Course code : BWD11703

Section :2

Experiment 1 : Microscopy

Name/Group : Group 10

Member :1. Mohamad Said Bin Zambry CW220001

:2. Muhamad Khairil Shafiee Bin AW220172
Norhadi
:3. Afif Irfan Bin Muhd Nassri CW220030
:4. Nurul Nazifa Binti Hasenizam CW220004
:5. Nur Najwa Binti Mohd Rafie AW220179
:6. Nurul Mawaddah Binti Mat Nazri AW220224

Date of experiment : 26 October 2022

Instructor : Mrs. Fatariah Zakaria

NO. IMAGE NAME MATRIC NUMBER
1.

Mohamad Said Bin Zambry CW220001

2.

Muhamad Khairil Shafiee Bin AW220172

Norhadi

3.

Afif Irfan Bin Muhd Nassri CW220030

4.

Nurul Nazifa Binti Hasenizam CW220004

5.

Nur Najwa Binti Mohd Rafie

AW220179

6.

Nurul Mawaddah Binti Mat Nazri AW220224

 Work Distribution
NAME WORK

Nurul Nazifa Binti Hasenizam Introduction

Mohamad Said Bin Zambry Material and Methods

Nur Najwa Binti Mohd Rafie Observation

Nurul Mawaddah Binti Mat Nazri Discussion

Muhamad Khairil Shafiee Bin Norhadi Conclusion

Afif Irfan Bin Muhd Nassri Reference

Microscopy
Faculty of Applied Sciences and Technology, Universiti Tun Hussein Onn Malaysia, KM1 Jalan
Panchor, 86400 Pagoh, Muar Johor, Malaysia

Introduction

Many organisms bacteria and parts of organisms cells that biologist study are too small to be
seen with the human eye. There are several types of microscopes, but we will be mostly using
a compound microscope. This type of microscope uses visible light focused through two
lenses, the ocular, and the objective, to view a small specimen. Only cells that are thin enough
for light to pass through will be visible with a light microscope in a two-dimensional image.

Almost every microbe that exists is impossible to see with the naked eye, because it is
essentially invisible. To really see it, the use of a microscope is necessary, as well as knowing
how to use it. However, for the sake of this lab, a compound microscope is used for a
microscope that has two lenses between the eye and the slide. This allows for magnification
to occur and adequate lighting, which work together for easier viewing. For this laboratory,
to see certain microbes, the following "instruments" are used on the microscope is the base,
which includes the light source, the stage to hold the slide, the iris diaphragm allows to control
the light needed to see the specimen, the objective lens. There is also a coarse adjustment
knob that is used to focus low power objectives [4x and 10x], and the fine adjustment knob is
used to focus high power and oil immersion lenses. There is also magnification, found on the
rotating nosepiece, to help zoom in on the specimen and ranges from 4x to 100x

first, we ourselves know that there are many important components in the microscope that
we must know, for example Eyepiece ocular lens with or without Pointer. The part that is
looked through at the top of the compound microscope. Eyepieces typically have a
magnification between 5x & 30x. Then, Monocular or Binocular Head is Structural support
that holds & connects the eyepieces to the objective lenses. After then, Arm is Supports the
microscope head and attaches it to the base. Then, Nosepiece is Holds the objective lenses &
attaches them to the microscope head. This part rotates to change which objective lens is
active.

