BASIC

PRINCIPLES OF
STAINING
STAINING
Staining always involves the visual labeling of some biological
entity by attaching, or depositing in its vicinity, a marker of
characteristic color or form.
Is the process of coloring cells, cellular constituent & tissue
fibers to facilitate optical differentiation by microscopic
examination.
•Is the union between a colored dye & a tissue substrate
which resists simple washing.
STAIN
Stain is defined as “any chemical substance which when added
to the living cells or to fixed structures or structural
components, makes them clearly visible or detectable.”
A stain is the marker, or the reagent used to generate the marker.
 Outlines tissues and cellular components.

 Identification of tissues.

 Establishes the presence or absence of

disease processes.
Classification of dyes
CLASSIFICATION I
Basic dyes
Acidic dyes
Neutral dyes
CLASSIFICATION 2
Natural dyes
Synthetic dyes
BASIC DYES
Basic dyes are cationic and will stain anionic or acidic
materials such as carboxylates, sulphates and phosphates.
Most are used as nuclear stains.
Acidic substances that stain with basic dyes are termed
basophilic.
BASIC:
Haematoxylin
Acridine red
Aniline blue
Azure
Basic Fuschin
Crystal violet
ACIDIC DYES
Acidic dyes are anionic and will stain positive charged groups
in tissue
Most are used to stain proteins in the cytoplasm and connective
tissues.
Substances that stain with acid dyes are called acidophilic.
 
ACIDIC:
Eosin
Erythrosine
Picric acid
Alizarin
Acid fuschin
Bismarck brown
 
NEUTRAL DYES
Neutral dyes are simply compounds of basic and acid dyes.
In this case, both ions are coloured.
Such dye complexes will stain both nucleus and cytoplasm from a
single dye bath.
Romanowskystains are neutral dyes made from more complex
mixtures.
These are the commonest dyes used in haematology.
They are less common in histology but still very useful and include
Giemsa, Leishman and Wright’s stains.
NATURAL DYES
Natural dyes are simply dye substances extracted from natural sources.
They have largely been replaced by synthetic dyes, which are usually
more reliable, cheaper and can be supplied more readily.
Natural dyes still in use include haematoxylin, carmine, orcein and
litmus, although synthetic varieties are also available for some of these. 
Haematoxylin
Carmine
 
Synthetic or Artificial:
Nitroso (Naptholgreen)
Nitro (Picric acid)
Azo(Congo red, Orange G, Sudan III & IV)
Xanthine (Eosin, Rose Bengal)
Thiazole(Titan yellow)
Quonolin
Alcian blue