Republic of the Philippines

Department of Science and Technology

PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

INTEGRATED SCIENCE 2 – 1st Quarter

AA#2: BASIC MICROSCOPY

"Micro" refers to tiny, "scope" refers to view or look at. Microscopes are tools used to enlarge images of
small objects so as they can be studied. Historians credit the invention of the compound microscope to the
Dutch spectacle maker, Zacharias Janssen, around the year 1590.The compound light microscope is an
instrument containing two lenses, which magnifies, and a variety of knobs to resolve (focus) the picture.
Because it uses more than one lens, it is sometimes called the compound microscope in addition to being
referred to as being a light microscope. In this lab, we will learn about the proper use and handling of the
microscope.

OBJECTIVES

a) Demonstrate the proper procedures used in correctly using the compound light microscope.
b) Prepare and use a wet mount.
c) Determine the total magnification of the microscope.
d) Explain how to properly handle the microscope.
e) Describe changes in the field of view and available light when going from low to high power using the
compound light microscope
f) Explain the proper procedure for focusing under low and high power using the compound light microscope.

MATERIALS

Microscope, glass slide, prepared slides

Figure 1. Parts of the microscope

PARTS OF THE MICROSCOPE

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph
 Republic of the Philippines
Department of Science and Technology
PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

1. Arm and base.

All other parts of the microscope are attached to the arm or base.

2. Mechanical stage: The platform on which the microscope slide rests and the clamping device that
secures the slide.

3. Mechanical stage control knobs (x-y control knobs): These knobs, under the stage, move the
stage front to back and the slide from side to side.

4. Lamp (illuminator).

The lamp on/off switch is located on the base. The light intensity can be adjusted with this on/off
switch. The lamp should be adjusted to a medium level at the start of viewing. The lamp or illuminator
usually has a blue filter that rests on the light housing or under the condenser.

5. Iris diaphragm: adjusts the amount of light reaching the specimen. It is adjusted with a thin, black
lever under the stage. It has a dramatic effect on the contrast observed in the specimen and may need to
be adjusted frequently.

6. Lens systems: There are three lens systems: the eyepieces (ocular), the objectives (four), and the
condenser.

a. Eyepieces (ocular): Magnification of 10X. The eyepieces are held rather loosely in the
eyepiece tubes. Never remove the eyepieces from the eyepiece tubes. A rubber eye shield
should be on the top of each eyepiece. The width between the eyepieces should be moved until a
full circle (the viewing field) is visible with both eyes simultaneously.

b. Objectives: There are four objectives: 4X (red), 10X (yellow), 40X (blue), and 100X (white,
oil immersion objective). The objectives are attached to a rotating nosepiece (nose turret).

The total magnification is calculated by multiplying the

ocular magnification and the magnification of the objective in use.

c. Condenser: The condenser is located directly beneath the stage. It gathers and conducts
the light to the specimen. Although it can be raised and lowered with the condenser adjustment
knob, the condenser should remain at its highest position.

7. Focusing knobs: located on both sides of the microscope.

The larger, inner coarse adjustment knob moves the stage up and down much faster and farther
than the smaller, outer fine adjustment knob. The coarse adjustment knob is used ONLY with the low
power (4X, 10X) objectives. When focusing under the 40X or 100X objective, ONLY use the fine
adjustment, never the coarse adjustment.

8. Nosepiece: Holds the objective lenses

USE OF THE MICROSCOPE

NOTE: When lifting the microscope, always grasp the arm of the microscope with the stronger hand and
support the base with the weaker hand. Never tilt the microscope or run it when carrying it.

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph
 Republic of the Philippines
Department of Science and Technology
PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

1. Place the microscope in an upright position with the arm directed towards you. Make sure that it is at
least two inches from the table.

2. Looking from the side, turn the revolving nosepiece clockwise until the LPO is in place.

3. Look through the eyepiece with one eye while keeping the other eye open.

4. Place the prepared slide on the stage in such a way that the part to be observed is centered over the
hole in the stage.

