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Bacterial Staining Techniques Guide

This document discusses staining procedures used in microbiology. There are two classes of ionizable dyes: basic dyes which are positively charged and bind to negatively charged molecules, and acid dyes which are negatively charged and bind to positively charged cell structures. Examples of staining techniques include simple staining using a single dye, differential staining to divide bacteria into groups, Gram staining which is the most common differential stain, and negative staining to demonstrate capsules or study morphology. Key steps in differential staining are application of primary stain, mordant, decolorizing agent, and counterstain.

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Liah Manlapig
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177 views2 pages

Bacterial Staining Techniques Guide

This document discusses staining procedures used in microbiology. There are two classes of ionizable dyes: basic dyes which are positively charged and bind to negatively charged molecules, and acid dyes which are negatively charged and bind to positively charged cell structures. Examples of staining techniques include simple staining using a single dye, differential staining to divide bacteria into groups, Gram staining which is the most common differential stain, and negative staining to demonstrate capsules or study morphology. Key steps in differential staining are application of primary stain, mordant, decolorizing agent, and counterstain.

Uploaded by

Liah Manlapig
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© © All Rights Reserved
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Staining Procedures • Inoculums size for staining: 10^5

cfu/ml
Classes of Ionizible Dyes:
• Examples: Gram staining and Acid
• 1. Basic Dyes Fast Bacilli staining
• Are commonly dyes used in Steps in Differential Staining:
bacteriology
1. Application of primary stain
• Cationic or have a positively charged
groups that bind to negatively 2. Application of Mordant
charged molecules like nucleic acids
3. Application of Decolorizing reagent
and proteins.

• Examples: methylene blue, basic 4. Application of secondary stain or


Counterstain
fuchsin, crystal violet, safranin and
malachite green. 3. diagnostic antibody or DNA probe Mediated
Staining
*positively charged groups (pentavalent nitrogen)

• 2. acid dyes -Directed specifically at identification of


an organism
• Anionic or possesses a negatively
charged groups that bind to -Used for the specific identification of selected
pathogens such as Chamydia, Bordetella, Legionella,
positively charged cell structures.
HSV, Varicella, CMV, adenovirus and respiratory virus
• Examples: eosin, rose Bengal and
• 4. Negative Staining
acid fuchsin

*negatively charged groups (carboxylic and phenolic) • Utilize to demonstrate the presence
of diffuse capsule surrounding some
Staining Techniques bacteria.

• 1. Simple Staining • Excellent technique to study


bacterial gas vacuoles and viral
• A single stain is used. morphology.
• It is directed towards coloring the • Also used to study cells sensitive for
forms and shapes present. heat fixing.
• Examples: malachite green, • Result: the bacteria appear as light
methylene blue, crystal violet, carbol colored bodies against a dark
fuchsin and safranin background.
• 2. Differential Staining • Example: India Ink or Nigrosin dye.
• It divides bacteria into separate
groups.
Gram Stain
• It is directed towards coloring
components of those elements • Most commonly used differential stain in the
present. clinical microbiology laborator

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