Answer the following questions to enhance more your understanding in the field of microscopy.

Cite your reference/s in APA format.

1. What is a microscope?
Microscope is an instrument that displays amplified images of minute objects. It enables
us to examine details that are too small to be seen by the naked eye.

Ford & Shannon (2022) states that microscope came from the Latin word
“microscopium,” which is derived from Greek words: “micros,” meaning small, and “skopein,”
meaning “to look at.”

2. In a tabular form, give the description and function of the following parts of a
compound microscope:

A. MECHANICAL PARTS

MECHANICAL PARTS DESCRIPTION FUNCTION

1 Draw tube According to biology online, the draw It is used to control tube
tube is the smaller of the two tubes on length.
a monocular microscope.

2 Body tube Body tube comprises a hollow and It assures continuous

tubular structure. alignment of the
objective of the optics.

3 Revolving It is a rotating turret that is connected It is used to select

nosepiece to the body’s lower end. different objective
lenses.

4 Coarse adjustment Coarse adjustment knob is a large Coarse adjustment knob

knob knob that is used for moving the body allows the specimen to
tube closer or farther to the specimen. be examined under exact
focus.

5 Fine adjustment Fine adjustment knob is a knob smaller Fine adjustment knob is
knob than a coarse adjustment knob. used for sharp and fine
focus.

6 Handle/arm A curved structure that connects the It provides support to

body tube to the base of the the microscope and it is
microscope. also when carrying a
microscope.
7 Stage Stage is a flat and rectangular plate It is where the specimen
where the specimen is placed. is located upon
observation.

8 Clips Clips are two metals located at the It holds the slide in
stage. position. It prevents the
slide from moving while
viewing it under the
microscope.

B. ILLUMINATTING PARTS

MECHANICAL PARTS DESCRIPTION FUNCTION

1 Ocular/ eyepiece The eyepiece (or ocular lens) is the lens part Ocular/eyepiece
at the top of a microscope that the viewer magnifies the image
looks through. The standard eyepiece has a produced by the
magnification of 10x. You may exchange with microscope’s
an optional eyepiece ranging from 5x – 30x. objective so that it
There are three types of eye lenses: can be seen by the
-Huygenian eyepiece( negative eyepiece) in human eye.
simplest form both eye and field lenses are
plano-convex, with the convex side facing the
specimen.
-Both lens and field lens of Ramsden
eyepiece (positive eyepiece) are also plano-
convex. However, its field lens is mounted
with the curved surface facing toward the
eye lens.
- Modified simple lens, also known as the
Kellner eyepiece, contains a doublet of eye-
lens elements cemented together. Unlike
Ramsden or Huygenian eyepiece, it features
a higher eyepoint and a much larger field of
view. https://www.olympus-lifescience.com/en/microscope-resource/primer/anatomy/oculars/
2 Illuminating bulb Illuminating bulbs are typically located in the The purpose of the
base of the microscope. It varies in shapes or illuminator is to
sizes. The bulbs used can be projection bulbs, provide even, high
festoon bulbs, fluorescent ring bulbs, intensity light at the
halogen, tungsten bulbs, and mercury vapor place of the field
bulbs. aperture.

https://www.microscope.com/accessories/bulbs

3 Iris diaphragm Iris Diaphragm is located below the Its main function is to
condenser and above the light source. control the amount
According to the Merriam-Webster of light that reaches
dictionary, it is an adjustable diaphragm of the specimen. It also
thin opaque plates that can be turned by a plays a major role in
ring to change the diameter of a central defining image
opening usually to regulate the aperture of a contrast and
lens. resolution.

https://www.olympus-lifescience.com/en/
microscope-resource/primer/java/dic/
opticalsectioning/

4 Condenser Condenser is a glass lens located beneath the The function of the
stage. These are lenses that are used to condenser is to
collect and focus light from the illuminator gather wavefronts
into the specimen. from the microscope
light source and
A basic condenser is fixed while an Abbe concentrate the light
condenser is movable and more precise. into the specimen.

https://zeiss-campus.magnet.fsu.edu/articles/basics/opticalsystems.html#:~:text=On%20upright
%20%20microscopes%2C%20the%20%20condenser,intensity%20%20over%20the%20entire%20viewfield.

