CARBOHYDRATE STAINS

Carbohydrates
main sources of energy
forms monosaccharide, polysaccharides
glycogen - stored form of CHO in humans and animals
plant form: STARCH
stored in liver

Mucin
polysaccharides bound to other substances
forms the matrix of connective tissues; secreted by goblet cells, respiratory lining cells, certain
glands
found in epithelial and intestinal glands
classified as:
Mucopolysaccharides
Acid Mucopolysaccharides
USE Alcian Blue
primarily composed of hyaluronic acid, heparin sulfate and chondroitin sulfate
metachromatic substances
Periodic Acid Schif (NEGATIVE)
Mucoproteins (mucoids and glycoproteins)
Neutral Mucopolysaccharides
stained with PAS
contains hexoses
do not stain with Alcian Blue
Stain Use Color Reaction Unique Feature

Periodic Acid Schif stains: Magenta Intensity of PAS

glycogen proportional to sugar
mucoproteins - most content
common PAS (+)
hyaline
casts
glomerular basement
membrane

PAS with Diastase method of choice for Red Control - only nuclei addition of Diastase
Control staining glycogen is stained serves as a negative
negative control enzyme control

Best Carmine glycogen Bright Red Selective and Highly

Specific for Glycogen
Langhans Iodine oldest method for Mahogany Brown
Method glycogen; already
obselete

Stains for Mucin

Alcian Blue Acid Blue most popular method

Mucopolysaccharide

Azure A Acid Crimson or Red Violet most useful

Mucopolysaccharide metachromatic dye
Fixative: Mercuric
Chloride

Uranyl Acetate Azure A Acid MPS Crimson or Red Violet metachromatic stain

Toluidine Blue Acid MPS Red Purple metachromatic stain

Combined Alcian Blue Acid and Neutral Mucin Acid Mucin - Blue Separates Acid and
Pas Technique Neutral Mucin - Magenta Neutral Mucins

Combined Gomoris Mucins Sulfated Mucin - Purple greater affinity for

Aldehyde Fuchsin - Carboxylated - Blue sulfated mucin
Alcian Blue

Mucicarmine Mucins Red addition of Aluminum

Hydroxide Southgates
Mucicarmine
Technique -
Cryptococcus
neoformans

Hales Colloidal Mucins Acid Mucin - Dark Blue Greater sensitivity and
(Dialyzed) Iron intensity compared to
Technique alcian blue

Fluorescent Acridine Acid Mucins Brilliant Orange Uses Fluorochrome

Orange Technique Fluoresence Stains temporarily

STAINS FOR PROTEINS, ENZYMES AND NUCLEIC ACIDS

Proteins
made up of amino acids and are linked by peptide bonds
Histochemical Identification of Proteins
demonstrates presence of amino acids
 Neutral Bufer Formol Saline - most commonly used fixative
Enzyme Histochemistry
detection of enzyme which act as a biological catalyst
tissues used:
frozen section (-70C or below) fixed in acetone or formalin
fresh frozen section
tissue fixed in 4% formalin or formol saline overnight
Metal Precipitation - most common technique for histochemical demo of enzymes

Nucleic Acid
composed of alternate sugar and phosphate groups with a nitrogenous base
two types: RNA and DNA
usually combined with basic proteins to form nucleoproteins
CONNECTIVE TISSUE STAINING

Stain Use Color Reaction Unique Feature

Alkaline Fast Green Histones and Protamines Green Histochemical ID of

Protein

Peracetic Acid Alcian Cystine and Cysteine Blue Green Histochemical ID of

Blue Amino Acids

Sakaguchis Arginine Orange Red Histochemical ID of

Amino Acids

Enzyme Histochemistry *Only those highlighted by sir

Metal Precipitation ATPase Light Brown/Dark Brown

Feulgen Technique Nuclear DNA Red Purple most reliable and specific

primarily stain Connective Tissue fibers

CT fibers - produced by fibroblast
3 types:
collagen , elastic , reticular

STAINS FOR PATHOLOGIC CHANGES AND DEPOSITS FOUND IN CONNECTIVE TISSUES

1. Fibrin
insoluble fibrillar protein
seen after tissue damage, blood clots, acute inflammatory reactions
2. Fibrinoid
eosinophilic
identical staining reactions to fibrin
seen in collagen diseases, hypersensitivity, SLE
3. Hyalin

Stain *Only those Use Color Reaction Unique Feature

highlighted by sir

Gomoris Silver Reticulin - always Black

Impregnation

wide variety of pathologic exudates and deposits

stain: non-specifically PAS
4. Amyloid
semi-translucent, ground glass or hyaline eosinophilic substance
composed of chondroitin sulfuric acid protein complex
deposited in CT cells, kidney, spleen, adrenals, etc.

Stain *Only those Use Color Reaction Unique Feature

highlighted by sir

Congo Red Amyloid Red alkaline congo red -

method of choice