DIPLOMA FARMASI(A 8240)

LAB MANUAL

PHARMACEUTICAL MICROBIOLOGY(DFB 1012)

KOLEJ ANTARABANGSATEKNOLOGI DAN PROFESIONAL,

PERLIS MALAYSIA

PHARMACEUTICAL MICROBIOLOGY (DFB 1012)LAB 1

THE COMPOUND MICROSCOPE

INTRODUCTION

The most fundamental skill of microbiology is the use of the microscope. The definitionof a microscope is a device for magnifying objects that are too small to be seen with the nakedeye. The development, evolution, and the inventors involved in the microscope is quiteinteresting. Prior to 150, the magnifying glass was the best form of magnification and was notnecessarily that advanced. In 1500, Zacharias Janssen and Hans, his son, invented a microscopethat used two sets of lenses, a forerunner to the compound microscope. They used sunlight toilluminate the objects being studied. This earliest microscope magnified objects up to 100 timesthe objects actual size.

Todays microscopes often employ more than two sets. The largest and most accuratemicroscope is the electron microscope that uses electrons in a focused beam to illuminate objectsrather than light. The electrons pass through the specimen being viewed and are electricallyfocused. Normally the image is then broadcast to a television monitor or photographed. Ascanning electron microscope gives a three-dimensional image of the specimen. An electronmicroscope is capable of specimen magnification millions of times the actual size. This type ofmicroscopy has allowed the viewing of virus particles that are so small a few million could fit onthe head of a pin.

OBJECTIVES1) Learn proper care and handling techniques of the microscope.2) Identify and understand functions of selected microscope parts.3) Become proficient at focusing a microscope.4) Learn microscope terminology.5) Learn to use the microscope as a measuring tool.

ACA-OR-750-012-01KOLEJ ANTARABANGSA TEKNOLOGI & PROFESIONAL, PERLIS

MALAYSIAP/S 40 KM 5 JALAN KAKI BUKIT

01000 KANGAR, PERLIS

MATERIALS AND METHOD

1. Compound light microscope2. Prepared slides of 3 types of bacteria3. Lens paper4. Immersion oil

Procedure 1 - Using the Microscope

1. The part of the microscope was reviewed; each of the components was know the function andnames of the microscope.

2. Obtain a slide from the instructor.2. Place the slide on the stage of the microscope.3. Turn the revolving nosepiece so that the scope is on the 4x objective (low).4. Roll the stage up to its highest point.5. Move the stage until the specimen is in the middle of the stage.6. Focus on the specimen by rolling the stage down slowly using the coarse adjustment.

Procedure 2 - Examining Stained Bacteria

1. Obtain one slide of stained bacteria.2. Place the slide on the stage and follow proper procedure for focus, etc.3. Once the high dry objective (40x) has been reached and focus is maintained, apply one drop of

immersion oil to the slide and move to the oil-immersion objective (100x). Refocus ifnecessary and draw what is in the two fields on the report sheet.

OBSERVATION AND RESULT

1. Draw and label the compound light microscope2. Write how to proper care and handling techniques of the microscope.3. Give part of the microscope and their functions

REVIEW QUESTIONIt is often useful to know the exact amount of magnification of a particular object or organismviewed beneath the microscope. That is, how many times is the object enlarged. Totalmagnification can be calculated by using a simple formula:

Total Magnification = Ocular lens power x Objective lens power

Please complete the following to determine total magnifying power of each of the objectivelenses.

Table 1:

Magnification of eachObjective Lens Alone

Ocular lens Total Magnification

(Objective x Ocular)

Scanning Power

Low Power

High Power

4. Which objective focuses closest to the slide?

5. What controls the amount of light reaching the ocular lens?

6. Why should you always begin to use a microscope with the low-power objective?

7. Why should you never use coarse adjustment when focusing the high-power objectivelens?

Magnification of the ocular lens alone =

PHARMACEUTICAL MICROBIOLOGY (DFB 1012)LAB 2

CLASSIFICATION OF MICROORGANISM

INTRODUCTION

The term microbe is short for microorganism, which means small organism. To help peopleunderstand the different types of microbes, they are grouped or classified in various ways.Microbes are very diverse and represent all the great kingdoms of life.

Here are the major groups of microorganisms:

Viruses Bacteria Algae Fungi Protozoa

In this project you will study and experiment classification of bacteria by the shape and types ofthe bacteria colony.

OBJECTIVES:

1. To classify microorganisms into categories based on their characteristics.

MATERIALS

Prepared slides of the following bacteria: Bacillus Subtilis Escherichia coli Staphylococcus aureus Micrococcus luteus

MicroscopeOil immersion

ACA-OR-750-012-01KOLEJ ANTARABANGSA TEKNOLOGI & PROFESIONAL, PERLIS

MALAYSIAP/S 40 KM 5 JALAN KAKI BUKIT

01000 KANGAR, PERLIS

PROCEDURE

1. Using a microscope, look carefully at prepared slide of Bacillus Subtilis. Identify differentbacterial shape and arrangements.

2. Also, identify the size of the microorganisms that you are observe under microscope whenusing oil immersion.

3. Using oil immersion on microscopy 1000X lenses.4. Repeat the experiment with using another prepared slide.

OBSERVATION AND RESULTS

1. Make drawings of several of the bacteria from each of the four prepared slides andindicate their approximate size in micrometers.

Bacillus Subtilis Escherichia coli

Shape =

Types of microbe =

Shape =

Types of microbe =

Staphylococcus aureus Micrococcus aureus

Shape =

Types of microbe =

Shape =

Types of microbe =

REVIEW QUESTIONS

1. State three basic shapes of bacteria.

2. State five different arrangements of cocci.

3. State three different arrangements of bacilli.

4. State three different arrangements spiral form.