5. By tuning the coarse adjustment knob, rack the LPO or the stage all the way down. Do this while looking
from the side to prevent the unlikely event of breaking a slide.

6. Look into the eyepiece with one eye and slowly rack the LPO upward using the coarse adjustment knob
until the specimen is visible. Obtain a clear focus by slowly turning the coarse adjustment knob clockwise or
counterclockwise.

7. For greater magnification, turn the nosepiece until the HPO is in place. Take care to avoid breaking the
slide in place. Use the fine adjustment knob to increase resolution.

8. After using the microscope, makes sure to rack the body tube away from the stage using the coarse
adjustment knob. Remove the slide and wipe the stage with a clean tissue when wet.

9. Position the clips and the mirror. Place the objective with the lowest objective in position.

10. Make sure to close the diaphragm and rack the body tube closer to the stage.

LAB WORK

1. Obtain a prepared slide with the cut-out letter “e”. Place the slide in the stage in such a way that the
mounted letter “e” is its normal reading position. Hold the slide using the clips.

Draw the resulting image under the LPO. Describe the appearance of the letter as seen through the LPO.
While looking through the eyepiece, observe in what direction the image moves if the slide is slowly shifted:

a. to the right
b. to the left
c. upwards
d. downward

Return the letter “e” to the center of the field. Without moving the coarse adjustment knob, shift the
objective from LPO to HPO. Sharpen the focus using the fine adjustment knob. Draw the resulting image of
the letter “e” under HPO.

2. Draw the four (4) sample slides as viewed under the LPO and the HPO.

Name of Members: __________________________ , __________________________ Date: ________

__________________________ , __________________________ , __________________________

INTEGRATED SCIENCE 2 – 1st Quarter

AA#2: BASIC MICROSCOPY

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph
 Republic of the Philippines
Department of Science and Technology
PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

1. Microscopic observation of the letter “e”

NAME OF 1st MEMBER: ___________________________

Observation under LPO Observation under HPO

Magnification: ________x Magnification: ________x

A. While looking through the eyepiece, in what direction does the image of the letter moves while the slide
is slowly shifted:

a. to the right? _______________________________________________________

b. to the left? ________________________________________________________
c. upward? __________________________________________________________
d. downward? ________________________________________________________

B. Is there a change in the level of brightness of the field of view when the objective is shifted from LPO to
HPO? Describe the change.
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

C. To ensure easier focusing, what should be done first before HPO is put into position?
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

2. Draw the specimens from the prepared slides given.

a. Name of Sample Slide #1: ________________________

NAME OF 2nd MEMBER #2: ___________________________

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph
 Republic of the Philippines
Department of Science and Technology
PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

Observation under LPO Observation under HPO

Magnification: ________x Magnification: ________x

General observations:
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

b. Name of Sample Slide #2: ________________________

NAME OF 3rd MEMBER: ___________________________

Observation under LPO Observation under HPO

Magnification: ________x Magnification: ________x

General observations:
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

c. Name of Sample Slide #3: ________________________

NAME OF 4th MEMBER: ___________________________

Observation under LPO Observation under HPO

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph
 Republic of the Philippines
Department of Science and Technology
PHILIPPINE SCIENCE HIGH SCHOOL - CENTRAL LUZON CAMPUS
ISO 9001:2015 Certified
Certificate No. SCP000394Q

Magnification: ________x Magnification: ________x

General observations:
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

d. Name of Sample Slide #4: ________________________

NAME OF 5th MEMBER: ___________________________

Observation under LPO Observation under HPO

Magnification: ________x Magnification: ________x

General observations:
_____________________________________________________________________________________
_____________________________________________________________________________________
_____________________________________________________________________________________

Postal Address: Lily Hill St. Clark Freeport Zone, Tel. Nos.: (045) 499-0136
Pampanga, Philippines (045) 499-5597
Website: https://clc.pshs.edu.ph