Techniques in Microbiology: Staining Techniques

Staining of the clinical material or the bacteria from colonies on laboratory media
provide a direct visualization of the morphology of the organisms as well as their
reactions to the chemicals present in stains. This is an invaluable and easy-to-
use tool for establishing the identity of various microorganisms. Some of the
commonly-used staining techniques are:

staining affinity refers to the molecular attraction (or affinity) of a specific stain for a
particular type of cell component. an example would be the gram stain. the crystal violet
has an affinity for the molecules found in the cell wall of gram positive organisms, while
safranin has an affinity for the cell wall of gram negative organisms.

Gram's iodine stain is applied after the culture is stained with the primary stain. It acts as
a mordant, fixing the primary stain to the cell wall while lending no additional colour to
the cell (i.e. the mordant itself is not a stain).

The mordant is only able to fix the stain to Gram-positive bacteria because of the
characteristic thick, peptidoglycan coat that they possess. Because the mordant is not able
to fix the stain to Gram-negative bacteria (who's coat have a different composition), the
crystal violet stain will wash away from Gram-negative bacteria when the decolourizing
agent is added.

A counterstain is a stain with color contrasting to the principal stain, making the stained
structure more easily visible.