Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________

Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________
EXPERIMENT 1: BASIC TECHNIQUES IN MICROSCOPY

Objectives

i. To obtain accurate images

ii. To determine the depth of field
iii. To determine the field of view
iv. To calculate the actual magnification
v. To apply the use of oil immersion with high magnification (oil immersion lens)

Exercise 1.1: Images, Depth of Fields and Field of View of The Microscope

1.1.1 Images under the microscope

Apparatus
Compound light microscope

Materials
‘e’ prepared slide

Procedures
1. The ‘e’ prepared slide is observed using the 4x objective lens.
2. Observation under 4x objective lens is drawn.
3. The position of ‘e’ is determined.

Observation

‘e’ image

Actual magnification: ___40X_____

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________
Conclusion:

The letter ‘e’ image is _inverted_______ and __larger______.

1.1.2 The depth of field

Apparatus
Compaund light microscope

Materials
Cross threads prepared slide ( 3 colours eg : yellow , red and blue

Procedures

1. The position of the thread on the slide was observed with naked eyes.
The colour of thread was identified
a) At the top
b) In the middle
c) At the bottom

2. The cross threads was observe under the microscope using 4x and 10x
objective lens

Observation

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________

As for your reference, the above diagram shows the cross thread slide observed with naked
eyes and using 4x, 10x and 40x objective lens.
Follow the procedures given in the lab manual and state your observation in the table below.

Position of the naked eyes Identify the colour of thread

thread on the under the microscope using
slide 4x objective 10x objective 40x objective
lens. lens. lens.
at the top blue blue blue blue
in the middle red red red blue
at the bottom yellow yellow yellow yellow

Conclusion:

The ____bigger____ the magnification, the _bigger_______ the depth of field.

1.1.3 The field of view

Apparatus
Compaund light microscope

Materials
Transparent ruler ( 10 mm size )

Procedures

1. A transparent ruler is placed on the stage. The ruler scale is placed at the centre of
field of view.
2. The transparent ruler is observed using the 4x, 10x and 40x objective lenses. (The
amount of light is increased by adjusting the control knob to the maximum)
3. Obsevation is observed at 4x ovjective lens.
4. The diameter of field of view is determined at 4x, 10x and 40x objective lens
5. The formula is used to calculate the diameter of field of view

Diameter of field of view under

low magnification power = high magnification power
_______________________________ ______
Diameter of field of view under
high magnification power low magnification power

Observation

“ Transparent ruler “

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Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________

Actual magnification: 40x

Determine: (Show your calculation for question 2 & 3)

1. Diameter of field of view for the 4x objective lens is ______ mm or ______ µm.

2. Diameter of field of view for the 10x =______ mm = _______ µm.

3. The diameter of field of view for the 40x = ______mm = ______ µm.

Conclusion:

The __bigger______ the magnification, the __smaller______ the size of the field of view.

Exercise 1.2: Magnification

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________
Procedure

1. The actual magnification of a specimen was determined by

using the formula
Actual Magnification power = Magnification power objective lens x magnification power of ocular lens

2.Actual magnification power in table 1.1 was calculated

Observation

Table 1.1 Actual magnification of a specimen

Actual magnification
Magnification power of Magnification power of objective lens
ocular lens
4x 10x 40x 100x
10x 40x 100x 400x 1000x

Conclusion:

Using the 10x magnification power of __ocular______ lens, the greater the magnification
power of __objective______ lens will cause the actual magnification to increased________.

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________

Exercise 1.3: Oil Immersion Objective Lens

Apparatus
Compound light microscope

Materials
Immersion oil, lens tissue papers, prepared slide of bacteria, Methylated spiri

Procedures
1. The prepared slide was observed under the microscope using 100x objective lens

2. Observation is drawn

Observation:

“ Immersion objective lens”

Actual magnification: __1000x______

Conclusion:

Oil immersion techniques using higher magnification (__100x______ objective lens), where
oils ____reduce____ light refraction. The ______higher__ the resolution of the microscope,
the ____higher____ is the ability to distinguish details of the object.

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________

Questions:

Part A

For questions 1 to 7, choose the correct answer from the following list:
A Scanning objective lens (4x)
B Low-power objective lens (10x)
C High-power objective lens (40x)
D Oil immersion objective lens (100x)

1. Which is the shortest objective lens? ( ____A_____ )

2. Which objective lens should you use when you begin to focus a specimen?
( _____A____ )
3. Which objective lens should be in position before you store a microscope?
( ______A___ )
4. Which objective lens will deliver the highest amount of light? ( __A________ )
5. Which objective lens requires immersion oil to fill up the space between the specimen and
the lens? ( ______D____ )
6. Which objective lens will remain in focus when placed at the longest working distance
from the specimen? ( _______A___ )
7. When using an ocular lens with 10x magnification power, which objective lens should be
used to obtain the following actual magnification?
(a) 100 times of its diameter. ( ________B__ )
(b) 1000 times of its diameter. ( _______D___ )

Part B

1. Using 40x objective lens, determine the length and width of a cell from a piece of cork
tissue with approximately 20 cells in horizontal position and 10 cells in vertical position.

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Student’s name : ____NUR AIN BASHIRAH BINTI ZAHRUL HAKIM__________________
Class: ___B3T9_________
Lecturer’s name : _____PN WAN NORAFIFAH_________________

Figure 1.4 Cork tissue

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

We use lens cleaning fluid to clean the microscope lenses

3. 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)?

If the organism has moved out of the field of view to the right. I will move my slide to
left in order to keep observing the microorganism

4. How do you adjust the slide when the specimen is out of the field of view to the top?

I will adjust the slide to the bottom when the specimen is out of field of view to the top

Part C

1. Complete the following sentences.

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

b. Condenser and iris diaphragm are useful to coordinate the specimen

\References:

1 http://en.wikipedia.org/wiki/Bacillus_anthracis

2.. Campbell, N. A., Reece, J. B., Urry, L. A., Cain, M. L., Wassermen, S. A., Minorsky, P.
V. & Jackson, R.B. (2018). Biology. (11th Ed.). Pearson Benjamin Cummings. USA

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