5 Scanning objectives A scanning objective lens provides the lowest It allows the
(SO) magnification power of all objective lenses. It researchers/students
is the shortest type of objective lens, it can to observe a good
only magnify 4x. overview of the
entire slide and
The name “scanning” objective lens comes sample.
from the fact that they provide observers
with enough magnification for a wide
overview of the slide, essentially a “scan” of
 the slide.
https://sciencing.com/functions-objective-lenses-6470088.html

6 Low power The low-power objective lens has more It magnifies objects
objectives (LPO) magnification power than the scanning and provides more
objective lens; it has 10x the power of the detailed observation
eyepiece. The total magnification for this lens than SO.
is equal to 100x magnification (10x eyepiece
lens x 10x LPO = 100x magnification).

7 High power The high-powered objective lens is also It magnifies objects

objectives (HPO) known as the “high dry” lens. It magnifies 40x and allows
and its total magnification is 400 ( 10x researchers/students
eyepiece x 40x HPO = 400x magnification). to have more
detailed observations
on a slide/specimen
than LPO.

8 Oil Immersion Oil Immersion Objective lens is the longest This objective lens is
objective (OIO) objective lens. It also provides the most helpful to achieve a
powerful magnification. However, the greater clarity of
refractive index of air and your glass slide are image at high
slightly different, so a special immersion oil magnification by the
must be added to bridge the gap. Without help of oil.
immersion oil, the 100x lens will not function
correctly. The specimen appears blurry, and
you will not achieve an ideal magnification or
resolution. The total magnification of OIO is
1000x ( 10x eyepiece lens x 100x OIO =
1000x magnification).

3. Describe the principle involved in the following microscopes. Specify the uses of
each in the study of microbes.

MICROSCOPES PRINCIPLE/S INVOLVE USE/S IN

 MICROBIOLOGY

1 Brightfield The principle works by allowing a uniform It’s commonly used

microscope beam of illuminating light to pass through a to visualize, and
specimen to create an image. From the stage, identify different
microbes such as
the condenser enables the light to focus on
bacteria and
the specimen. Then with the light passing
parasites in a dark
through it, the objective lens will transmit the image against bright
contrasting image (dark image against bright background.
background) towards the eyepiece. (Mokobi,
2022) ex. To visualize and
study the
Mokobi (2022) stated that the specimen morphologies of
initially stained for contracting bacterial cell and to
characterization, it will have a refractive identify parasitic
index that will differentiate it from the protozoans such as
surrounding, presenting a combination of Paramecium.
absorption and refractive contrast. (Mokobi, 2022)

2 Darkfield Aryal (2022) describes its principle, which It is used to observe

microscope enables a specimen’s image to be viewed in a the motility of
dark field (or background) by using an flagellated bacteria
opaque disc to block off light source, causing and protozoa, and it
light to scatter as it hits the specimen. is used to exhibit very
thin bacteria (the use
On this microscope, the dispersed light or
of this microscope
zeroth order is removed so that only the
enables them to
scattered beams hit the specimen. These
appear larger). It is
light rays will hit the specimen at different
also used to observe
angles. The result is a “cone of light” where
and study marine
rays are diffracted, reflected and/or refracted
organisms such as
off the object. This will show us a contrasting
algae, plankton,
image of a bright image against dark
diatoms, insects,
background. (Aryal, 2022)
fibers, hairs, yeast
and protozoa. (Aryal,
2022)

3 Phase-contrast When light passes through cells, small phase Since this microscope
microscope shifts occur, which are invisible to the human doesn’t require the
 eye. In this microscope, these phase shifts cells to be killed, fixed
are converted into changes in amplitude, or stained, the
which can be observed as differences in specimen can be
image contrast. Also, it improves the contrast
visualized in its
of images of transparent and colorless natural state.
specimens. (Aryal, 2022) (Scientifica, n.d.)
When observing
This microscope is used ideally for thinner microbes, this can
samples (using its inverted microscope help us observe
system), and enhances the contrast of images biological processes
of transparent and colorless specimens. and specimen details
(Scientifica, n.d.) at high contrast.