Then, Base Bottom base of the microscope that houses the illumination & supports the
compound microscope. Next, Objective lenses. There are usually 3-5 optical lens objectives
on a compound microscope each with different magnification levels. 4x, 10x, 40x, and 100x
are the most common magnifying powers used for the objectives. The total magnification of
a compound microscope is calculated by multiplying the objective lens magnification by the
eyepiece magnification level. So, a compound microscope with a 10x eyepiece magnification
looking through the 40x objective lens has a total magnification of 400x (10 x 40). After that,
Specimen, or slide. The object used to hold the specimen in place along with slide covers for
viewing. Most slides & slide covers are thin glass rectangles. Next, Stage or Platform. The
platform upon which the specimen or slide are placed. The height of the mechanical stage is
adjustable on most compound microscopes. Next, Stage clips or mechanical stage. Clips on
the stage that hold the slide in place on the mechanical stage. Aperture is disc or Iris
Diaphragm. Circular opening in the stage where the illumination from the base of the
compound microscope reaches the platform of the stage. Then, Abbe Condenser. This lens
condenses the light from the base illumination and focuses it onto the stage. This piece of the
compound microscope sits below the stage & typically acts as a structural support that
connects the stage to the arm or frame of the microscope. Next, coarse, and fine adjustment
controls is Adjusts the focus of the microscope. These knobs increase or decrease the level of
detail seen when looking at the slide or specimen through the eyepiece of the compound
microscope. Next, Stage height adjustment is to Adjusts the position of the mechanical stage
vertically & horizontally. It is important to adjust these knobs so that the objective lens is
never encountering the slide or specimen on the stage. After that, Mirror is Reflects light into
the base of the microscope. Earlier microscopes used mirrors that reflected light into the base
of the microscope instead of halogen bulbs as their source of illumination. Then, Illumination
is Light used to illuminate the slide or specimen from the base of the microscope. Low voltage
halogen bulbs are the most used source of illumination for compound microscopes and
bottom lens, or field diaphragm is knob used to adjust the amount of light that reaches the
specimen or slide from the base illumination.
2.1 The Compound Light Microscope

Method and apparatus

There are three materials used in this activity is light microscope, letter ‘e‘ cut out, microscope
slides and cover slips.

The activities began with obtaining the microscope slide with the letter “e“. Next, the slide
was placed on the stage and ensured that is locked in place with the slide holder. Then, the
condenser focusing knob was rotated to move the condenser to its highest position. After
that, the microscope was focused on any portion of the slide and then the condenser was
simply closed down and moved until a sharply focused view was obtained. Next, the
microscope was turned on by rotating the dimmer switch and the light intensity was adjusted
to a comfortable level. Then, the eyepieces were adjusted to get ocular distance and diopter.
After that, the eyes tube back was moved back or forth until saw one uniform field of view.

Next, the nosepiece was rotated to 10X magnification. Then, the coarse focus control was
rotated until the slide was close to the 10X objective as possible. The stage manipulators were
moved until a portion of the slide is directly under the objective. After adjusting the focus at
10X, the nosepiece was rotated to the next highest magnification. Next, the fine focus was
manipulated to obtain the sharpest image. Then, the slide was moved around until the letter
“e” was located in the view. The letter “e” was observed and recorded. After that, the object
was centered to the needed view and the nosepiece was rotated to the 40X objective position.
Lastly, the changes in the orientation of the letter “e” were observed and recorded.

Observation/Result

Microscope Observation/Result
Magnification Under Microscope

10x
 40x

Discussion

A microscope is an important tool in biology. The word microscope itself is from ancient Greek
that means to look at the tiny because the microscope is used to study things that are very
small and cannot be observed by the naked eye. The type of microscope that is commonly
used and we will be using in a lab is a compound light microscope. Light microscopes magnify
the image of specimen work using light that emit through two different lenses. the light will
be focused, and it is transmitted through the specimen. Lens that is closest with the specimen
is called the objective lens meanwhile the closest with the observer's eye is called the
eyepiece lens. When we observe the specimen using the compound light microscope it must
be placed on the glass slide.

From this experiment we were able to identify the parts of the image of the letter “e”. We
can describe the steps to view a slide on a compound light microscope. Also, we managed to
try and familiarize ourselves with the basic operations of a compound light microscope to
observe the specimen. There are three objective lenses with their marked magnification
power. The magnification power of an ocular lens of the microscope is 10x. As for the
objective lens, the lowest magnification power lens is 4x and the high magnification power
lens is 40x.

