BIOLOGY

LAB REPORT
EXPERIMENT 1:
BASIC TECHNIQUES IN MICROSCOPY

NAME MOHAMAD AMMAR SHAFIQ BIN MOHAMAD SHAHRUL

AMMAR QAYYUM BIN MOHAMAD
DANISH QHALIFF BIN ADI WIRA
COLLEGE ID KMK2301132
KMK2301253
KMK2301270
PROGRAMME FIS-YTP
CLASS TUTORIAL FB 2
TEACHER/LECTURE NAME SIR ABD HALIM BIN ZAINAL
SUBMISSION DATE
Objective:

1. To observe images under the microscope (1.1)

2. To determine the depth of field (1.1)
3. To determine the field of view (1.1)
4. To calculate the actual magnification (1.2)
5. To observe cells from different tissues (1.3)
6. To learn how to observe fresh specimen (1.4)
7. To learn how to use the oil immersion objective lens (1.5)
8. To check the principles and procedures of using a microscope (1.6)

Introduction:

A laboratory is a controlled environment where experiments are conducted to gather data and
analyse phenomena. To ensure accurate results and maintain safety, proper techniques for
handling equipment is essential. This report focuses on key aspects of handling microscopes
correctly.

Exercise 1.1: Images, Depth of Fields and Field of View of the MMicroscope.

Apparatus:

Compound microscope

Materials:

‘e’ prepared slide

Cross threads prepared slide (3 colours)
Graph paper prepared slide 1 mm x 1 mm

Procedure

1. ‘e’ prepared slide was observed using a 4x objective lens.

2. The observation was drawn.
3. The position of ‘e’ was determined.
Observation:

The position of ‘e’ is inverted.

The depth of field

The depth of field refers to the thickness of the plane of focus. With a large depth of field, all of
the threads can be focused at the same time. With a smaller or narrower depth of field, only one
thread or a part of one thread can be focused, everything else will be out of focus. In order to
view the other threads, you must focus downward to view the ones underneath and upward view
the ones that are above.

Procedure

1. The cross threads were observed using a 4x objective lens.

2. The colour of the thread was noted. The location of the threads were identified as
a) Blue at the top
b) Yellow in the middle
c) Red at the bottom
3. The slide was removed and the position of the threads on the slide were observed with the
naked eye. The location of the threads were identified as
a) Blue at the top
b) Yellow in the middle
c) Red at the bottom

Observation:
The field of view

The simplest method of estimating linear dimension is to compare the size of the image to the
diameter of the field of view. You can make a rough estimate of the field diameter by focusing on
the millimetre scale of a transparent ruler using the lowest power objective. To calculate the field
of view, use this formula:

Procedure

The following exercise is done to determine the size of the field of view for each of the objective
lenses on the microscope.

1. A ‘graph paper’ prepared slide was placed on the stage.

2. The graph paper was observed using 4x, 10x and 40x objective lenses. (The amount of
light was increased by adjusting the control knob to the maximum)
3. A picture of the observation done using the objective lens was drawn.

Observation
Discussion

1. The diameter of field of view for 4x objective lens is 4.5 mm or 4000 um.
2. The size of field of view for the 10x and 40x objective lens is determined:
10x = 2.0mm = 2000um
40x = 0.4mm = 400um
3. If a piece of cork tissue with approximately 20 cells in horizontal position and 10 cells in
vertical position using a 40x objective lens. Determine the size of the cell.

Answer:
20 x 100 = 200
40x lens = 0.4 mm Field of view
Therefore, ( 0.4/200 ) = 0.002 mm size of the cell

Transparent ruler

Conclusion

In conclusion, the images under the microscope have been observed. Furthermore, the depth of
field has been determined. Finally, the field of view has been determined.
Exercise 1.2: Magnification

Procedure

1. The actual magnification when using a particular objective lens and ocular lens is calculated
using the following tables:

COMPOUND MICROSCOPE

Magnification
power of ocular Magnification power of objective lens
lens

4x 10x 40x 100x

5x 20x 50x 200x 500x

10x 40x 100x 400x 1000x

15x 60x 150x 600x 1500x

Conclusion

In conclusion, the actual magnification has been calculated.