4 Differential Ibidi (n.d.) describes that the principle of DIC It has the advantage
interference- microscopy has a contrast-enhancing of no phase ring in
contrast (DIC) technique which uses polarized light to the objective and a
microscope convert phase delays into intensity changes larger depth of field
(contrast). when observing a
specimen. It is used
The effect is called differential, because to detect and
contrast is created only in neighboring areas. characterize complex
On its imaging, adjacent structures with bacterial cells. It
different refractive indices are contrasted allows us to achieve a
when in close contact with each other. DIC clear visualization of
needs low birefringence of the microscopy transparent samples
chamber and the lid, making it incompatible and biological
with standard polystyrene cultureware. (ibidi, processes. Aside from
n.d.) contrast, it also
provides brilliant
pseudo-3D relief
shading images.
(Obara et. al., 2013)

5 Flourescence It uses fluorescent dyes (fluorophores or Aryal (2022) states

microscope fluorochromes) in staining the components that it offers the most
of a specimen. It uses higher light intensity to popular method for
excite fluorescence species in the sample, the study of dynamic
which then emits light of a longer behavior in live cell
wavelength. Light of the excitation imaging. It also allows
 wavelength is focused on the specimen multi-color staining
through the objective lens. The image which is helpful for
produced is based on the second light source allowing multiple
or the emission wavelength of the types of the molecule
fluorescent species. (Aryal, 2022) to be monitored at
once. With its
fluorescence, it also
assists in easily
identifying structures
of cells and the
measurement of its
physiological state.

6 Confocal Elliot (2019) states that with confocal Confocal microscopy

microscope microscopy, the illumination and detection allows 3D live
optics are focused on the same diffraction- imaging without
limited spot, which is moved over the sample physically sectioning
to build the complete image on the detector. a specimen.
Moreover, it also
Mokobi (2022) also stated that the image is
allows us to observe
formed when the microscope scans (by laser)
the changes that
the focused beam across selected area with
occur on a specimen
the control of two high-speed oscillating
such as changes in
mirrors (assisted by galvanometer motors).
microbial population
One mirror moves the beam from left to right
during a process. It
(X-axis) while the second mirror translates
also enables us to
the beam along Y-axis. After a scan on the X-
examine specimens,
axis, the beam moves quickly back to the
such as to determine
starting point to start a new scan (flyback).
its physiological
Moreover, the images are collected in a
status, without
computerized system as data which can be
extensive preparation
measured and quantified.
processes. (Takeuchi
& Frank, 2001)

7 Two-photon Ibidi (n.d.) explains that the principle is based It provides live cell 3D
microscope on fluorophore excitation (just as imaging of thick
fluorescence), which results in the emission specimens that can
of light. visualize deep
molecules, tissue
 When photons combine energy, it allows slices, organoids, etc.
low-energy infrared photons to excite Moreover, it is used
standard fluorophores. The infrared light to study host-
penetrates tissue more deeply and it’ll lead immune interactions
to excitation in the focus of the objective, in living tissues, as
because only in this area, the critical number well as to observe
of photons per time and space is reached. how pathogens such
Finally, the image is produced through laser as viruses, bacteria
scans in which it records the image intensity and parasites that
point by point (just as confocal). infect another
organism. Herz
(2012)

8 Scanning Acoustic Alter Technology (n.d.) states that this With the speed of
microscope technique is based on the reflection that sound through the
acoustic waves experience at the interface specimen correlated
between different media and density with elasticity, it
irregularities. It utilizes focalized ultrasound could conveniently
(MHz to GHz) pulses at short distances to indicate its
image material interfaces and boundaries as biochemical
well as hidden physical damages. properties, and
changes in the to
Maev (2016) added that ultrasonic image is determine their
digitally constructed from a series of A-scans functions. The
acquired during lateral scanning (mechanical observation ranges
motion of a single acoustic lens along the are shown in a
sample surface). As a result, it generates 3-D colored column.
data about the micromechanical properties Thus, precise
of the object in this particular volume. differences in
acoustic properties
are readily
distinguished by
narrowing the range.
(Miura, 2016)

