ELECTRICAL ENGINEERING DEPARTMENT

BIOMEDICAL INSTRUMENT
DEU50053

PRACTICAL WORK 6
PROGRAM: DEU

STUDENT NAME STUDENT NUMBER

Nurul Lidiya Bt Mohd Sulaiman 08DEU21F1077
Nur Anisah Bt Mohd Razali 08DEU21F1108
Nur Elyana Bt Zuraiwan 08DEU21F1097

LECTURE NAME: DR. BAHARUDDIN MUSTAPHA

 PRACTICAL WORK ASSESSMENT

DEU 50053 BIOMEDICAL INSTRUMENTATION

Session : 1 2023/2024

Programme : DEU 5B Practical Work Number :PW 6

Date : Lecturer’s Name :DR BAHARUDDIN MUSTAPHA

PRACTICAL WORK ASSESSMENT GENERIC

TOTAL SKILL
A. PRACTICAL ASSESSMENT
B. LAB
NO. REG. NO. NAME GROUP MEMBERS SKILL
REPORT GSA
ASSESSMENT
ASSESSMENT
(A+B=100%)
(30%) (100%)
(70%)

08DEU21F1077 NURUL LIDIYA BT

1
MOHD SULAIMAN
08DEU21F1097 NUR ELYANA BT
2
ZURAIWAN
08DEU21F1108 NUR ANISAH BT MOHD
3
RAZALI

PRACTICAL WORK ASSESSMENT

NO A. Practical Skill Assessment Attainment B. Lab Report Attainment
( CLO3, PLO5) (1–5) Assessment (1–5)
Student able to identify, Report format
1 choose and use equipment and organization
correctly
Student able to set and Result
2
calibrate equipment correctly
Student able to construct Analysis circuit
3
circuit correctly
Student able to take Discussion
4
measurement correctly
Student able to follow Conclusion
5 instruction and procedure
correctly
Student able to complete task Reference
6
given within time frame
Student have high self- ability
7
to complete the task
TOTAL ( /35 ) X 2 TOTAL /30

= / 70
PRE-LAB QUESTION:
Learn about various types of microscope and how it does.
Stereo Microscope

What is a Stereo Microscope?

A stereo microscope is an optical microscope that provides a three-dimensional view of a specimen. It is

also known by other names such as dissecting microscope and stereo zoom microscope.

The Characteristics of a Stereo Microscope

 Two separate objectives

 Two separate optical paths
 Uses the light reflected from the object
 Typical magnification range between 10x and 50x
 Three-dimensional images

How Do Stereo Microscopes Work?

A stereo or a dissecting microscope uses reflected light from the object. It magnifies at a low power hence
ideal for amplifying opaque objects. Since it uses light that naturally reflects from the specimen, it is helpful
to examine solid or thick samples. The magnification of a stereo microscope ranges between 10x and 50x.

Additional supplementary/auxiliary lenses can be attached to increase or decrease magnification and

adjust working distance based on the user's needs. For instance, if a longer working distance is required
than a lens of less than 1x is required: 0.3x, 0.5x, or 0.7x. An ocular lens or eyepiece can allow you to
increase the total magnification of your microscope to 300x or higher.

Opaque objects like coins, fossils, mineral specimens, insects, flowers, etc. are visible under a dissecting
microscope magnification. More advanced stereo microscopes can allow you to view electrical components
and circuit boards.
Parts of a Stereo Microscope

Parts Function

Stereo Head the moveable top portion of the microscope and the stereo
head holds the two adjustable eyepieces.
Ocular Lens These are the eyepieces through which the viewer looks
at the specimen. The eyepieces are typically set at 10x
magnification. It is also possible to upgrade to a higher
magnification level.
Dioptre Setting It compensates for the focusing differences between the
left and right eye. Setting it correctly prevents eye strain.
Objective Lens Stereo microscopes have two separate objectives, each
one connecting to one of the eyepieces. The eyepiece and
the objective lenses collectively determine the
magnification of the microscope. They can have a fixed
single objective, a rotating multiple lens turret or a zoom.
It allows you to change the magnification levels depending
on the applications.
Focus Knob Stereo microscopes usually have one focus knob. Helps
move the head of the microscope up and down to bring a
sharp image of the object. Most dissecting microscopes
have standard "rack and pinion" focusing. Rack is the
track with teeth and pinion is the gear that rides on the
teeth. The knob helps the pinion move along the rack.
Stage Clips help to hold the slides or other thin objects in place on the
stage.
Stage Plate It is located directly under the objective lens. It is where
the specimen is placed for viewing. Some stereo
microscopes have reversible black and white stage plates
to provide appropriate contrast with the object being
viewed.
Illumination Many microscopes have both top and bottom lighting.
Some microscopes only have one source of illumination,
top or bottom. The top lighting shines down on the
specimen and reflects light off them. The bottom lighting
transmits light up through the stage to show translucent
specimens.
 Compound Microscope

What is a compound Microscope?

