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Clinical Bacteriology Staining Guide

Staining techniques are used to differentiate bacteria and include simple staining with one dye and differential staining using multiple dyes. Gram staining, developed in 1884, uses crystal violet, iodine, alcohol or acetone, and safranin to stain the cell walls of gram-positive bacteria purple and gram-negative bacteria red. Acid-fast staining uses carbolfuchsin and identifies bacteria with mycolic acid in their cell walls such as Mycobacterium.

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118 views3 pages

Clinical Bacteriology Staining Guide

Staining techniques are used to differentiate bacteria and include simple staining with one dye and differential staining using multiple dyes. Gram staining, developed in 1884, uses crystal violet, iodine, alcohol or acetone, and safranin to stain the cell walls of gram-positive bacteria purple and gram-negative bacteria red. Acid-fast staining uses carbolfuchsin and identifies bacteria with mycolic acid in their cell walls such as Mycobacterium.

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lorraine del rosario
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We take content rights seriously. If you suspect this is your content, claim it here.
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MLS 044 - Clinical Bacteriology (LABORATORY) Staining Techniques

Staining Techniques
 Simple Staining
 application of a single staining solution
 basic dyes have greater affinity for nuclei; acid dyes have greater
affinity for cytoplasm

 Differential Staining
 Application of 2 or more stains; aids in differentiation of one group
to another
 Makes use of a primary stain, a mordant, decolorizer and
counterstain.

Gram Staining
 Developed by Danish bacteriologist Hans Christian Gram in 1884
 Stains the teichoic acid present in cell walls of Gram positive bacteria
 Not applicable to organisms that exists almost exclusively within host
cells, those that lack cell wall, and those of insufficient dimension to be
resolved by light microscopy

Reagents
Primary Stain Crystal Violet
Mordant Gram’s Iodine
Decolorizer Alcohol and/ or
Acetone
Counterstain Safranin

Staining Techniques KYD Staining Techniques


MLS 044 - Clinical Bacteriology (LABORATORY) Staining Techniques
bacilli
Gram positive bacilli Gram negative bacilli
Aerobes: Aerobes:
 Listeria  Acinetobacter
 Bacillus  Aeromonas
 Corynebacterium  Alcaligenes
 Mycobacterium  Bordetella
 Erysipelothrix  Brucella
 Nocardia  Enterobacteriaceae
 Legionella
 Pasteurella
 Psedomonas
 Vibrio
Gram Positive Gram Negative Anaerobes: Anaerobes:
 Clostridium  Fusobacterium
cocci  Lactobacillus  Bacteroides
Gram positive cocci Gram negative cocci  Actinomyces
Aerobes: Aerobes:  Propionibacterium
 Staphylococcus  Neisseria
 Streptococcus  Branhamella Acid-Fast Staining
 Micrococcus
 Stains the mycolic acid in the cell wall of bacteria
Anaerobes: Anaerobes:  Acid fast organism: Mycobacterium, Nocardia, Cryptosporidium,
 Peptococcus  Veillonella Isospora, Cyclospora.
 Peptostreptococcus
 Sarcina

Staining Techniques KYD Staining Techniques


MLS 044 - Clinical Bacteriology (LABORATORY) Staining Techniques
Ziehl-Nelseen Kinyoun
Method Method

Primary Stain Carbolfuchsin Carbolfuchsin

Mordant Steam Tergitol

Decolorizer HCl, alcohol HCl, alcohol

Counterstain Methylene Methylene


Blue Blue
Acid-Fast Positive Bacilli Acid-Fast Negative Bacilli

Staining Techniques KYD Staining Techniques

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