DEPARTMENT OF ARTS AND SCIENCES

LABORATORY ACTIVITY 1
MICROSCOPY
Second Semester
AY 2022-23

Name: Program: BS Psychology

Section: BSP 1-1 Date: March 6, 2023

ACTIVITY – MICROSCOPY

Introduction to the Microscope

Overview
Follow along in this online exercise, with guided instruction covering the basics of light microscopy,
comparable to what you would learn in a seated lab. You will identify components of the microscope,
understand the functions of those components, learn how to focus in on a specimen, and review proper
care and maintenance.

Learning Objectives
 Identify the parts of the light microscope and describe the function of each.
 List the steps in focusing a light microscope.
 Describe how to properly handle the light microscope, focus slides, and clean the microscope when
finished.

Background
 When we walk out the door each day we see many examples of life on earth - trees, birds, other humans.
Unless we look closer, we will miss the huge diversity of life that is too small to be seen with our eyes
alone. The majority of life on earth is microscopic and, until tools were available to visualize these
organisms, they were completely overlooked. One of the first tools that opened our eyes to microbial life
was the microscope. The light microscope uses lenses along with light to magnify items up to 1000x.
This allows us to see organisms like bacteria, archaea, yeasts, protozo-ans, and algae that we would
never notice because they are so small. In addition, we can also look closer to see cells that are the
building blocks of the macroscopic organisms (like ourselves, the trees, and birds we noticed earlier.)
In addition to magnifying organisms, most cells are colorless so many times preparing slides also
involves adding stains (dyes) to color the cells, so they are more easily viewed. The slides you will
practice with today are either naturally pigmented or have been stained so we can find them more easily.
Look for these colors as you are focusing the virtual microscope.

PROCEDURE 1: Introduction to the Microscope and its Parts

1) Go to http://www.ncbionetwork.org/iet/microscope/.
2) Click on the Guide link (bottom of the home page).
3) Click through the six parts of the Guide, starting with the Introduction. You can use the arrows at the
bottom of the Guide box to guide you through the chapters.
4) When you have completed all six sections, click Close.
5) Next click on the Learn link (bottom of the page), which will take you to an image of a microscope
with question marks.
6) Starting at the top of the microscope, click on the question marks identifying the parts of the
microscope.
7) Read the description of the part of the microscope and take notes as needed.
8) Continue clicking on question marks until all turn to green check marks.
a. Do not forget to click on the question marks for items associated with the microscope.
b. You may click on any green check mark to review any part of the microscope.
c. Use the Microscope Parts checklist on the next page to ensure all parts have been identified.
9) Click on the Next button (bottom right).
10) Start on the left and click on the question mark. When the lens enlarges, click on each question mark
until each turns into a green check mark. Read the descriptions and take notes as needed.
11) Click on the Next button (bottom right).
12) Click on the Dry Slide and Oiled Slide buttons to see the difference in why immersion oil is used for
the 100X objective lens.
13) Click on the Next button (bottom right).
14) Click on the Eyepiece Options and Lens Options to learn about calculating total magnification. Try
all combinations and see how the Letter E slide image changes.
15) Click on the Next button (bottom right) to return to the home page.
16) Answer the provided questions on the next page.

Microscope Parts
 Identify all parts of the microscope and associated items.

K – On/Off Switch C – Eyepiece

J – Arm F – Nosepiece

G – Objective Lenses D – Stage

H – Diaphragm E – Stage Adjusment Knob

M – Base L – Coarse Adjusment Knob

A – Lens Paper N – Fine Adjusment Knob

B – Immersion Oil P – Slide Box

O – Kimwipes

Questions

1) What is the proper way to carry microscope?

To ensure the safe movement and setup of a microscope, it is crucial to handle the device with care.
 Make sure the microscope is turned off and unplugged from the power source if it has one.
 Use both hands to pick up the microscope. Place one hand under the base and the other hand
around the arm of the microscope.
 Hold the microscope firmly and lift it gently and smoothly. Avoid jerky or sudden
movements that could damage the internal components of the microscope.
 Keep the microscope level and close to your body as you move it. Do not swing it around or
carry it at an angle as this could cause the objectives to collide with other objects.
 If you need to transport the microscope over a longer distance, use a carrying case or box
designed for microscopes to provide extra protection.
 It's important not to remove the dust cover until the microscope is positioned on the table
where it will be used.