A compound microscope is a type of microscope with more than one lens. It consists of
two optics components known as an objective lens and an eyepiece or ocular lens. It is also
known as a biological microscope since it is used for laboratory purposes.

Working Principle of Compound Microscope

o The specimen or item to be studied is often mounted on a clear glass slide and placed on
the stage between the condenser and objective lenses.
o A condenser lens directs visible light from the base to the specimen.
o The objective lens collects the light emitted by the specimen and magnifies it to generate
the main image within the body tube. The ocular lens magnifies this image once more.
o When a higher magnification is necessary, the nose piece is turned after low power
focusing to align the objective of a higher power (often 45X) with the lit portion of the
slide.
o Very high magnification is occasionally necessary. Thus, an oil immersion objective lens
(often 100X) is used.
o The image can be seen through the eyepiece.
Parts of a compound Microscope

Parts Function

Base The base, often known as the foot, is either U-shaped or

horseshoe-shaped. It is a metallic framework that holds the
entire microscope together.

Pillar The pillar acts as a link between the base and the arm.

Arm The arm, also known as the limb, is a metallic handle that
connects the arm to the inclined joint. The arm holds up the
stage and the body tube.

Stage It is a metallic platform with a hole in the middle that is

attached to the bottom half of the arm. Side clips or
mechanical stage clips are used to secure the microscopic
slides to the stage for observation.

Body Tube The body tube’s primary function is to retain the objective and
ocular lenses at the two ends. The end with the ocular lens is
known as the head, while the end with the objective lens is
known as the nose piece. There is a channel for light rays to
move through the body tube under ray optics.

Drawing Tube a tiny fixed tube at the top end of the body tube. The draw
tube’s primary function is to hold the ocular lens.

Pinion and Rack The rack and pinion is either linked to the body tube or the
stage to bring the item into focus.

Screws for adjusting
These are two pairs of adjusting screws that are used for
either coarse or fine adjustment. Fine adjustments move the
body tube or stage exceedingly short distances, and coarse
adjustments move the body tube and stage higher. A crisp
image may be obtained by careful tweaking.

Manual Stopper A tiny screw on the rack and pinion is used to prevent the
body tube from slipping downhill. This protects the objective
lens from damage.
Diaphragm The diaphragm controls the quantity of light that falls on the
item. It may be found beneath the stage. The two types of
diaphragms are the disc and the iris.

Condenser It is found under the diaphragm. Light may be focused by

changing the condenser, which can be moved up or down.

Reflector a mirror mounted above the base. The mirror features a plain
mirror on one side and a concave mirror on the other. The
plane mirror side is utilized when the light is strong, while the
concave mirror side is used when the light is faint. The light
on the object is directed via the diaphragm and condenser
with the aid of the reflector.

Objective lenses These lenses are located above the nose piece. The objective
lens is a compound lens that creates a true inverted picture of
the image within the body tube.

Ocular Lens often referred to as the eyepiece. Through these lenses, the
image of minute things may be seen.
 Fluorescence Microscopy

What is a Fluorescence Microscopy

an optical microscope that uses fluorescence and phosphorescence instead of, or in addition to,
reflection and absorption to study the properties of organic or inorganic substances.

Principle of Fluorescence Microscopy

Most cellular components are colorless and cannot be clearly distinguished under a microscope.
The basic premise of fluorescence microscopy is to stain the components with dyes.
Fluorescent dyes, also known as fluorophores or fluorochromes, are molecules that absorb
excitation light at a given wavelength (generally UV), and after a short delay emit light at a longer
wavelength. The delay between absorption and emission is negligible, generally on the order of
nanoseconds.
The emission light can then be filtered from the excitation light to reveal the location of the
fluorophores.

