Riverside College Inc.

– Bacolod
College of Arts and Sciences
Microbiology and Parasitology

ACTIVITY NO. 1 – BASIC MICROSCOPY

Part I. Parts of the Microscope

Ocular Lens
(Eyepiece)
Head
(e

Objective
Lens

Diopter Adjustment Frame (Arm)

Nose Piece Arm

Stage Control
Mechanical Stage

Condenser Coarse Adjustment

Illumination

Brightness Adjustment
Fine Adjustment

Light Switch
Base
Figure 1. Different Parts of Microscope

Part II. Different Types of Microscope

Types Uses/s Source of Light/Illumination

is also known as the Compound The brightfield microscope uses

Light Microscope. With a compound visible light as a source of
Brightfield light microscope, we can examine illumination and the image appears
very small specimens as well as dark in the brighter background.
some of their fine details.

To study living cells without staining.
The ongoing different biological
Phase processes in the live cells can be
Contrast studied.
To study microbial motility.
Phase-contrast microscope is a
bright field-light microscope with the
addition of a special phase-contrast
objective (phase plate) and a phase
contrast condenser (annular stop).
When the light passes from one
substance to another substance
having a slightly different refractive
 Riverside College Inc. – Bacolod
College of Arts and Sciences
Microbiology and Parasitology
To observe endospores and
inclusion bodies that contain poly-
hydroxybutyrate,
polymetaphosphate, sulfur, or other
substances
is used to examine living
microorganisms that are invisible in
bright-field microscopy, do not stain
easily, or are distorted by staining.
Darkfield
is often used to image specific
features of small specimens such as
microbes.
To identify different bacterial
pathogens after staining them with
fluorochromes.
Fluorescence To distinguish live bacteria from
dead bacteria based on the color of
their fluorescence when they are
treated with a special mixture of
stains.
To study smaller objects such as
viruses or objects or molecules
having sizes smaller than 0.2 µm.
To study the details of the internal
Electron
structure of the cells.
To observe the ultrastructure of
microorganisms, large molecules,
biopsy samples, metals, and
crystals.
index or thickness, it will change the
phase. Phase-contrast
compromises between light intensity
and uniform phase by using a
circular ring (annulus) of
illumination. This annulus acted
similarly to a ring of pinholes, with
any particular direction around the
ring having the same phase, even
though the phase would vary
irregularly around the ring.
A dark-field microscope uses a
darkfield condenser instead of the
normal condenser. An opaque disk of
this condenser blocks the light that
would enter the objective lens directly.
The only light that is reflected or
refracted by the specimens enters the
objective lens and forms an image. The
field surrounding a specimen appears
dark and the specimen appears brightly
illuminated so the name dark-field
microscope.
A fluorescence microscope is much the
same as a conventional light
microscope but it uses light of higher
intensity as a light source instead of
visible light.
Commonly used light sources in
widefield fluorescence microscopy are
light-emitting diodes (LEDs), mercury or
xenon arc-lamps or tungsten-halogen
lamps. The most common light sources
used today are light-emitting diodes
(LEDs); their properties mean they are
usually superior in comparison to arc-
lamps and tungsten-halogen lamps,
which were more commonly used in the
past.
Electron microscopes use the electron
beam as an illumination source and
examine structures too small to be
resolved with light microscopes.
 Riverside College Inc. – Bacolod
College of Arts and Sciences
Microbiology and Parasitology
III. Questions
Part III. Questions
1. Explain the use of:

a. Low – power Objective

You can see more of the specimen with a low-power objective lens. Its main
purpose is to give the observer a wider range of the image so they may perceive
different characteristics of whatever specimen they are looking at. This objective
is useful for aligning the microscope. The power for the low objective is 10x.

b. High – power Objective

A high-powered objective lens, commonly referred to as a "high dry" lens, is the
best choice for observing fine details in a specimen sample. A high-power
objective lens and a 10x eyepiece magnify an object by a total of 400 times,
giving you a highly detailed view of the specimen in your slide.

c. Oil Immersion Objective

The oil immersion objective lens provides the most powerful magnification, with a
whopping magnification total of 1000x when combined with a 10x eyepiece. But
the refractive index of air and your glass slide are slightly different, so a special
immersion oil must be used to help bridge the gap. Without adding a drop of
immersion oil, the oil immersion objective lens will not function correctly, the
specimen will appear blurry, and you will not achieve an ideal magnification or
resolution. Oil immersion lenses are also available from some manufacturers in
lower magnifications, and provide higher resolution than their "high dry"
counterparts.

2. State what is meant by the “Focal length of the objective lenses”. How can you use this
information in focusing?
The distance from the center of the lens to the focal point is called the focal length. Focal
length is important because it determines the lens strength, which is an indication of how
much the lens enlarges the image. A lens with a shorter focal length will have a higher
lens strength and will enlarge the image more. Microscope objectives have short focal
lengths to greatly enlarge the images. Knowing this information will help me access the
microscope easily and effectively.

4. What are the corresponding ring colors of the objective lenses of the microscope?
Red band for a 4x scanning objective lens
Yellow band for a 10x low-power objective lens
Blue band is for a 40x high-power objective lens
White band for a 100x oil immersion lens.
 Riverside College Inc. – Bacolod
College of Arts and Sciences
Microbiology and Parasitology

Name of Student: Ruwela A. Parnoncillon

Program, Year & Section: BSN-2 Leininger
Date Submitted: August 25, 2023