicarmine
- Demonstration of epithelial mucins

A
  • Sections cut at 4 to 5 microns
  • Control tissue: Small or large intestine, appendix
  • Results: Mucin-Deep Rose to Red; Nuclei- Black; Cryptococcus neoformans-Deep Rose to Red
  • Fixative: 10% NBF
21
Q

Mucicarmine
- Typical Staining Protocol

A
  • Weigert hematoxylin
  • Running water
  • Working Mucicarmine solution (alum Carmine)
  • Rinse and remove water quickly
  • Mentanil yellow
  • Run down
22
Q

Alcian Blue pH 2.5
- Demonstrates acid mucopolysaccharies

A
  • Section at 4 to 5 microns
  • Control Tissue: small or large intestine, appendix
  • Results: acid mucosubstances-DARK BLUE, background-PINK TO RED,
  • Fixative: 10% NBF or Bouin solution
23
Q

Alcian Blue pH 2.5
- Typical Staining Protocol

A
  • 3% acetic acid solution
  • Alcian blue solution for 15 minutes at 37ºC
  • Rinse in 3% acetic acid
  • Running water for 10 minutes
  • Counterstain with nuclear-fast red for 5 minutes
  • Copper based dye that is colored blue due to copper content
24
Q

Alcian Blue pH 1.0
- Demonstrates sulfated mucosubtances

A
  • Section at 4 to 5 microns
  • Control Tissue: small or large intestine, appendix
  • Results: sulfated mucosubstances-PALE
    BLUE, background-PINK TO RED,
  • Fixative: 10% NBF or Bouin solution
25
Q

Alcian Blue pH 1.0
- Typical Staining Protocol

A
  • 0.1N HCl solution (appx 1% HCl)
  • 1% alcian blue solution for 15 minutes at 37ºC
  • Rinse in 0.1N HCl solution
  • Blot dry (to avoid pH change from water)
  • Counterstain with nuclear-fast red for 5 minutes
  • Rinse in water then run down as normal
26
Q

Alcian Blue with Hyaluronidase
- Differentiation of epithelial and connective tissue mucins

A
  • 2 slides are prepared:
    1.) First slide WITH enzyme Hyaluronidase to digest connective tissue mucins from tissue section

2.) Second slide of same tissue WITHOUT Hyaluronidase treatment

  • Digestion shows “where connective tissue mucin was” when compared to slide WITHOUT digestion.
  • Results: WITHOUT digestion acid mucopolysaccharides-DARK BLUE, WITH digestion shows loss of staining (absence of hyaluronic acid) background-PINK TO RED
  • Fixative: 10% NBF
27
Q

Alcian Blue-PASH
- Differentiation between neutral and acidic
mucosubstances

A
  • Section at 4 to 5 microns
  • Control Tissue: kidney or mucin control. Cervix section (both endocervix and ectocervix)
  • Results: acidic mucosubstances-DARK BLUE, neutral polysaccharides-Magenta, background-PINK TO
    RED
  • Fixative: 10% NBF or Zenker solution
28
Q

Alcian Blue-PASH
- Typical Staining Procedure

A
  • 3% acetic acid
  • Alcian blue
  • Running tap water
  • Periodic acid
  • Running tap water
  • Schiff reagent
  • Running tap water
  • Hematoxyilyn
  • Run down
29
Q

Colloidal Iron
- Demonstration of carboxylated and sulfated
mucopolysaccharides and glycoproteins
.

A
  • Section at 4 to 5 microns
  • Control Tissue: small or large intestine, appendix
  • Results: Acid mucopolysaccharides and sialomucins- Deep blue, Nuclei-Pink to Red, Cytoplasm-Pink
  • Fixative: 10% NBF, Carnoy solution or alcoholic
    formalin. Avoid chromate fixatives.
30
Q

Colloidal Iron
- Typical Staining Protocol

A
  • 12% acetic acid
  • Working Colloidal Iron
  • 12% acetic acid
  • Ferrocyanide-hydrochloric acid
  • Running water
  • Nuclear-fast red counterstain
  • Can also be used for Cryptococcus neoformans
31
Q

Amyloid

A
  • Intercellular material that is deposited in various tissues such as: Heart, Muscle, Skin, Liver, Spleen, Kidneys and Brain
  • Originally considered carbohydrate, but is a structure based on fibril subunit proteins
  • Replaces vital tissue, causing organ failure
  • Clinical amyloidosis associated with: Genetic predispostion, chronic inflammatory diseases, tumors, and alzheimer disease
32
Q

Congo Red
- Demonstration of amyloid in tissues

A
  • Section at 8 to 10 microns
  • Control Tissue: anything with amyloid
  • Results: Amyloid-Deep pink to red
  • Fixative: 10% NBF, Carnoy solution or alcoholic formalin. Avoid chromate fixatives.
33
Q

Congo Red
- Typical Staining Protocol

A
  • Harris hematoxylin
  • Running water
  • Alkaline salt solution
  • Congo red staining solution
  • Run down
  • With polarizing microscope, apple-green birefringence is demonstrated by amyloid (not glycogen)
  • Preferred method for amyloid
34
Q

Crystal Violet
- Demonstration of amyloid in tissues

A
  • Section at 10 to 12 microns
  • Control Tissue: anything with amyloid
  • Results: Amyloid-Purplish violet
  • Fixative: 10% NBF or alcohol
35
Q

Crystal Violet
- Typical Staining Protocol

A
  • Crystal violet solution
  • Running water
  • Air dry, dip in xylene, mount
36
Q

Thioflavin T Fluorescent method
- Demonstration of amyloid in tissues

A
  • Section at 6 to 10 microns
  • Control Tissue: anything with amyloid
  • Results: Amyloid-Fluoresces yellow (hence the name)
  • Fixative: 10% NBF
37
Q

Thioflavin T Fluorescent method
- Typical Staining Protocol

A
  • Mayer hematoxylin
  • Stain in thioflavin T
  • Rinse in DI
  • Differentiate in 1% acetic acid
  • Wash in running
  • Thioflavin T is a fluorescent dye that attaches to amyloid
  • Hematoxylin decreases background nuclear staining
  • Must use a fluorescent microscope to view

Microtechniques (10 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions