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7-Acid-Fast Stain

The document outlines a laboratory experiment for detecting Mycobacterium using an acid-fast stain, detailing the introduction, materials, and procedures involved. It also briefly describes other bacteria such as Corynebacterium diphtheriae, Clostridium tetani, Clostridium perfringens, and Bacillus anthracis, highlighting their characteristics. The experiment report includes sections for introduction, materials and methods, results, and bacterial morphology observation under a microscope.

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55 views11 pages

7-Acid-Fast Stain

The document outlines a laboratory experiment for detecting Mycobacterium using an acid-fast stain, detailing the introduction, materials, and procedures involved. It also briefly describes other bacteria such as Corynebacterium diphtheriae, Clostridium tetani, Clostridium perfringens, and Bacillus anthracis, highlighting their characteristics. The experiment report includes sections for introduction, materials and methods, results, and bacterial morphology observation under a microscope.

Uploaded by

jannatul80002
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPT, PDF, TXT or read online on Scribd

EXP 7 Mycobacterium detection

Acid-fast Stain
Introduction
• The cell wall of the genus Mycobacterium (causitive
pathogens of tuberculosis or lepra) is well endowed with
fatty or waxy substances (mycolic acid)
• The mycolic acid can resist decolorization with acid
alcohol.
• According to this property, acid-fast stain has been
devised to stain these bacteria.
• All of mycobacteria being stained are of red color, while
other bacteria and compounds in specimen are blue.
Materials
• Sputum from patients with tuberculosis
• Acid-fast Stain reagents:
1. Carbofuchsin
2. 3% acid alcohol (3 ml of concentrated hydrochloric
acid added into 97 ml of 95% alcohol)
3. methylene blue
• Marking pencil, Lens paper, bibulous paper, etc.
Procedures
1. Take 2 loop of the specimen, streak on a slide
2. Air dry and immobilization: pass the smear over flame
for 3 or 4 times (Note: don't overheat it)
3. Overlay carbofuchsin on the smears, heat the slide
above the flame, for about 5 min. (Note 2 items: not to
make the liquid boiling or dry).
4. Gently wash the slide with tap water and drain off the
water.
Procedures
5. Decolorize the smear with acid alcohol for about a
minute until the water draining from the slide become
colorless.
6. Gently wash with tap water.
7. Counterstain with methylene blue for 2 min.
8. Wash with tap water and gently blot up with bibulous
paper.
9. Examine the slide under oil-immersion objective.
Corynebacterium diphtheriae
→ Diphtheria
Albert stain
metachromatic
granules

• “English-letter” arrangement
• club-shaped: swelling at 1 end or both ends
• "barred" appearance: polyphosphate inclusions called
metachromatic granules (Babes-Ernst bodies)
• Clostridium tetani -
tetanus

• gram-positive
• terminal spores
• motile with peritrichous
flagella
Clostridium perfringens- gas gangrene

• Shape and structure


– Capsule
– Subterminal endospore
– Nonmotile
Bacillus anthracis
- anthrax
• Shape and structure
1. 1-3 ～ 5-10μm
2. Gram positive
3. Rod with square ends in
long chains (bamboo)
4. Central spore
5. Capsule (D-glutamate)
6. Nonmotile
Experiment Report
Part 1 Acid-fast Stain
1. Introduction
2. Materials and methods
Materials
Methods
3. Results
Draw a figure according the experiment result;
Intemperate it

Part 2 draw the morphology of bacteria only according to


what you have seen under the microscope

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