Exercise 1.3: Cells

Apparatus

Compound microscope
Blade
Slide and cover slip

Material

Cork tissue
Stained prepared slides of animal and plant tissues

Procedures

Cork tissue

1. The cork tissue was sliced using a blade into very thin sections.
2. Placed a section on a slide.
3. 2-3 drops of water were added on the section and covered with a cover slip.
4. The cork tissue was observed under the microscope using 4x and 10x objective lens.
5. Observation was drawn and the magnification is stated

Stained prepared slides

1. Observed 9 prepared slides under 4x and 10x objective lens.

2. Observation was drawn.

Observation

Cork tissue

4x 10x
Stained prepared slides

Animal cell

Plant cell

Discussion

Cork tissue

Is your observation similar to what was observed by Robert Hooke? Describe your observation.

Answer : Yes, it is similar to what was observed by Robert Hooke. Boxlike cavities were
observed at the end of the cork.

Stained prepared slides

Is it easier to observe a stained tissue? What is the purpose of staining a specimen?

Answer : Yes, it is easier to observe a stained tissue. Staining a specimen helps to enhance
visualisation of the cell and other cell components under a microscope.

Conclusion

In conclusion, the cells from different tissues have been observed.

Exercise 1.4: Fresh Specimens

Apparatus

Compound microscope
Dropper
Concave slides and cover slips
Toothpick/ pin
Filter paper

Materials

Pond water from 2 different locations (containing microorganisms)

Procedures

1. Added a drop of pond water onto a clean slide.

2. Used a toothpick/pin to lower the cover slip gently so that air bubbles were not trapped.
3. Removed excess water using a filter paper to prevent the objective lens from touching the
water.
4. The different organisms present in the pond were observed.
5. Organisms that had been observed are drawn. The magnification is stated.

Observation

Conclusion

In conclusion, methods to observe fresh specimens have been learnt.

Exercise 1.6: Review Question

Instruction
Questions 1 to 7, choose the correct answer from the following :

A. Scanning objective lens (4x)

B. Low-power objective lens (10x)
C. High-power objective lens (Dry) (40x)
D. Oil immersion objective lenses (100x)

1. Which objective lens has the shortest length?

Answer : A

2. Which objective lens should be placed above the hole in the stage when you first begin to
focus the specimen or before you store the microscope? Assume the microscope you are
using has four objective lenses.

Answer : A

3. Which objective lens will deliver the highest amount of light based on the biggest
diameter of the lens?

Answer : D

4. The objective lenses should not become soaked with water or any other liquid, except for
a particular objective lens which will be immersed in oil to fill up the space between the
specimen and the objective lens. Name this objective lens?

Answer : D

5. Which objective lens will still remain in focus when placed at the longest distance from
the specimen?

Answer : A

6. If you use an ocular lens with 5x magnification power, which objective lens should be
used to obtain a magnification of 50 times of its diameter?

Answer : B
7.When using an ocular lens with 10x magnification power, which objective lens should be used
to obtain:

(a) A magnification of 100 times its diameter?

Answer : B

(b) A magnification of 1000 times of its diameter?

Answer : D

Answer the following questions and complete the sentences.

8.Based on laboratory practices, what do you use to clean the microscope lenses?

Answer: Lens cleaning fluid or cleaning mixture

9.A microscope is called a compound microscope when it consists of more than one set of lenses.

10.Condenser and iris diaphragm are useful to coordinate the angle of the beam of light focused
onto the specimen.

11. While observing a moving microorganism under a microscope, you found that the organism
has moved out of the field of view to the right. In order to keep observing the microorganism,
which direction do you move your slide (right/left)?

12.When the microorganism moves out of the field of view to the top, you can continue to
observe its movement.By moving the slide (away from you/towards you)