Typical components of a fluorescence microscope are:

Fluorescent dyes (Fluorophore)
 A fluorophore is a fluorescent chemical compound that can re-emit light upon light
excitation.
 Fluorophores typically contain several combined aromatic groups, or plane or cyclic
molecules with several π bonds.
 Many fluorescent stains have been designed for a range of biological molecules.
 Some of these are small molecules that are intrinsically fluorescent and bind a biological
molecule of interest. Major examples of these are nucleic acid stains like DAPI and Hoechst,
phalloidin which is used to stain actin fibers in mammalian cells.

A light source

 Four main types of light sources are used, including xenon arc lamps or mercury-vapor
lamps with an excitation filter, lasers, and high- power LEDs.
 Lasers are mostly used for complex fluorescence microscopy techniques, while xenon lamps,
and mercury lamps, and LEDs with a dichroic excitation filter are commonly used for wide-
field epifluorescence microscopes.

The excitation filter

 The exciter is typically a bandpass filter that passes only the wavelengths absorbed by the
fluorophore, thus minimizing the excitation of other sources of fluorescence and blocking
excitation light in the fluorescence emission band.

The dichroic mirror

 A dichroic filter or thin-film filter is a very accurate color filter used to selectively pass light of
a small range of colors while reflecting other colors.

The emission filter

 The emitter is typically a bandpass filter that passes only the wavelengths emitted by the
fluorophore and blocks all undesired light outside this band – especially the excitation light.
 By blocking unwanted excitation energy (including UV and IR) or sample and system
autofluorescence, optical filters ensure the darkest background.

Application of Fluorescence Microscopy

 To identify structures in fixed and live biological samples.

 Fluorescence microscopy is a common tool for today’s life science research because it allows
the use of multicolor staining, labeling of structures within cells, and the measurement of the
physiological state of a cell.
 Light Microscope

What is light Microscope

 A light microscope is a biology laboratory instrument or tool, that uses visible light to detect
and magnify very small objects and enlarge them.
 They use lenses to focus light on the specimen, magnifying it thus producing an image. The
specimen is normally placed close to the microscopic lens.
 Microscopic magnification varies greatly depending on the types and number of lenses that
make up the microscope. Depending on the number of lenses, there are two types of
microscopes i. e Simple light microscope (it has low magnification because it uses a single
lens) and the Compound light microscope (it has a higher magnification compared to the
simple microscope because it uses at least two sets of lenses, an objective lens, and an
eyepiece). The lenses are aligned in that, they can be able to bend light for efficient
magnification of the image.
 The functioning of the light microscope is based on its ability to focus a beam of light
through a specimen, which is very small and transparent, to produce an image. The image is
then passed through one or two lenses for magnification for viewing. The transparency of
the specimen allows easy and quick penetration of light. Specimens can vary from bacterial
to cells and other microbial particles.
Principle of a Light Microscope
As mentioned earlier, light microscopes visualize an image by using a glass lens, and
magnification is determined by, the lens’s ability to bend light and focus it on the specimen,
which forms an image. When a ray of light passes through one medium into another, the ray
bends at the interface causing refraction. The bending of light is determined by the refractive
index, which is a measure of how great a substance slows the speed of light. The direction and
magnitude of the bending of the light are determined by the refractive indexes of the two
mediums that form the interface.

A medium with a lower refractive index such as glass to air normally speeds up the light
penetration and makes light bend away from the normal and when light is passed through a
medium with a greater refractive index such as air to glass, it normally slows down and bends
towards the normal, perpendicularly to the surface.
If an object is put between these two mediums i.e between water and air, in this case, a prism, the
prism will bend the light at an angle. This is how the microscopic lenses work, they bend the light
at an angle. The lens (convex) on receiving the light rays, focuses the rays at a specific point
known as the focal point (F-point). The measure of distance from the center of the lens and the
focal point is known as the focal length.
A microscope uses lenses whose strength is predetermined, in that, the strength of a lens is
directly related to the focal length i.e short focal length magnifies objects more than lenses with a
long focal length.
Microscopy works strictly with a factor of resolution whereby resolution is the ability of a lens to
be able to differentiate small objects that are closely packed together. The resolution of a light
microscope is determined by a numerical aperture of its lens system and by the wavelength of
the light it employs; a numerical aperture is a definition of the light wavelengths produced when
the specimen is illuminated.
A minimum distance (d) between two objects that distinguishes them to be two separate entities,
determined by the wavelengths of the light can be calculated by an Abbe equation using the
wavelength of the light that illuminated the specimen (Lambda, λ) and the numerical aperture
(NA, n sin Ɵ) i.e. d=0.5 λ/n sin Ɵ
Parts of a bright-field microscope (Compound light microscope)

