Andrea Jean Miranda BSED SCIENCE-2B

Anatomy and Physiology September 8, 2022

THE COMPOUND MICROSCOPE

Generally, cells of plants and animals are too small to be seen with the naked eye, thus,
necessitating the use of the special instrument. Such instrument is a microscope, which may be of two
general types: a) the light microscope which makes use of visible light; b) the electron microscope, which
makes use of a beam of electrons rather than visible light.
The compound microscope is an example of a light microscope that makes use of two sets of
lenses:
a) primary magnifiers, the objectives, the lenses of which are immediately next to the specimen being
studied; and b) the secondary magnifiers, the ocular or eyepiece, the lenses of which are immediately
next to the eye.
Important terms in microscopy:
1. Virtual image or floating image – image seen in the microscope field.
2. Microscope field – circular area seen through the ocular where the image of the specimen is
observed.
3. Magnification – the ratio of the apparent size of the object, as seen through the microscope, and the
actual size of the object.
a. total magnification or total power the – product of the magnifying capacity of the eyepiece and
the objective being used.
Ex. high power objective 40x
Ocular 10x
Total power 40 x 10 = 400
This means that the image is 400 times as big as the object under study.
4. Units of measurement
a. micrometer (m) – unit of measurement of a light microscope which is equivalent to 10 -6.
b. nanometer (m) – unit of measure of an electron microscope that is equivalent to 10 -9.
5. Focusing – the adjustment of the spatial relationship between the objective lens and the specimen.
When a clear image of the specimen is seen through the ocular lens, the specimen is said to be “in
focus”.
6. Focal length of focus – the distance from the optical center of the lens to the focal point.
7. Working distance – the distance between the front (bottom lens) of the objective lens system and
the top of the cover glass.
Note: Lenses of high magnification and short focal lengths have a short working distance.
8. Angular aperture – the angle or cone of light rays, capable of entering the front lens of the objectives
from the point in the object.
Note: More light rays from the specimen can be taken in by the lens if the angular aperture of the
the objective is increased, thus increasing also the resolving power.
9. Numerical Aperture (NA) of an objective – the measurement of the angle of the maximum cone of
light that may enter the lens, or simply a measurement of the ability of the condenser and objective
lens to gather light.
Objective Numerical Aperture
4x 0.10
10x 0.25
20x 0.40
40x 0.65
Note: The higher magnification of the objective, the larger the numerical aperture (NA).
10. Resolving power (RA) – the ability of the lens to clearly separate or distinguish two points or two
lines individually in the image.
11. Depth of focus – represents the thickness of the specimen that may be seen in focus at one time.
- decreases with an increase in the numerical aperture and magnification.
12. Parfocal – term applied to the objective lens when practically no change in focus has to when one
power is substituted for another.
Note: Only a slight turn of the fine adjustment is needed when a change is made from low power
to a high
power objective.
13. Refraction – bending of light away from the objective lens when light passes from the glass of the
microscope slide to the air.
Note: Refraction is lessened by using oil (oil immersion objective).
14. Cover glass – thin glass (0.17 to 0.18 mm thick) cut in circles, rectangles, or squares for covering the
specimen.

PARTS OF A COMPOUND MICROSCOPE

I. MECHANICAL PARTS:
1. Base – horseshoe or V-shaped structure that supports the entire microscope.
2. Pillar – vertical post extending from the base to the arm.
3. Arm – curved basic frame of the microscope to which the base, body, and stage are attached.
- serve as the handle for carrying the microscope as well as providing support for the body tube.
4. Inclination joint – a movable hinge that facilitates tilting of the microscope by holding the arm with
your right hand and the bases with your left hand.
5. Body tube – a hollow cylinder in front of the upper part of the arm and the serves as the optical
housing for the lenses.
6. Draw tube – the smaller cylinder attached to the upper part of the body tube and serves to hold the
ocular eyepiece.
7. Revolving nosepiece – a rotating head attached to the bases of the body tube.
8. Dust shield – a fixed head attached to the base of the body tube.
9. Stage – a platform, which may be circular or square, where the slide with the specimen is placed.
- usually equipped with a pair of clips that hold the slide in place.
- Some microscopes are provided with mechanical stage adjustment knobs for accurate
movement of the slide.
- Provided with a central opening that allows light to pass from the mirror to the object being
studied.
10. Substage condenser with iris diaphragm – a structure found below the stage holding the abbe
condenser above and the iris diaphragm below. The iris diaphragm is provided with a projecting
handle or lever which when manipulated can regulate the amount of light passing through the lenses
into the observer’s eye.
11. Adjustment knobs – consist of two pairs of knobs:

a. an upper pair, the coarse adjustment knob or screw which is used for initial focusing by
raising or lowering the body tube with the scanner or the low power objective in position.
b. A lower small pair, the fine adjustment knob or screw which is used for final viewing and
adjustment at different depths, with the high power objective in position.
Note: Never use the coarse adjustment screw when focusing on the high-power objective or any
high-powered objective. Use only the fine adjustment screw. The coarse adjustment screw can
be used when the low power or scanner objective is in place.

