BACTERIAL STAINING

MARIA DYNAH T. PETILONA

RMT, EDD
 NEGATIVE
STAINING
Negative stain is also useful in determining bacterial
cell morphology.
Advantages:
Less shrinkage of the cells.
More accurate cell size dtermination.
Useful for spirochaetes
Leaving the orgnaisms transparent and visible in a
darkened field
 DIFFERENTIAL
STAINING
Differential staining reacts differently with
different kinds of bacteria and can be used to
distinguish them. Four types of reagents :
1. 1. Primary stain – initial stain applied to
bacterial smear.
2. 2. Mordant – increases the affinity of the
primary stain to the organisms.
3. 3. Decolorizer – washes off the primary stain.
4. 4. Secondary or counterstain – which gives
color to decolorized cell.
A bacterium is classified as Gram positive, if it
retains the color of the primary stain ( violet
or blue).
Gram positive cocci: Staphylococcus,
Streptococcus
Gram positive bacilli: Bacillus , Clostridium,
Corynebacterium, Lactobacillis, Listeria spp.
Gram negative if it decolorizes and takes up the color
of the counterstain (red or pink)

Gram negative cocci: Neisseria spp.

Gram negative bacilli: Escherichia, Pseudomonas,
Salmonella, Shigella, Haemophilus spp.
Acid fast organisms take up the color of the primary
stain (pink or red) while non acid fast take up the
counterstain (blue)