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BASIC DIAGNOSTIC & THERAPY SYSTEM

BY; NURHAFIZAH BT HATTA C 111 09 865

Practicing to do; - Preparation for Bacterial Smear - Hanging-Drop Preparation ( Examination of Living Bacterial Preparation) - Gram staining - Acid fast staining Observing normal flora of skin & mouth Disinfection & Sterilization Antimicrobial Susceptibilty Test

Aims;

- Identify the bacterial morphology and movement in a wet preparation (cell activity & fissions of cells; binary fission) - Differentiate between bacterial movement and Brownian motion

Bacteria are microorganisms with small size and with refraction index similar to the refraction index of water. This cause the bacteria are very hard to be seen alive. Thus, we observe them using the hanging-drop method to visualize the cells activity. However, we should know how to distinguish between the true motility or the Brownian movement (movement of the cells caused by collision with water).

TRUE MOTILITY

BROWNIAN MOVEMENT

Aims;

- To prepare smears from liquid and solid specimens for simple and gram staining Example specimens : Stool, etc.

1. 2. 3. 4.

5. 6.

Clean the slide with alcohol & pass through the flame, and label A, B and C. Turn the slide upside down (the label side is facing down). Put one or two drops of saline physiological solution on the object glass. Using a sterile loop, aseptically take a small amount of each bacterial culture A, B and C thus make a thin and even bacterial suspension on the slide. Air dry and heat fixed the smears on the slides. Slides have been prepared for the Gram staining.

Aims;

- To observe the bacterial shape (morphology) and aggregating pattern of bacteria. - To observe the Gram characteristics of bacteria.

Aims;

- To observe the shape and acid fast characteristic of bacteria


There

are two types of acid fast staining; a) Ziehl-Nielson method b) Tanzil / Kinyoun method

ZIEHL-NIELSON METHOD
Involve heat fixed process before staining

TANZIL METHOD
Do not involve heat fixed process before staining

Reagents used; i) Carbol fuchsin ii) Acid alcohol (ethanol 95% + HCl 3%) iii) Methylene blue

Reagents used; i) Kinyouns reagent ii) Acid alcohol (ethanol 95% + HCl 3%) iii) Methylene blue

Aim

- To observe the morphology and Gram characteristics of normal flora bacteria of human skin and mouth.
Example

of normal flora; Staphylococcus aureus

Prepare all the materials; for the medium agar, divide it into 4 quadrant

Observe the medium agar after 24 hours.

Aims;

- To compare the bacterial sensitivity towards the types of antibiotic with KirbyBauer method. ~ Do some measurements of clear zone around antibiotic disc (inhibition zone) then determine sensitivity characteristics from each antibiotic.

Practical

Work Guidelines, Basic Diagnostic & Therapy System 2009

Medical

Microbiology 24th Edition, McGraw Hill (LANGE)

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