9 Transmission It almost shares the similar principle to the It is widely used to

electron light microscope. However, it provides an study, visualize and
microscope image through the use of electron beams. observe cell
When the electron illuminates the specimen, structures of bacteria
 the resolution power increases hence (including its flagella
increasing the wavelength of electron and plasmids), viruses
transmission. It uses condenser lens system and fungi. Moreover,
to focus the beams on the specimen. The it is helpful in
image is displayed by objective lens which diagnostic
are further magnified by projector lenses. bacteriology such as
Moreover, it uses freeze-itching for the in determining
treatment of microbial cells. (Mokobi, 2022) bacteria and/or virus
in biopsy samples. It
is also used to locate
antigens both and
within bacterial cells.
On researches, it
proved helpful in
characterization of
motility, adhesion,
plasmid transfer and
sporulation of
bacteria. (Curry et al,
2006)

10 Scanning electron Mokobi (2022) describes that this microscope Golding et al (2016)
microscope works with applying kinetic energy to explains that SEM is
produce signals on the interaction of the used to study and
electrons. These electrons are secondary observe the filament
electrons (has a role in detecting morphology structures of different
and topography of specimen), backscattered microorganisms.
electrons (show contrast in composition of Moreover, it is also
the specimen’s elements), and diffracted used in diagnosis of
backscattered electrons which are used to infectious diseases,
view crystallized elements and photons. observation of
infectious cycles and
visualization of
microbe’s
morphological
features.

11 Scanning tunneling Quate (2009) states that the principle is It is used in analyzing
microscope based on the quantum mechanical submicron details on
 occurrence known as tunneling, in which the biological surfaces, as
wavelike properties of electrons permit them well as studying its
to “tunnel” beyond the surface of a solid into characterization
regions of space. The sharp tip of a tungsten without causing
needle is positioned a few angstroms from damage or
the sample surface. Then a small voltage is interference in the
applied between the probe tip and the sample. (Firtel &
surface, causing electrons to tunnel across Beveridge, 1995)
the gap. As the probe is scanned over the
surface, it registers variations in the
tunneling current, and this information can
be processed to provide a topographical
image of the surface.

12 Atomic force Mokobi (2022) explains that this microscope It is useful in

measures intermolecular forces and sees observing the atoms
atoms by using probed surfaces of the in a sample. It helps
specimen in nanoscale. It functions with the in its identification,
application of its 3 principles: surface sensing, evaluation of force
detection and imaging. interactions between
the atoms, and in the
The underlying principle of AFM is that this
study of its physical
nanoscale tip is attached to a small cantilever
changing properties.
which forms a spring. As the tip contacts the
(Mokobi, 2022)
surface, the cantilever bends, and the
bending is detected using a laser diode and a
split photodetector. This bending is indicative
of the tip-sample interaction force. (Mokobi,
2022)

4. Define the following properties of the microscope:

TERMINOLOGY DEFINITION

a. Magnification Magnification is the ability of a microscope to

produce an image that makes it possible to
visualize more (and closer) details of an object
too small to be seen by the naked eye.
b. Focal length The focal length of a lens system is the distance
from the lens center to a point where parallel
rays are focused on the optical axis (often
termed the principal focal point).

c. Working distance Working distance is the distance between the

front of the objective lens and the surface of the
specimen or slide coverslip at the point where
the specimen is completely in focus.

d. Revolving power The resolving power of a microscope is to

distinctly reveal adjacent structural detail. It is
this effect of diffraction that limits a
microscope's ability to resolve fine details

e. Numerical aperture Parfocal is the ability of a microscope to stay

relatively in focus as the user switches among
the objectives.

f. Parfocal It is the ability of a microscope to stay relatively

in focus as the user switches among the
objectives.

g. Depth of focus It’s the depth of specimen layer which is in sharp

focus at the same time, even if the distance
between the objective lens and the specimen
plane is changed when observing and shooting
the specimen plane.