Basic Staining

Techniques in
Histology
 THE
MEMBERS
Batallones, Lemuel Rico, Samuel Santillan, Shan
Potente, Alexis Paul Saril, Alf Gabriel Tutaan, Karl Owen
Basod, Mart Jones

Source: Centers for Disease Control and Prevention

 What is
staining?
• The basic staining in
microbiology involves using
dyes to color cells or structures,
making them more visible under
a microscope.

Source: Centers for Disease Control and Prevention

What are its It is important to microbiology and other
significance? biological sciences for several reasons:

1 arrangement more clearly through enhanced visibility.

To allow researchers to observe their morphology, structure, and

Identification and Classification: Staining methods aid in identifying and

2 classifying microorganisms based on differences in their cell wall
composition.

3 Staining techniques play a vital role in clinical microbiology for

diagnosing infections. Different stains can help identify specific
pathogens in patient samples

Source: Centers for Disease Control and Prevention

Keys terms
Dye stains materials or less selectively and behave like acidic or basic.

Anionic- chemical substances that possess a negative charge ion

Cationic- chemical substances that possess a postitive charge ion.

Basophilic- cell components with a net negative charge that have

an affinity for basic dyes.

Acidophilic- chemical substances that possess a negative charge ion

Source: Centers for Disease Control and Prevention
The most common dyes:

Hematoxylin
Basic Dye
Dark blue or dark purple in color
Positively charged and stains nucleic acid and
Ribosomes

Source: Centers for Disease Control and Prevention

 Eosin
Acidic Dye
Pink in color
Negatively charged and stains cytoplsms and
proteins, such as collagen and elastin pink.
The Staining
Techniques

Source: Centers for Disease Control and Prevention

 Hematoxylin and
Eosin (H&E)
• The most common staining
technique in histopathology

• uses a combination of two dyes, used for demonstration of

nucleus and cytoplasmic inclusions in clinical specimens.

• Best used for human samples.

Source: Centers for Disease Control and Prevention

 Periodic acid-schiff
• widely used technique for the
demonstration of carbohydrates and
carbohydrate rich compounds in tissues.

• Stains in purple or magenta

Source: Centers for Disease Control and Prevention

Endospore Staining • used to determine the presence of
endospores in the bacteria sample

Source: Centers for Disease Control and Prevention

 Gram Staining
• -it is used to help detect the bacteria -
used to determine gram-positive and
gram-negative bacteria

• Often the first test performed, gram

staining involves the use of crystal
violet or methylene blue as the primary
color
Gram Staining • Often the first test performed, gram
staining involves the use of crystal
violet or methylene blue as the primary

Procedure: color

• The first step in gram staining is the use of crystal violet dye for the
slide's initial staining.
• The next step, also known as fixing the dye, involves using iodine to
form crystal violet- iodine complex to prevent easy removal of dye.
• Subsequently, a decolorizer, often solvent of ethanol and acetone, is
used to remove the dye.
Simple Staining
• involving only one stain/solution

• stains commonly used in staining are

crystal violet, safranin, and methylene
blue.
 Simple Staining procedure
1. Place the prepared fixed smear slides on the stain rack over the sink.
2. Let the stain sit for 1-5 minutes.
3. Using the clothespin, grab the long end of the slide, tilt the slide over the sink and
rinse the stain off with a stream of water from the wash bottle.
4. Touch the edge of the slide to a paper towel to remove excess water. You can now let
it air dry. Alternately you can dry it with blotting paper by placing it in the blotting paper
book and pressing lightly. While this method is quicker, you can also blot off a poorly
adhered smear.
5. View your slide under oil immersion and record your observations
REFERENCES
Smith, J. (2023). Staining Techniques in Histology: Hematoxylin and Eosin (H&E)
Staining. Journal of Cellular Biology, 15(3), 123-137.

Smith, A. C. (2018). Periodic Acid-Schiff (PAS) Staining: Principles and Applications

in Histology. Journal of Histotechnology, 42(3), 150-165.

Brown, A. B. (2010). Historical Development and Principles of the Gram Stain.

Microbiology Perspectives, 23(2), 45-56.

Johnson, M. P. (2015). Silver Staining for Protein Visualization in Polyacrylamide

Gels. Biochemical Techniques, 32(4), 210-225.