Microscope with Prepared Slides
1.1 Objective
This lab has the following objectives:
• To become familiar with the use of microscope, its parts, and its function
• To calculate magnification, describe changes in field of view and view specimens at
different powers.
• To observe various specimens on prepared slides under microscope
1.2 Introduction
1.2.1 History
The microscope's history traces back to the late 16th century when Dutch spectacle makers crafted
rudimentary lenses, leading to the invention of the simple microscope. In the 17th century, Antony
van Leeuwenhoek's refined lenses enabled groundbreaking observations of microorganisms. Over
centuries, advancements in optics and technology birthed compound microscopes, electron
microscopes, and other specialized variants, revolutionizing scientific exploration of the
microscopic world.
1.2.2 Overview
Microscope is a tool that produces enlarged images of small objects, allowing the observer to have
an exceedingly close view of minute structures in a slide. It is primarily used for examination and
analysis. Its key parts include the eyepiece, objective lens, stage, and illuminator, working together
to produce enlarged images of tiny specimens.
Microscopic organisms and samples require the use of a microscope to magnify an image large
enough to view. The compound light microscope is commonly used to view transparent objects
that can fit on a microscope slide. A microscope slide is a thin, rectangular glass plate that can
hold a variety of microscopic specimens. There are different ways to prepare a microscope slide
depending on the type of specimen you are viewing. The main purpose of microscope slides is to
facilitate microscopic examination. Prepared slides are placed under a microscope to study tissue
or cell samples and their structures at high magnification to determine whether they are normal or
abnormal. In this lab different specimens on prepared slides are viewed under the microscope.
1.3 Literature Review
The literature concerning this lab consists of the introduction of microscope and its parts followed
by calculation of magnification and the procedure to view a prepared slide under a microscope.
Prepared slides provide a convenient way to observe specimens.
1.3.1 Types
Microscopes come in various types, each designed for specific purposes:
1. Optical Microscopes: These use visible light and lenses to magnify samples, including
compound microscopes for viewing thin slices and stereo microscopes for three-
dimensional observation.
2. Electron Microscopes: Utilizing beams of electrons instead of light, electron microscopes
offer incredibly high resolution, including transmission electron microscopes (TEM) for
internal structures and scanning electron microscopes (SEM) for surface imaging.
3. Confocal Microscopes: These utilize laser beams to scan samples layer by layer, enabling
three-dimensional imaging with high resolution and contrast.
4. Scanning Probe Microscopes: Operating at the atomic level, these microscopes include
atomic force microscopes (AFM) and scanning tunneling microscopes (STM), allowing
for precise imaging and manipulation of surfaces at nanoscale resolutions.
5. Digital Microscopes: These modern microscopes incorporate digital cameras and
computer interfaces for capturing, analyzing, and storing images digitally, offering
convenience and versatility in research and education.
1.3.2 Microscope and its Parts
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. It is composed of:
i. Lenses
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.
ii. Eyepiece: The eyepiece, also known as the ocular lens, is where the viewer peers through
to observe the magnified image of the specimen.
iii. Objective lenses: These lenses are positioned above the specimen and provide different
levels of magnification, ranging from low to high power, allowing for detailed examination
of the specimen.
iv. Condenser
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.
v. Base
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.
vi. Stage
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.
vii. Adjustment Knobs
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.
viii. Light Illuminator
It has a light illuminator or a mirror found at the base or on the microbes of the nosepiece.
ix. Nose Piece
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.
x. Diaphragm
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.
Figure 1.1 Parts of a light microscope
1.3.2 Magnification
The compound microscope uses two lenses to magnify the specimen: the eyepiece and an objective
[Link] most microscopes, there is a choice of objectives to use. Magnification can therefore be
varied, according to the size of the specimen to be viewed and the level of detail required.
The total magnification of a microscope is:
Magnification of the microscope = magnification of eyepiece × magnification of objective
Example:
So, if the magnification of an eyepiece is ×10 and the objective is ×4, the magnification of the
microscope is:
Magnification of eyepiece × magnification of objective = 10 × 4 = 40
1.3.3 Prepared Slides
Prepared microscope slides are ready-made slides that have a specimen preserved and stained.
Specimen preservation is accomplished by applying chemicals to help affix it to the slide and
coverslip. The slides that are viewed in this lab experiment include the cycloid and cucurbit slides.
i. Cycloid
Cycloid scales are round and flexible structures which are present on the surface of bony
fishes. Cycloid scales have an outer layer of calcium and an inner layer made of connective
tissues. Cycloid scales are the inner layer of ganoid scales. Example: Trout, Herring, Carp,
etc. It exhibits the following structures under a microscope.
