Lab -1

Microscope with prepared slides

1.1 Objectives:
The objective of this lab session is to familiarize users with the operation of a microscope
by identifying its various components and understanding their functions. Users will learn how
to calculate magnification, observe changes in the field of view, and effectively locate
specimens using both low and high power objectives. Additionally, this session aims to
demonstrate proficiency in identifying slides under the microscope.

1.2 Introduction
A microscope is an instrument that is used to magnify small objects. Some microscopes can
even be used to observe an object at the cellular level, allowing scientists to see the shape of
a cell, its nucleus, mitochondria, and other organelles. While the modern microscope has many
parts, the most important pieces are its lenses. It is through the microscope’s lenses that the
image of an object can be magnified and observed in detail.
A microscope slide is a thin flat piece of glass, typically 75 by 26 mm and about 1 mm
thick, used to hold objects for examination under a microscope. Typically the object is mounted
on the slide, and then both are inserted together in the microscope for viewing.

1.3 Literature Review

Microscope are of various types such as compound microscope, light microscope and
electron microscope. A simple light microscope manipulates how light enters the eye using
a convex lens, where both sides of the lens are curved outwards. When light reflects off of an
object being viewed under the microscope and passes through the lens, it bends towards the eye.
This makes the object look bigger than it actually is.
The compound microscope, which consists of at least two lenses, was invented in 1590 by
Dutch spectacle-makers Zacharias and Hans Jansen. Some of the earliest microscopes were also
made by a Dutchman named Antoine Van Leeuwenhoek.
Leeuwenhoek’s microscopes consisted of a small glass ball set inside a metal frame. He became
known for using his microscopes to observe freshwater, single-celled microorganisms that he
called “animalcules.” Its magnification is 1000X
An electron microscope is a microscope that uses a beam of electrons as a source of
illumination. They use electron optics that are analogous to the glass lenses of an optical light
microscope to control the electron beam, for instance focusing them to produce magnified
images or electron diffraction patterns. It can magnify things up to millions of times. Its main
types are TEM and SEM.
 SEM creates an image by detecting reflected or knocked-off electrons,
 TEM uses transmitted electrons (electrons that are passing through the sample) to create
an image.
1.3.1 Parts of microscope:
Head: The head is a cylindrical metallic tube that holds the eyepiece lens at one end and
connects to the nose piece at other end. It is also called a body tube or eyepiece tube
Arm: This is the part connecting the base to the head and the eyepiece tube to the base of the
microscope.
Base: The base is the lowermost part of the microscope that supports the entire microscope
structure. It provides stability for the microscope.
Eyepiece: The eyepiece (ocular Lens) is closest to the viewer’s eye. They are located at the top
of the microscope. This part is used to look at the specimen. These lenses come in different
magnification powers from 5X to 30X, but the most common ocular lenses are of 10X or 15X
magnification. They magnify the image for the second time.
Eyepiece tube: It’s the eyepiece holder. It carries the eyepiece just above the objective lens.
Nose piece: A nose piece is a movable circular structure that houses all the objective lenses. It
is also called the revolving turret. It is connected to the body tube and lies just above the stage
Objective lenses: The objective lens is the lens that is closest to the specimen. They are fitted
on the nosepiece. A standard microscope has 3 to 4 objective lenses of different magnifying
powers: 4X, 10X, 40X, and 100X. The objective lenses first receive the light transmitted from
the specimen and magnify the image for the first time.
The Adjustment knobs: Adjustment Knobs are the control knobs used to focus the
microscope on the specimen.
 Fine Adjustment Knob: Fine Adjustment Knob is used for fine adjustment. It is a
smaller knob and is used to move the stage up or down very slowly.
 Coarse Adjustment Knob: Coarse Adjustment Knob is used for focusing the image
under low power magnification. It is a larger knob and is used to move the stage up or
down very rapidly. The stage is raised or lowered rapidly with the help of a coarse
adjustment knob.
Aperture: This is a hole in the microscope stage through which the transmitted light from the
source reaches the stage.
Condenser: These are lenses that are used to collect and focus light from the illuminator into
the specimen. They are found under the stage next to the diaphragm of the microscope. They
play a major role in ensuring clear, sharp images are produced with a high magnification of
400X and above.

1.3.2 Types of microscope slides:

There are several types of microscope slides and preparations. These include:
Dry mount: A specimen is placed on a slide with or without a coverslip over it. This method is
useful for viewing dry specimens such as hair or pollen.
Wet mount: A specimen is placed on a drop of water on the slide, with a coverslip placed over
it. This is often used for viewing cells.
Prepared slides: This type of slide has already been professionally prepared, and can last long-
term. Many types of prepared microscope slides can be purchased.
Smears: A smear is a sample that is spread across a slide and allowed to dry. A coverslip may
be used in this type of preparation. This technique is helpful for viewing blood samples.
1.4 Methodology:
Prepared slides are very easy to use since they are already professionally made. A specimen
on a prepared slide has been permanently fixed and preserved, usually through heat-fixation,
mordant application, and the use of a mounting medium. A mounting medium keeps the
specimen preserved and adhered to the slide and coverslip. It is important to carefully pick up
the slide from the sides to avoid fingerprints from getting on it. The following is method of
examining a slide on a microscope:
 Begin by ensuring that you wear a pair of gloves to prevent contaminating the samples.
Additionally, wipe down the microscope with a soft, dry cloth to remove any dust or
debris.
 Place the microscope 10cm from the end of the working table and connect the
microscope with a power connector.
 Make sure to cover your specimen that is on the glass slides with the coverslip.
 Secure your slide using the metal clips and position the specimen in the center under the
lowest objective lens.
 Use the coarse adjustment knob to bring the specimen into focus through the eyepiece,
taking care to not allow the slide to come into contact with the lens.
 Set the condenser to its highest setting to allow for maximum light transmission.
 Use the fine adjustment knob to bring the specimen into sharper focus. Be gentle when
adjusting the focus, as turning the knob too quickly or too hard can damage the
mechanism.
 Examine your specimen. Select the appropriate objective lens to view your samples.
They allow you to magnify the sample at different levels, typically 4x, 10x, 40x, and
100x. Choose the correct objective lens based on the size and nature of the sample and
the level of magnification required. Avoid touching the lenses. The lenses on a
microscope are delicate and can easily be damaged by fingerprints or other contaminants

