AGR 2202 – Agricultural Microbiology

Lab # 1

Amelia Sankar – AGR1101

Title: Microscopy and parts of a compound microscope

Aim: To learn to use a microscope in the laboratory
Objectives:

• Identify and state the function of the parts of a microscope

• Properly care for a microscope
• Observe specimens using a compound microscope
Do's and Don'ts

• Always carry a microscope with both hands, one grasping the handhold in the back and one
grasping the bottom.

• Do not swing the microscope at arm’s length and do not bang it onto the bench top.

• Never place the microscope near the bench's edge and keep electric cords out of the way, towards the center of the
bench.

• All of our compound microscopes are par focal, meaning that the objects remain in focus as you change from one
objective lens to another. Examine your material first using the lower power objective (i.e. 10X); then use a
higher power objective (i.e. 20X or 40X). Because the objectives are par focal, you need to use only the fine focus
knob to fine tune your image.

• Never use the coarse adjustment to focus downward. Replace and remove a slide only after the lowest power
objective has been rotated into viewing position.

• Never attempt to repair a microscope or force an adjustment knob. You may severely damage the instrument.
Parts and Function
OCULAR LENS or EYEPIECE — on a binocular scope there are two ocular lenses, one for each eye. These lenses
magnify the image at 10X power. The power of the ocular lens multiplied by the objective lens gives the total
magnification of the microscope.
ARM — A support for the upper portion of the scope. It also serves as a convenient carrying handle.
MECHANICAL STAGE CONTROLS — A geared device to move the slide (placed in the slide clamp) precisely.
COARSE ADJUSTMENT KNOB — a rapid control which allows for quick focusing by moving the objective lens or
stage up and down. It is used for initial focusing.
FINE ADJUSTMENT KNOB — a slow but precise control used to fine focus the image when viewing at the higher
magnifications.
BASE — the part of your microscope that sits on a level, stable support.
OCULAR ADJUSTMENT — an adjustment for differences in the focusing abilities of your eyes.
DIOPTIC ADJUSTMENT — A horizontal adjustment of the oculars. Adjust for your eyes so you see only one field of
view with both eyes open.
NOSEPIECE — a circular plate with 4 objective lenses that can be rotated into position for different
magnifications.
OBJECTIVE LENS — four separate lenses that magnify the image (4X, 10X, 40X and 100X) depending on the
objective in use. The lens is positioned just above the object being viewed.
SLIDE CLAMP — clamp to hold the slide on the stage.
STAGE — platform for placement of the microscope slide.
CONDENSER — lens that concentrates or directs the light onto the slide.
IRIS DIAPHRAGM CONTROL — lever (or rotating disk) that adjusts the amount of light illuminating the slide. Use
just enough light to illuminate the object on the slide and give good contrast.
FILTER HOLDER — blue filter rests in this holder below the substage condenser.
CONDENSER HEIGHT CONTROL — A knob that controls the height of the condenser.
LAMP — the light source.
LAMP SWITCH — turns the lamp “on” and “off”
Answer the following Questions
1. What is the defining feature of the compound light microscope?
2. What is the operational principle of a microscope?
3. How is the total/effective magnification of the compound light microscope determined?
4. Briefly explain the importance of the microscope in the field of microbiology.
5. Which objective lens requires immersion oil and why?
6. Which part/(s) of the microscope should you hold when lifting or moving same?

I. A compound light microscope is a microscope with more than one lens and its own light source. This type of
microscope consists of ocular lenses located in the eyepiece and objective lenses that rotates closer to the
specimen.

II. The basic functioning principle of a microscope is that when a sample is lined into its field of focus, a resolute
and enlarged image is produced at the shortest distance from the lens that allows for distinct vision with its
magnification.

III. It is found that a compound microscope is used for its high magnification for viewing samples at high
magnification (40 - 1000x), which is achieved by the combined effect of two sets of lenses: the ocular lens (in the
eyepiece) and the objective lenses (close to the sample).

If in the case you would want to calculate the total magnification produced by the compound light microscope you
will have to:
The total magnification = multiplying the magnification of the ocular lens by the magnification of the
objective lens.

IV. There can be found many microorganisms existing around us and are known to be difficult to see or study with the
naked eye. As such the use of a microscope is intended to use because of its high magnification levels. By
identifying microorganisms, it helps broaden our understanding of life on Earth because it enables scientists to
investigate the anatomy, morphology, and behavior of microorganisms. This knowledge is essential for
understanding their biology, ecology, and interactions with other organisms in respect to microbiology.

V. The Oil immersion is a technique used typically to improve resolution of the microscope lenses and also to protect
its sensitivity when on high magnification, specifically the 100x lenses. To do this procedure the 100x lens is
placed carefully in a drop of oil that is placed on the slide while trying to ensure there is no air bubbles that would
cause loss of light “due to refraction (bending of the light) as the light passes from glass (slide) → air → glass
(objective lens).”

VI. It is advised that, when you are moving the microscope, your palm to be firmly placed at the bottom of the base,
and that your other hand to be steadily placed on the arm (which serves as the carrying handle).
Reference
• Parts of a Compound Microscope. (n.d.). MicroscopeMaster.
https://www.microscopemaster.com/compound-light-microscope.html

• Raveendran. B, (2023). Types of microscopes: definition, working principle, diagram, applications, FAQs.
BYJUS. https://byjus.com/physics/types-of-microscope/

• Biology, P. B. V. a. M. L. O. M. (n.d.). Compound and Stereo- microscopes - Microscopes 4 Schools.

https://www2.mrc-
lmb.cam.ac.uk/microscopes4schools/microscopes1.php#:~:text=Compound%20Microscopes&text=The%20total
%20magnification%20is%20calculated,allow%20this%20to%20happen%20efficiently.

• Libretexts. (2020). 17.2: Introduction. Biology LibreTexts.

https://bio.libretexts.org/Courses/College_of_the_Canyons/Bio_221Lab%3A_Introduction_to_Microbiology_(Bu
rke)/17%3A_Smear_Prep_and_Simple_Stains/17.02%3A_Introduction#:~:text=100x%20lens%20and%20Oil%2
0Immersion&text=This%20requires%20a%20special%20100x,%E2%80%8B%20glass%20(objective%20lens)