Module 1: Staining terms Flashcards
Natural dye derived from Heartwood of logwood tree and is used for nuclear staining
Hematoxylin
Natural dye derived from Cochineal bug and is used for glycogen staining
Carmine
Natural dye derived from Dried stigmus of crocus flower extracted at 56DegC and is used for collagen staining
Saffron
Natural dye derived from Derived from Lichens and is used for elastic fiber staining
Orcein
Natural dye derived from derived from lichens and is used as a pH indicator
Litmus
Synthetic dyes
- Usually derivatives of benzene
- colorless in visible spectrum so needs to be altered to absorb light in visible spectrum by addition of elements to benzene ring (chromophores)
Chromophore
The more chromophore, the more intense the color
Easily reduced due to affinity for hydrogen
Most common groups are Quioninoid, Nitro and Azo
Chromagen
Benzene + chromophore
- not a dye, no affinity for fibers or tissues
- ionizing group (auzochrome) is added to provide affinity
Auxochrome
- allows chromagen to ionize and link with tissue structure
- attached to chromophore portion of dye molecule
- enable chromgen to attach itself to tissue substances of opposite charge OR act as color modifier
- Can have acidic or basic charges when they ionize
Acidic auxochromes
COOH (carboxyl) ionizes to COO-
OH (hydroxyl) ionzes to O- (phenolic-associated with aromatic ring)
SO3H (sulphonic) ionizes to SO3- (not a true auxo, needs another one present to be functional)
Basic auxochromes
NH2 (amino)
total dye complex (benzene + chromophore + auxochrome =)
auxochromophoric system
auxochrome actions
- Amine, carboxyl and phenolic are FUNCTIONAL
- Sulphonic is salt forming group that strongly acidic and isn’t able to form union with tissue alone. Makes dye soluble in water and changes cationic dye to an anionic dye
- If both acid and base auxochromes are present, amine predominates over carboxyl and hydroxyl (but weakened)
- Sulphonic group predominates over amine groups
Modifiers
chemical end groups that alfter color or intensity of dye (Ex. methyl, aryl, ethyl)
Chemical classification of dyes by Charge
Acidic/anionic dyes
Basic/cationic dyes
Neutral dyes
Amphoteric dyes
Chemical classification of dyes by Charge: Acidic/anionic dyes
net negative charge on chromagen.
Salt of sodium
Act as acid in solution
Attaches to basic tissue groups (acidiphilic tissues)
Chemical classification of dyes by Charge: Basic/cationic dyes
Net positive charge
Salt of chloride
Act as base in solution
Attaches to basophilic tissues
Chemical classification of dyes by Charge: Neutral dyes
Both sodium and chloride salt are mixed.
Soluble in alcohol, but rarely in water
Chemical classification of dyes by Charge: Amphoteric dyes
Cationic below a certain pH (isoelectric point) and anionic above it.
When dye is in acid solution, it behaves as a basic dye, and vice versa.
Acidic tissue components
RNA
DNA
Cartilage matrix
Many mucus secretions
- basophilic tissue (affinity for basic stain)
- anionic/acidic charge (-)
Basic tissue components
Collagen
RBC cytoplasm
Granules of eosinophilic leukocytes
Acidophilic tissue (affinity for acid stain) -cationic/basic charge (+)
Amphoteric tissue components
Cytoplasm of most cells
Contractile elements of muscle
- decreased pH make net charge (+)
- increased pH make net charge (-)
Neutrophilic tissues components
Neutrophilic granules of leukocytes
-Structures with affinity for neutral or compound stain as complete or nonionized molecule
Dye Nomenclature (CSMLS)
capitalize first letter of all proper names and letter shade designation
QC for dyes and dye solutions
- Mark with date of receipt and store in cool place away from light in airtight container
- In solid form, will keep for years
- QV slides must be checked before sending to pathologist
Stain prep
- ensure color index #, dye lot#, and purity on label match whats required for method
- note conditions (pH, temp) and storage conditions
- Some dyes require certain solvents
- Mark stain with name, date, initials
Storage and reuse of dyes
store RT, away from light in airtight container
-Discard once shows precipitate or gives poor staining results
Stains requiring refrigeration
Schiffs reagent Bests' carmine Aldehyde fuschion Methyl green Azocarmine Stock solutions of methenamine silver nitrate