LABORATORY REPORT

BIOLOGY FOR PRE SCIENCE

BIOLOGY 083

TITLE OF EXPERIMENT : HANDLING AND OPERATING THE

MICROSCOPE AND LABORATORY SAFETY
DATE OF EXPERIMENT :
LECTURER’S NAME : MISS NUR HANIS BINTI MOHD
YASIM
NAME : NURFATIN NADHIRAH NAJAA’ BT
ADZHAR
CLASS : AS008/1A
MATRIC NUMBER : 2022474652
PRACTICAL 1 – HANDLING AND OPERATING THE MICROSCOPE AND
LABORATORY SAFETY
OBJECTIVES:
i. To label and describe the function of basic parts of the compound light microscope.
ii. To list in proper order the steps involved in microscope operating and maintenance.
iii. To calculate the diameter of the field of view and the total magnification for low and
higher power lens system.
APPARATUS: Light microscope, slide and coverslips, scissors, filter paper, tissue paper.
MATERIALS: Old newspaper, toothpick, distilled water, staining reagents.

INTRODUCTION:
Organisms can be very small ranging from millimeter (mm) down to even micrometer (µm).
This group is known as microorganism and a microscope is used to view their microscopic
features. Different instruments ranging from the hand lens to the electron microscope are
effective as magnifying devices. The brightfield microscope perhaps, the most commonly used
type of microscope in student laboratories, is a compound microscope that produce a dark
image on a bright background.

A scientist first developed a microscope known as Antony van Leeuwenhoek. He is known

as the “father of Biology” and also the first microbiologist because of his development of the
microscope and also involve on microbiology research by describing single-celled organisms.

Light microscope uses light rays that are magnified and focused by different types of lenses.
The binocular dissecting microscope is specially designed to study an object in three
dimensions under a relatively low levels of magnification. The compound light microscope is
commonly used for examining small or thinly-sliced sections of objects under higher
magnifications. The source of illumination comes from the bottom part of the microscope and
the light rays passed through the objects reflecting the complete feature of the specimen.
Usually to improve contrast, stains or dyes that bind to cellular structures and absorb the light
rays used.
A. PARTS OF THE MICROSCOPE

1. Eyepiece (ocular lens): Topmost series of lenses through which an object is viewed.
The ocular lenses typically magnify images 10 times (10x).
2. Body tube: Holds nosepiece at one end and eyepiece at the other end. The function of
this part is to conduct light rays.
3. Arm: Supporting the upper parts and provides carrying handle.
4. Nosepiece: Revolving device that holds objective lenses.
5. Objectives (objective lenses): Individual objectives are used to examine the
specimen’s feature under different magnifying power.
a) Scanning power objective: Holds 4x lens used to scan the whole slide.
b) Low-power objective: Holds 10x lens to view an object in greater detail.
c) High-power objective: Holds 40x lens used to view the object in even greater
detail.
d) Oil immersion objective: Holds the 100x lens and is used in conjunction with
immersion oil to view objects with greatest magnification allowed by the light
microscope.
6. Coarse-adjustment knob: Knob is used to bring the object or the stage into
approximate focus used only with low-power objective.
7. Fine-adjustment knob: Knob is used to bring the object into final focus.
8. Diaphragm or diaphragm control lever: Control the amount of illumination used to
view the object.
9. Light source: An attached lamp that produces a direct beam of light, passed through
the object and reflects their microscopic feature.
10. Base: The flat surface at the bottom of the microscope for better balance on the table.
11. Stage: Holds and support microscope slides.
12. Stage clips: Holds slide in a fixed place and prevents from any sliding movements.
13. Mechanical stage (optional): A moveable stage that fits with the size of the of the
slide and helps to hold the slide during the movement of the stage.
14. Mechanical stage control knob (optional): Two knobs that are usually located below
the stage. Each of them controls the stage to move forwards/in reverse/right/left.
B. SLIDE PREPARATION (LETTER ‘K’ SLIDE) AND STAINING
1. A drop of water was placed on the middle of a slide.
2. A small letter of K from a piece of newspaper were cut and placed on top of the water
for a few seconds.
3. One edge of the coverslip was carefully placed on the slide and lowered over the drop
to prevent the formation of air bubbles. This can be done with the assistance of a pin,
needle or toothpick.
4. This technique was practiced until no more bubbles being trapped under the slide cover.
5. 1-2 drops of staining were dropped at the edge of one end of the coverslip once the slide
with the letter ‘k’ were ready.
6. A piece of filter/tissue were placed at the other end of the slide. Staining would travel
across through the coverslip by capillary action and absorbed by the tissue paper.

C. PROCEDURE OF MICROSCOPE HANDLING

1. A microscope was placed on the table.
2. The socket was plugged into the power source.
3. The light was switched on and adjusted to appropriate brightness.
4. The condenser aperture diaphragm was fully opened.
5. A slide with the letter ‘k’ were placed on the mechanical stage by pulling out the slide
holder. The letter ‘k’ was situated right under the objective.
6. The specimen was found and focused first under the low power. Then, the slide was
positioned so the letter ‘k’ was exactly centered in the field of vision.
7. The lowest powered objective (4x or 10x) were placed in position.
8. The stage was moved up using the course adjustment so the letter ‘k’ came into focus.
9. The fine adjustment was adjusted to get a sharper focus.
10. To reduces eyestrain, both eyes were opened while looking through the eyepiece.
11. By revolving the nosepiece, the power objective was changed to higher one(40x).
12. The letter ‘k’ was focused using the fine focus only, until a sharp image acquired.
13. Magnification of the specimen were described. Eg: (10x) x (40x) = 400x magnification.
14. The microscope was returned to the lowest power objective and the slides were
removed from the stage, once finished.
D. OBSERVING POND WATER SAMPLE
1. A drop of pond water was placed on a glass slide using a dropper/pipette Pasteur.
2. A coverslip was placed over the specimen and examined under the low-power and high-
power magnification.
3. Microorganism observed and identified were listed.

RESULTS/OBSERVATIONS
A. LABEL THE MICROSCOPE

9.Eyepiece

1.bodytube

2.Nosepiece
10. Arm
3. Low power objective

4. High power objective

11. Stage
5.Oil-immerson objective
12. Course-
6.Stage clips
adjustment knob

7.Diaphragm 13. Fine-adjustment

knob
8.Light
source
14. Base
B. QUESTIONS
1) Describe the image under low power lens briefly
a) Does the image appear upside down compared to the object on the slide?

Ans: Yes.

b) Does the image appear laterally inverted compared to the object on the slide?

Ans: Yes.

c)Does the movement of the slide corresponds with the image movement under the
microscope?

Ans: Yes.

2) Calculate the total magnification of image observed below:

3) List of microorganisms identified: