Carbohydrate and Amyloid

Principle/Purpose

100

Oxidation of tissue elements to aldehydes by periodic acid then letting Schiff happen.

What is PAS?

200

Differentiate connective tissue mucins from epithelial tissue mucins

Alcian blue with hyaluronidase

300

Tissue treated with ferric hydroxide in suspension then placed in ferrocyanide-hydrochloric acid solution to stain acid mucins deep blue.

Colloidal iron

400

Rapid screening of amyloid but not as specific as Congo red. Is considered polychromatic with amyloid staining purple-volet and other tissue elements staining blue

Crystal violet

500

Copper phthalo cyanic basic dye that is water soluble 

Alcian blue

Reagents

100

Periodic acid, schiff reagent , potassium metabisulfite

PAS

200

Alcian blue, hydrochloric acid, nuclear fast red

Alcian blue 1.0

300

Alcian blue, acetic acid, Kernechtrot solution

Alcian blue 2.5

400

Basic fuchsin, sodium metabisulfite, hydrochloric acid, water

Schiff reagent

500

Metanil yellow, Weigert hematoxylin, carmine

Mucicarmine

Results

100

Amyloid......deep pink to red

Elastic tissue....pale pink

Nuclei.....Blue

Congo red

200

Exclusively acid mucosubstances......Blue

Neutral polysaccharides......Magenta

Mixed mucins........Purple

AB/PAS

300

Amyloid is stained fluoresces yellow to yellow-green

Thioflavin T

400

Counterstain, must be washed well after use or clouding will result once placed in alcohol

Nuclear fast red

500

Mucin....deep rose to red

Capsule of cryptococcus neoformans....deep rose to red

Nuclei....black

Other tissue elements.....blue or yellow

Mayer mucicarmine

Procedure

100

Place sections in alkaline salt solution for 20 minutes

Congo red

200

Stain two slides one with digestion and one without

PAS W/ diasase

300

Rinse sections briefly in 0.1N HCL

Alcian blue 1.0

400

Immerse in ferrocyanide-hydrochloric acid solution for 20 minutes at room temperature

Muller colloidal iron

500

Wash in running tap water for 10 minutes to develop full color. Neutral mucins only

PAS

Technical Notes

100

This method is excellent for the demonstration of Cryptococcus neoformans

Colloidal Iron

200

Used in place of hematoxylin when demonstrating fungus

Fast green or Light green

300

Fixative that will cause a false positive reaction with PAS staining duo to it being a di aldehyde

Glutaraldehyde

400

Counterstain, must be washed well after use or clouding will result once placed in alcohol

Nuclear Fast red

500

Intracellular secretions of various cells that stain with basic dyes, metachromatic, soluble in alkaline solution, some precipitated by acetic acid.

Mucins