It is composed of:
 Two lenses which include the objective lens and the eyepiece or ocular lens.
 Objective lens is made up of six or more glasses, which make the image clear from the
object
 The condenser is mounted below the stage which focuses a beam of light onto the
specimen. It can be fixed or movable, to adjust the quality of light, but this entirely depends
on the microscope.
 They are held together by a sturdy metallic curved back used as an arm and a stand at the
bottom, known as the base, of the microscope. The arm and the base hold all the parts of
the microscope.
 The stage where the specimen is placed, allowing movement of the specimen around for
better viewing with the flexible knobs and it is where the light is focused on.
 Two focusing knobs i.e the fine adjustment knob and the coarse adjustment knob, found on
the microscopes’ arm, which can move the stage or the nosepiece to focus on the
image. the sharpen the image clarity.
 It has a light illuminator or a mirror found at the base or on the microbes of the nosepiece.
 The nosepiece has about three to five objective lenses with different magnifying power. It
can move round to any position depending on the objective lens to focus on the image.
 An aperture diaphragm also is known as the contrast, which controls the diameter of the
beam of light that passes through the condenser, in that, when the condenser is almost
closed, the light comes through to the center of the condenser creating high contrast. But
when the condenser is widely open, the image is very bright with very low contrast.
DISCUSSION :
Let's study the fundamental procedures for utilizing a compound light microscope as we
delve into the interesting field of microscopy today. Gaining precise and in-depth
observations requires knowing the right ways to operate a microscope, whether you're a
biology lab student or just a keen enthusiast.

 First things first, make sure the microscope is on a firm surface and turn it on.
Then, use the various settings to adjust the light's intensity. A good source of
illumination is essential to producing high-quality observations.
 The objective lenses of the microscope are its heart. Start by choosing the lens
with the lowest magnification, which is typically marked 4x. You can rotate the
nosepiece to switch to higher magnifications as you advance.
 Let's discuss getting your specimen ready next. If needed, fasten it in place after
placing it on a spotless microscope slide. You can add a cover slip for extra
security.
 The X and Y controls on a microscope with a mechanical stage are used here to
move about and examine various areas of your material. Throughout your
observations, the mechanical stage improves control and precision.
 A critical ability in microscopy is focusing. To achieve a rough focus on your
specimen, start by turning the coarse focus knob. Next, use the fine focus slider
to sharpen the image for an incredibly detailed perspective
 For a more comfortable viewing experience, think about modifying the diopter
settings and the interpupillary distance, if applicable. These modifications
guarantee that using the microscope is comfortable for the eyes in addition to
being productive.
 Remember to swap out your objective lenses for different magnifications as
you proceed with your observation. To preserve clarity, always turn on the fine
focus knob after switching targets.
 Record your observations in a journal or on a digital device that you bring with
you. Make a note of the magnification, any noteworthy features, and any other
pertinent information. This exercise helps you gain a thorough understanding of
your specimen.

To wrap up our discussion on using a microscope, remember to turn off and unplug the
microscope after use. Cleaning the lenses and other components ensures that your microscope
remains in top-notch condition for future observations.
STEP TO CLEANING A MICROSCOPE :