1.4.1 Calculating total magnification power of microscope:

Magnification of microscope = Magnification of eyepiece lens × Magnification of Objective

lens. Magnification of ocular/eyepiece (10X) times magnification of objective lens (40X) =
400X

1.5 Results:
Figure 1.1: shows onion epidermis

Air space Epidermis

layer

Figure1.2: shows the onion epidermis layer on microscope

1.6 Conclusion:
Through this lab, we've learned how microscopes work and how to use them effectively.
Remember to always handle the microscope with care and practice proper slide preparation
techniques. We have learned how to calculate magnification of microscopes and analyzed
different parts of microscope.

1.7 Questions
1. Label the diagram.

Figure 1.3: shows the labelled diagram of microscope

2. What is microscope, identify its need?
A microscope is a scientific tool that magnifies tiny objects or specimens, making them
visible to the human eye. Its need arises from the necessity to explore and study the intricate
details of small organisms, cells, and structures that are otherwise invisible. It play a crucial role
in various fields such as biology, medicine, materials science, and forensics, enabling
advancements in research, diagnosis

3. Difference between light microscope, compound microscope and electron microscope?

Light microscopes use visible light to illuminate and magnify specimens, suitable for
observing living cells and tissues. Compound microscopes are a type of light microscope that
employs multiple lenses to achieve higher magnification, ideal for detailed examination of
smaller specimens. Electron microscopes, on the other hand, use beams of electrons instead of
light, offering much higher magnification and resolution, suitable for viewing ultrafine details of
specimens, but they cannot observe living specimens.

4. What is immersion oil and when it used in microscope?

Immersion oil is a type of transparent oil, typically with a high refractive index, used in
microscopy to improve the resolution and clarity of the image. It is applied between the
specimen and the objective lens. Immersion oil is primarily used with high-power objectives,
typically 40x or higher magnification, to minimize the loss of light resulting in sharper and
more detailed images

5. Why we do orientation of microscope horizontally and vertically?

Orientation of the microscope both horizontally and vertically ensures proper alignment
for accurate observation and manipulation of specimens. Horizontal adjustment allows for the
movement of the stage, enabling precise positioning of the specimen under the objective lens.
Vertical orientation ensures the correct height adjustment of the objective lens to focus on the
specimen, ensuring optimal clarity and resolution.
6. What are the effects of magnification at different(X) low power and high power?

At low power magnification, a wider field of view is observed, allowing for the
visualization of larger areas of the specimen. However, the level of detail is lower compared to
high power magnification. Conversely, at high power magnification, a narrower field of view is
seen, but finer details of the specimen become more visible due to the increased magnification.
Therefore, while low power magnification provides an overview of the specimen, high power
magnification offers enhanced resolution for detailed examination.
7. What is the effect of light intensity on sample?

The intensity of light directly impacts the visibility and clarity of the sample under the
microscope. Insufficient light may result in a dim or dark image, making it challenging to
observe fine details. Conversely, excessive light intensity can cause glare or overexposure,
obscuring the specimen's features. Therefore, adjusting the light intensity appropriately ensures
optimal contrast and clarity, facilitating accurate observation and analysis of the sample.
8. What are the safety precautions before and after working with microscope?
Before working with a microscope, it's essential to ensure the workstation is clean and
organized, free of clutter that could obstruct movement. Additionally, inspect the microscope
for any damage or defects, and ensure it’s properly plugged into a grounded outlet.
After using the microscope, turn off the light source and unplug the microscope to prevent
electrical hazards. Clean the lenses and stage with a lens paper or cloth to remove any residue
or oil. Properly store slides and specimens in designated containers to prevent contamination or
damage
9. Take two different samples for study.

Sample 1 Leaf epidermis Sample 2 Onion epidermis

Figure 1.5: onion epidermis

Figure 1.4: leaf epidermis

Leaf Epidermis:
1. Shape: Leaf epidermis cells appear irregular in shape with uneven edges.
2. Structures: Some leaf epidermis cells may have stomata (tiny pores) and guard cells that
regulate gas exchange and transpiration.
3. Patterns: Depending on the type of leaf, you may observe various patterns such as veins and
trachoma’s (hair-like structures) on the surface.
Onion Epidermis:
1. Shape: Onion epidermis cells are typically rectangular or square-shaped with clear
boundaries.
2. Layers: Onion epidermis often shows multiple layers of cells stacked on top of each other.
3. Appearance: The cells of onion epidermis are densely packed and usually lack any distinct
structures like stomata or veins

1.9 References:
 https://www.toppr.com/guides/biology/microbiology/microscope-types-uses-parts/
 https://www.microscopeworld.com/p-3658-types-of-microscopes.aspx
 https://www.microscopeworld.com/t-parts.aspx
 https://sciencing.com/magnification-microscope-5049708.html