II. ILLUMINATING PARTS:

1. Mirror – usually two-faced, of which one surface is plane or flat while the other side is concave
(parabolic)
- held by a double-hinged mirror rack or fork which facilitates adjustments at any
desired angle since the mirror functions to reflect the light into the specimen.
- The concave mirror is used with low-intensity light which may be either natural or
artificial, while when the source of light is very bright, the plane mirror is best
used.
2. Condenser (abbe condenser) – the substage optical element which may be fixed or provided with
adjustment for lowering or raising.
- functions to condense and concentrate reflected light from the mirror through the
specimen at a precise angle to illuminate the microscopic field.

III. MAGNIFYING PARTS:

1. Eyepiece or ocular – the detachable cylinder equipped with lenses and located above the draw tube.
- usually provided with a dark pointer used to point to particular parts of the
specimen to be identified.
2. Objective – generally with two or three objectives.
a. Low power objective – the shortest cylinder or tube with a large lens opening, large
lenses, and lower magnification.
b. High power objective – the longer cylinder or tube with smaller lens openings, smaller
lenses, and higher magnification.
c. Oil immersion objective – the longest cylinder or tube with very small lenses and very
high magnification.
d. Scanner – if there are four objectives present, then this would be the shortest cylinder
with the biggest lenses and lowest magnification.

Care of the Microscope

Microscopes are expensive tools, hence, must be handled very carefully at all times. The
following are some of the precautions that you must be familiar with when using them.

a. Examine your microscope before using it for any defects, like misplaced or missing parts. If
there is
anything wrong with it, call the attention of the instructor, otherwise, you will be held
responsible
for any defects.
b. Keep it clean before and after using with a piece of soft cloth (not your handkerchief).
c. Do not separate loose parts, like the ocular or objectives, as they may collect dust.
d. Do not touch the lenses with your fingers. Use lens paper only to clean them.
e. Always check the parts thoroughly after receiving it from the counter and before returning.
f. If anything is spilled on the microscope, clean it off immediately.
g. Always carry the microscope carefully with one hand under the base and the other hand
around the
arm. Never carry the microscope hanging like a basket with only one hand.
Name/s: Andrea Jean Miranda Date Performed: __________________
Group No. _____________________________ Date Submitted:
___________________

HOW TO USE THE MICROSCOPE

Activity No. 3

This exercise affords the opportunity to become familiar with the essential principles of
microscope use. Through practice, your technique with the microscope improves.

Materials: Microscope cut out letters from the newsprint

Glass slide Small piece of coarse woven cloth
Coverslip Small beaker with water
Dropper Forceps

I. Position:
a. Place the microscope on the table with the arms toward you and the base at a safe distance
from the edge of the table so that you do not cramp your body when you look through the
eyepiece.
b. Swing the low power objective in the position directly above the stage aperture.
II. Lighting:
a. Open the diaphragm wide. Make sure that the top lens of the condenser is at the same level
as the stage.
b. Look through the ocular and adjust the mirror until you get the greatest possible amount of
light reflected in the objective. If the field of the microscope is properly lighted, you should
see a circular area of light that is uniformly bright.
III. Locating the object under the lower objective:
a. Cut out a letter “e” from the classified ad of a newspaper. Place this letter in an upright
position on a clean slide with a drop of water and cover with a cover slip.
b. Look at the objective from the side and turn the coarse adjustment knob until the tip of the
objective is about 2 mm from the cover glass. Remember not to let the objective touch the
coverslip for it may damage or break the lens of the objective or the cover slip.
c. With both eyes open, look through with one eye and slowly turn the coarse adjustment knob
until the image of the object comes into clear focus.
d. Center the object by moving the slide in the right direction and clip the other end of the slide
in place. Change the opening of the diaphragm until the field is just bright enough for you to
see the details of the object clearly. Too strong a light is hard on your eyes.
e. Look at the upright letter through the microscope under low power.
f. Bring the image to a sharper focus by turning either direction the fine adjustment knob.
g. Look at the upright letter “e” through the microscope.
Answer the following questions:
1. Is the letter right side up or upside down? Explain.
- If we put the letter "e" under the microscope, we would see an upside-down
letter "e" and backward. It is completely flipped around. It is because of the two
lenses on the microscope. Due to the biconvex lens or simply means a lens that
is curved on both sides which is the reason to refract light in such a way it will flip
the image over and turn it upside down.
magnification, the letter e appears inverted under a microscope when compared
to the letter e seen normally. When the slide moves to the right, it moves toward
the left. Thus, it clearly shows that when we move it, it also moves to the
opposite side.