Cleaning a microscope is crucial for maintaining its performance and ensuring accurate
observations. Here are step-by-step instructions for cleaning a microscope:
Materials Needed:
o Lens cleaning solution: Use a commercial lens cleaning solution or a mixture
of distilled water and isopropyl alcohol (in equal parts).
o Lens paper or lens cleaning tissue: Ensure it is clean and free of lint.
o Cotton swabs or lens cleaning pens: Use these for hard-to-reach areas.
o Compressed air: To blow away dust and debris.
o Mild detergent solution: For cleaning external parts.
o Soft brush or camel hair brush: To remove loose particles.
Cleaning Steps:
1. Turn Off and Unplug:
 Ensure the microscope is turned off and unplugged to avoid electrical hazards.
2. Remove Accessories:
 Take off any accessories, such as eyepieces and objective lenses, carefully.
3. Blow Away Loose Particles:
 Use compressed air to blow away any loose dust or particles from the lenses
and other surfaces.
4. Clean External Parts:
 Wipe down the exterior surfaces with a soft cloth or cotton swab moistened
with a mild detergent solution. Be gentle to avoid damaging labels or
markings.
5. Clean Eyepieces:
 Moisten lens paper with lens cleaning solution.
 Gently wipe the eyepieces in a circular motion, starting from the center and
moving outward.
 Use a dry portion of the lens paper to remove any remaining solution.
6. Clean Objective Lenses:
 Apply a small amount of lens cleaning solution to lens paper.
  Wipe the objective lenses in the same circular motion, taking care to avoid
touching the glass with your fingers.
 Use a dry portion of the lens paper to remove any remaining solution.
7. Clean Condenser and Diaphragm:
 If your microscope has a condenser and diaphragm, clean them using the same
method as for the lenses.
8. Clean the Stage:
 Wipe the stage with a clean, damp cloth to remove any debris.
 If necessary, use a soft brush or cotton swab to reach corners and edges.
9. Check Mechanical Parts:
 Inspect mechanical parts like the focusing knobs and stage controls. If needed,
use a cotton swab or lens cleaning pen for cleaning.
10. Inspect and Reassemble:
 Check for any spots or streaks on the lenses. If necessary, repeat the cleaning
process.
 Reassemble the microscope, ensuring all components are properly seated.
11. Store Properly:
 If the microscope won't be used for an extended period, cover it or store it in a
protective case to prevent dust accumulation.
TROUBLESHOOTING
1. Check Power Supply:
 Ensure that the microscope is properly plugged in and receiving power.
 Check the power outlet and try a different one to rule out electrical issues.
2. Illumination Issues:
 If the microscope has a light source, make sure it is functioning properly.
 Check the bulb and replace it if necessary.
 Verify that the condenser and diaphragm are properly adjusted for optimal
illumination.
3. Focusing Problems:
 If the microscope is not focusing correctly, check the objective lenses for any
debris or damage.
 Make sure the specimen slide is properly placed on the stage and secured.
 Check if the fine and coarse focus adjustments are working smoothly.
4. Lens and Eyepiece Issues:
 Inspect the eyepiece and objective lenses for cleanliness. Clean them using
lens paper and appropriate cleaning solutions if needed.
 Ensure that the lenses are properly aligned and centered.
 Check for any scratches or damage on the lenses.
5. Condenser Adjustment:
 Adjust the condenser height and aperture to optimize the illumination and
contrast.
 Make sure the condenser is centered and properly aligned.
6. Stage Movement:
 Check if the stage moves smoothly along the X and Y axes.
 Lubricate the stage if it appears to be sticking or moving unevenly.
7. Mechanical Issues:
 Inspect the microscope for any loose or damaged parts.
 Check the mechanical stage, focusing knobs, and other controls for proper
functioning.
8. Alignment Issues:
 If the images appear distorted or misaligned, check the alignment of the optics.
 Consult the microscope manual for instructions on adjusting the alignment.
9. Magnification Problems:
 Confirm that the correct objective lens is in place and functioning.
 Verify that the eyepiece is properly seated and not damaged.
10. Microscope Software (if applicable):
 If your microscope is connected to a computer, check the software for any
error messages or updates.
 Ensure that drivers are up-to-date.
11. Consult the Manual:
 Refer to the microscope's user manual for specific troubleshooting steps and
maintenance guidelines.
CONCLUSION
In conclusion, the process of becoming acquainted with biomedical equipment,
particularly the microscope, is a crucial aspect of professional development in the field.
Familiarization with this specialized equipment not only enhances one's technical skills but
also facilitates a deeper understanding of its applications in biomedical research and
diagnostics. For both inexperienced and experienced users, microscopes should always be
handled with care. Proper microscope use will help prevent damage to the equipment and
prevent laboratory accidents such as breaking slides. Optical microscopes use light
illumination, lenses, and often a digital camera to magnify images, with a typical resolution
limit of about 0.2 µm and magnification in the 1500× range for visible light. Types include
stereomicroscopes, compound microscopes, and laser-based Raman systems. As practitioners
immerse themselves in the intricacies of microscope operation and functionality, they lay the
foundation for more effective and precise scientific exploration. Ultimately, this familiarity
contributes to the advancement of biomedical sciences by empowering professionals to harness
the full potential of microscopy in their work.
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