Figure 2.1 Structure of a cycloid
ii. Cucurbit
Cucurbita (Latin for 'gourd') is a genus of herbaceous fruits in the gourd family. The stem
of Cucurbita is having a wavy margin, there are distinct ridges and furrows found here. The
vascular bundles are found to be arranged in two rings, one ring of smaller bundles against
the ridges and one ring of larger bundles are present against the furrows.
Figure 1.3 Structure of a cycloid
1.4 Methodology
The methodology is given as follows:
1.4.1 Components
• Light microscope
• Prepared glass slides
1.4.2 Procedure
The procedure is given below.
• First of all, make sure the microscope is placed on a levelled surface.
• To achieve the best possible lighting, turn on the microscope and adjust the light
intensity.
• After the slide has been prepared, place it on the microscope stage and fasten it with
the stage clamps.
• Using the coarse adjustment knob, lower the stage until the objective lens is nearly
in contact with the slide, starting with the lowest magnification objective lens (often
4x).
• Using the coarse adjustment knob, focus the specimen roughly through the
eyepiece.
For a sharp image, use the fine adjustment knob to fine-tune the focus.
• If required, repeat the focusing procedure using higher magnification objective
lenses (10x, 40x, or 100x).
• When you change the magnification, make sure the specimen stays in focus and in
the middle.
• Make any required modifications for a given after taking note of any structures or
features that you see.
• When you're done, take the slide off the stage, adjust the objective lens to its lowest
magnification, and switch off the microscope.
1.5 Results
The results showed:
1. For Cycloid: The result typically demonstrates a curve traced by a point on the
circumference of a circle rolling along a straight line. It showcases the characteristic shape
of the cycloid, often resembling a series of arcs.
2. For Cucurbit: The outcome typically reveals the anatomical structure of cucurbit plants,
including features such as leaves, tendrils, flowers, and fruits. It may highlight details such
as cellular organization, vascular systems, and reproductive structures.
Figure 1.4 The microscopic view of cycloid
Figure 1.5 The microscopic view of cucurbit
1.6 Conclusion
In this lab the basics of a microscope are studied, its usage, its parts and the function and role of
each of its parts. The calculations for magnification are looked into and different specimens are
viewed under the microscope. The prepared slides are carefully set and observed under the
microscope. The cellular structure of cycloid scale and cucurbit stem are carefully observed and
analyzed
1.7 Questions
1. Label the diagram.
Ocular Lens
Diopter Adjustment
Head
Nose Piece
Arm
Objective Lens
Stage Clip
Mechanical Stage
Aperture
Coarse Adjustment
Diaphragm
Fine Adjustment
Condenser
Stage Controls
Illuminator
Base
Brightness
Adjustment
Light Switch
2. What is microscope, identify its need.
A microscope is an optical instrument used to magnify small objects for detailed observation. Its
need lies in its ability to reveal microscopic structures and organisms, which are otherwise invisible
to the naked eye, enabling scientific research and medical diagnosis.
3. Difference between light microscope, compound microscope and electron microscope.
Light microscopes use visible light to illuminate and magnify specimens, compound microscopes
have multiple lenses for increased magnification, and electron microscopes use electron beams for
even higher magnification and resolution.
4. What is immersion oil and when it used in microscope.
Immersion oil is a type of oil with a similar refractive index to glass. It is used in microscopy to
reduce light refraction and improve resolution, particularly when using high magnification
objectives.
5. Why we do orientation of microscope horizontally and vertically?
We orient the microscope horizontally to ensure the specimen is centered on the stage and parallel
to the objective lenses, facilitating accurate focusing and observation. Vertically, we adjust the
microscope to align the lenses with the light source and eyepiece, optimizing the optical path for
clear imaging and minimizing aberrations.
6. What are the effects of magnification at different(X) low power and high power?
At low magnification, a wider field of view allows for the observation of larger areas but with less
detail, while high magnification provides greater detail but with a narrower field of view, requiring
more precise positioning and focus adjustments.
7. What is the effect of light intensity on sample?
Light intensity affects the brightness and clarity of the sample under the microscope. Proper
illumination is crucial for optimal visualization of the specimen, as too much or too little light can
result in poor image quality.
8. Take two different samples for study.
Sample 1 : Cycloid Sample 2 : Cucurbit
Write the observation of above both samples.
The cycloid specimen when viewed under the microscope shows an anterior firld and a lateral field
in the form of circular ring like structures. Some star like structures are also observed that are the
plasma vesicles. The cucurbit on the other hand shows different structures of a plant stem like the
phloem vessels and the layers of epidermis etc.
9. What are the safety precautions before and after working with microscope?
Before working with a microscope, ensure the workspace is clean and free from clutter. After use,
turn off the microscope, clean lenses with lens paper, and return all components to their proper
places to prevent damage or contamination. Additionally, wash hands thoroughly to avoid any
potential exposure to harmful substances.
1.7 References
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