Based on this experiment, we noticed that the position of the image of the letter “e” under
the microscope is different from the position that we placed on the slide. When we are
watching the image over the microscope, the image we see is upside down and backwards.
We slightly turn the fine adjustment knob to focus the image of the letter “e” to focus and
obtain the sharpest image. We see less of the presence of the letter “e” that can be defined
as a magnified image of the letter “e”. Both magnification and constancy are necessary to get
a clear image of a very tiny thing. If you don't pay attention to the constancy of the microscope
but using a high magnification lens, you only get a magnified but fuzzy image.
2.2 Use of Oil Immersion
Method and Apparatus
There are three materials used in this activity is light microscope equipped with 100X, an
immersion lens, immersion oil and a pre-mounted bacteria slide. The sample used was
Bacillus Subtilis slide. The activities began with placed the prepared slide on the microscope
stage and ensure the light intensity adjusted to a comfortable level. By using the 10X
objective, the microscope was focus on an appropriate filed containing bacteria. The bacteria
slide was centred in the field of view by manipulating the lateral movement knobs. Then, the
nosepiece was rotated to the 40X objective and refocused with the fine focus. Next, the
nosepiece was rotated to obtain an intermediate position between 40X and 100X objectives.
After that, a small drop of immersion oil was placed on the centre of the viewing area of the
slide. Then, the nosepiece was rotated to the 100X, and the objective was rotated into the oil
by manipulating the lateral movement knobs. Lastly, the shapes of the bacteria were
determined and recorded.

Observation/Result

Microscope Observation/Result
Magnification Under Microscope

10x

40x
 100x
(under immersion oil)

Discussion

The objective lenses used for microscopes are parfocal which means that if one lens is focused
on the specimen on the slide, each of the other lenses is approximately in focus when
revolving the position. So, we only need to slightly turn to adjust the fine adjustment knob to
get the image in focus. To achieve parfocal for the microscope, you have to use the lenses in
sequence. For example, if you use the 4x lens and turn directly to 100x, parfocal will not be
achieved and it will require a lot of adjustment to sharpen the image. As the magnification
has increased, the iris diagram must have a wider opening to accommodate more light, so the
iris diaphragm must be opened deliberately until the ideal contrast and constancy are
achieved. As a result of obtaining an ideal contrast and constancy, the condenser has been
moved closer to the stage. To use the 100X magnification, immersion oil must be used.
Immersion oil is used to retain the light rays at the highest magnification because it has the
same refractive index as the glass, eventually, it also becomes a part of the optics in the
microscope. When using oil, the oil must be lightly touched with the objective lens. The
condenser should be in the upmost position and the iris diaphragm must be fully opened.

Conclusion

Microscopy is a technical field that involves the use of microscopes to view items and aspects
of objects that are not visible to the human eye. The light microscope is a very powerful tool
for understanding the structure and function of tissues and it is widely used in biomedical
science courses, research, and diagnostic laboratories. The tool that was used in this
experiment is the light microscope. The first part of this microscope that played a big part in
this experiment is the stage which is the platform that holds the slide containing the specimen
to be viewed (letter “e” and microbe). Next is the light that is located beneath the stage which
is used as a light bulb to view the specimen more precisely. Next, the body tube contains an
ocular lens and nosepiece with other objective lenses. The function of each lens is very based
on the magnification. The fine focus knob and the coarse were used to focus the image by
adjusting each of these parts accordingly. Adjustable diaphragm and condenser located
beneath the stage were used to regulate the amount of light that passes through and contains
two sets of lenses used to concentrate the light To make sure the specimen can be seen
through the ocular lens the light sources must pass through the diaphragm and condenser
before continuing up through the specimen.

Reference

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Www.labmate-Online.com. Retrieved November 4, 2022, from

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microtechniques/4/breaking-ne

Piggot, P. J. (2016). Bacillus Subtilis - an overview | ScienceDirect Topics. Sciencedirect.com.

https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-

biology/bacillus-subtilis

Shannon, R., & Ford, B. (2019). microscope | Types, Parts, History, Diagram, & Facts.

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