2) What is the typical magnification of an ocular lens? What other magnifications are possible?
The typical magnification of an ocular lens, also known as an eyepiece, can vary depending on
the specific instrument it is being used with. However, in most cases, the magnification range of
ocular lenses is between 5x and 30x. For example, in a typical compound microscope, the eyepiece
 magnification is usually 10x. In telescopes, the eyepiece magnification can range from 5x to 30x,
depending on the focal length of the telescope and the specific eyepiece being used.
The other magnifications are:
 Low-power objective: 2x to 10x magnification
 High-power objective: 40x to 100x magnification
 Oil immersion objective: 100x to 1000x magnification
 Eyepiece lens: 5x to 20x magnification

3) What are the magnification abilities of each of the objective lenses? What is the total magnification
with each objective?
Formula for the total magnification:
Ocular lens x Objective lens = Total magnification
a. Scanning (small lens), red ring = allowing the user to see the image through the eyepiece, this
small lens also called as "scanning lens" is useful for quickly finding an area of interest on a
specimen. Above it is its color band which is the red ring, this color band provides a visual
preference for the resolution of the lens when you can't see the magnification specifications.
Since the magnification of the lens is 4x, the given color will be Red. For the total
magnification of this lens, 4x/0.10 is the given total.
Scanning Lens (4x)
5x x 4x = 20x
10x x 4x = 40x
15x x 4x = 60x
20x x 4x = 80x

b. Low-power (medium lens), yellow ring = The medium lens also known as Low - Power lens
focuses and useful for identifying large details on a specimen and also setting up the view for
higher powered objectives. At the top of lens which is its color band or its yellow ring
provides of course the visual preference for the resolution for 10x lenses. The color yellow is
known and used for lenses with 10x magnification. The total magnification for this medium
lens or the Low - Power lens is 10x/0.25.
Low - Power (10x)
5x x 10x = 50x
10x x 10x = 100x
15x x 10x = 150x
20x x 10x = 200x

c. High-power (large lens), blue ring = ideal for observing fine details within a specimen
sample, such as nerve cells in the retina or the striations in skeletal muscle. The color blue is
used for this lens as it stands for 40x magnification.
High-power (40x):
5x x 40x = 200x
10x x 40x = 400x
15x x 40x = 600x
20x x 40x = 800x

d. Oil immersion (largest lens), white ring = is used for examining the details of individual
cells. This lens requires a special immersion oil to bridge the gap between the edge of the
objective and the cover slip. Color white is used as it denotes its resolution of 100x.
Oil Immersion (100x):
5x x 100x = 200x
10x x 100x = 1000x
 15x x 100x = 1500x
20x x 100x = 2000x

4) Why do you use immersion oil with 100X objective lens?

The immersion oil helps to refocus the light as it passes from the slide to the lens.This provides
an image with better resolution and contrast.

5) What is the total magnification of a sample with an ocular lens power of 15X and using a 40X
objective lens?
Total magnification = Objective lens
Magnification x Ocular lens magnification
Total magnification = 40X x 15X
Total magnification = 600X

6) What is a diaphragm? What does it do?

In a microscope, the diaphragm refers to a small circular or square-shaped disc with several
different-sized holes or an adjustable opening. It is located below the microscope stage and above the
light source.
The primary function of the diaphragm is to control the amount of light that passes through the
specimen on the slide. By adjusting the diaphragm, the user can regulate the amount and intensity of
light that illuminates the specimen, providing better contrast, clarity, and resolution of the image.
In addition to controlling the light, the diaphragm also helps in adjusting the depth of field. A
smaller aperture opening will increase the depth of field, while a larger aperture opening will
decrease it, enabling the user to focus on different parts of the specimen.
Overall, the diaphragm is an essential component of the microscope that allows the user to
control the light and optimize the image quality while observing the specimen.

Reference

© 2018 NCBioNetwork.org. Retrieved from: http://www.ncbionetwork.org/iet/microscope/