BIO1204: GENERAL ZOOLOGY LAB

LABORATORY 1: MICROSCOPY AND OBSERVATION OF CELLS

Microscope

• A device used to view minute objects/organisms that are

otherwise invisible to the naked eye;

• Composed of lenses where small object can be magnified for

viewing.

Terminologies

• Virtual image – an image cannot be seen directly. It can only

be seen by a viewer looking through a lens.

• Real image – an image formed by the actual convergence of

light.

• Parfocal – construction of a microscope such that when an

image is focused with one objective in position, the other
objective can be rotated into place and the field will remain in
focus

• Monocular – a microscope with one eyepiece.t rays upon a

screen

• Transmitted illumination – light that passes up from the

condenser and though the specimen

• Vertical or reflected illumination – illumination of a specimen

from above; in microscopy it is used to examine opaque
specimens.

•Binocular – a microscope with two eyepieces.

• Field of view – the area of the specimen that can be seen

after it is magnified.

• Focus light rays at a specific place called the focal point.

• Distance between center of lens and focal point is the focal

length

• Strength of lens related to focal length.

• Short focal length → more magnification.

Focal length and Focal Point

Parts and Function

 BIO1204: GENERAL ZOOLOGY LAB

LABORATORY 1: MICROSCOPY AND OBSERVATION OF CELLS

Types of Microscopes

Bright Field
Images formed under the LPO, HPO, OIO -Cannot resolve organisms less than 0.2 um such as viruses

-Light Absorption is the main means of providing image

contrast.

-operates through the reflection of light from the objects to our

eyes.

Darkfield

- the objects are illuminated only from the sides and light is
reflected off the object being observed.

- useful in observing living organisms in a suspension.

 BIO1204: GENERAL ZOOLOGY LAB

LABORATORY 1: MICROSCOPY AND OBSERVATION OF CELLS

Fluorescence Scanning EM X10,000 Specimen can be

view in screen; 3-
- used to detect chemical bonding between certain substances. dimentional view;
- uses a special dye. surface
examination;
Stereoscope limited resolution
than transmission.
- It is used for dissection to get a better look at the larger
specimen. You cannot see individual cells because it has a low
magnification. Magnification

TEM) Transmission Electron microscope • magnitude of amplification of the image of the specimen.

Scanning Electron Microscope • It is also how much an image is enlarged under a microscope.

SEM Images Total Magnification

It is the number of the times the specimen is magnified

Types Magnification Characteristics

Brightfield X1000 Cannot resolve

organisms less
than 0.2 um such Resolution
as viruses.
Darkfield X1000 Back ground is • ability of the microscope to separate two or more clustered
dark & unstained objects so that each can be seen distinctly.
organisms can be
seen; difficulty in • is the amount of detail you can see in an image.
operation.
Phase Contrast X1000 Can observe • Depends on:
dense structures
»Wavelength of light λ = R
in living
prokaryotic & »Numerical Aperture (NA) - characteristic of the lens system.
eukaryotic
organisms. Micrometry
Fluorescence X1000 Fluorescent dye
attached to • A technique used to measure the dimensions of the
organisms; microorganisms.
training required
in preparation. • Uses two kinds of micrometer.
Transmission EM X200,000 Specimen can be
view in screen; Ocular micrometer - measures the dimensions of the
image is 2- specimen.
dimensional;
allows cellular Stage micrometer - calibrates the ocular micrometer.
ultrastructure
examination;
specimen is non-
living
 BIO1204: GENERAL ZOOLOGY LAB

LABORATORY 1: MICROSCOPY AND OBSERVATION OF CELLS

8. Looking through the eyepiece, VERY SLOWLY the

coarse adjustment knob until the specimen comes
into focus.
9. To switch to the LPO, look at the microscope from the
side. CAREFULLY revolve the nosepiece until the
high-power objective lens clicks into place.
10. Looking through the eyepiece, turn the fine
adjustment knob until the specimen comes into focus.

Care and Handling

• When you pick up the microscope and walk with it, grab the
arm with one hand and place your other hand on the bottom of
the base DON'T SWING THE MICROSCOPE

• Never touch the lenses with your fingers. Your body produces
an oil that smudges the glass. This oil can even etch the glass
if left on too long. Use only lens paper to clean the glass.

TOILET PAPER, KLEENEX, AND PAPER TOWELS HAVE

FIBERS THAT CAN SCRATCH THE LENSES.

Using the Microscope

1. To carry the microscope grasp the microscopes arm

with one hand. Place your other hand under the base.
2. Place the microscope on a table with the arm toward
you.
3. Turn the course adjustment knob to raise the body
tube.
4. Rotate the nosepiece until the low power objective
lens clicks into place.
5. Adjust the diaphragm. While looking through the
eyepiece.
6. Place a slide on the stage Center the specimen over
the opening on the stage. Use the stage clips to hold
the slide in place.
7. Look at the stage from the side. Carefully turn the
course adjustment knob to lower the body tube.