2. Reverse the position of the slide. Is the letter right-side-up now?

- Yes, when we reverse the position of the slide, it gives us the letter "e" that we
see normally with our naked eye. It's because when we move the slide to the
right, it will also move toward the left. It always moves on the opposite side.

3. Now move the letter from the right to the left on the stage. Which way does the letter
seem to move as you see it in the ocular? Why is it so?
- If we move the specimen to the left, it will appear to move to the right through
the eyepiece. It is because of the curvature of the first lens, which causes it to
invert the image.

h. Once you have learned how to use the low power objective, you should have no difficulty in
using the microscope with the high-power objective. The microscopes assigned to you have
objectives adjusted by the maker so that they are parfocal. This means that when an object
can be clearly seen under low power, the objective can be shifted to the next higher
magnification with little or no change in focus.

IV. Using the high power objective:

 a. Rotate the nosepiece to bring the high power into position. If the object appears slightly
blurred, turn the fine adjustment knob very slowly until the image is in sharp focus again.
Caution: Never use the Coarse Adjustment with the High Power. Do not let the
objective lens touch
the cover glass of the slide.

Answer the following;

1. What is the difference (from that seen in the low power objective) in
a. the size of the field?
- When we use the high-power objective lens of the microscope it allows us to
see the specimen closer which shows more detailed image. Unlike in the low
power objective that we can see the whole specimen just like the letter “e”,
but if we change it to the higher objective, we will only see the specific part of
the specimen, but in more detailed.

b. The amount of light?

- In high-power objective we see less light, which the image appears dimmer.

c. The position of the letter seen?

- If we change the lenses into high power objective, it does not change the
position of the letter not until we move the specimen.

Note: 1. When starting with a microscope study, always examine the specimen under
the LPO
first before shifting to HPO.

V. Magnifying Power
The compound light microscope is used to examine details of the cellular and tissue structure of
animals. The usefulness of a light microscope is limited not by its ability to magnify objects, but by its
ability to discern small objects that are very close together. This is called RESOLUTION and is related to
the wavelength of visible light used to illuminate an object.
The microscope allows biologists to extend the vision to a resolution of fractions of a micrometer.
This degree of resolution requires sophisticated electron microscopes, not generally used by
undergraduate students. Lower magnification, however, is just as useful for most aspects of animal
biology. For undergraduate laboratory work, microscopes of magnifications ranging from 5x to 1,000x are
commonly used.
a. To find the total magnifying power of the microscope, multiply the magnification of the
objective by
that of the ocular. The magnifying powers are marked on both parts. Compute the
magnifying power
of the microscope assigned to you, using both the low power and the high power objective.
b. Magnifying power is in terms of diameters, not in areas. A 10-X lens makes a line appear to
be ten
times longer than without magnification.
c. In microscopic work, a common unit of measurement is the micron (µ). A micron is 1/1000 of
a
millimeter (1 millimeter = 1,000 µ).

Links that will help you on the microscope activities, since you cannot perform actual manipulation of
microscopes.

https://www.youtube.com/watch?v=0at-m9bXArk How to perform the “letter e”

https://www.youtube.com/watch?v=3ZFW_nbV7RM Virtual Microscope Tutorial
Name/ s : Andrea Jean Miranda Date Performed:
Group No. ______________________________ Date Submitted:

THE COMPOUND MICROSCOPE

Activity No. 2

A microscope is an essential tool in the study of the fine structure of animals. It enables you to
see the animal structures too small to be seen with the naked eye. The microscope is made up of a
system of lenses that can invert images. It is important to learn the correct use of this tool from the start
so that you realize its full potential. The accuracy and value of many of your observations in the laboratory
will depend on your skill in the use of this instrument.

I. Objectives: At the end of the period, students will be able to:

1.
2.
3.
II. Materials:
Compound microscope
Pencil
Eraser
III. Drawing: Draw the compound microscope and label the parts.
IV. Questions:
1. What are the three major parts of the microscope?
- The three major parts of the microscope are the mechanical parts, illuminating parts, and
magnifying parts.

2. What are the illuminating parts? Describe and discuss them in your own words.
- The Illuminating part of the microscope is the mirror and condenser. Mirror is located beneath
the stage and has a concave and plane surface to gather and direct light to illuminate the subject.
While the condenser is a set of lenses between the mirror and the stage that concentrates light
rays to the specimen. Thus, this part of the microscope provides low and high intensity of light
which enables us to see the details of the subject that is put under the microscope.

3. What are the magnifying parts? Describe and discuss them in your own words.
- The magnifying part of the microscope is the eyepiece and the objective. Eyepiece is another
set of lenses that is found on top of the body tube which function to further magnify the image
produced by the objective lenses. On the other hand, objective is a metal cylinder attached below
the nosepiece and contains specially ground and polished lenses. This part allows us to view,
magnify, and produce image of the specimen that is placed on the slide.

4. What are the mechanical parts? Describe and discuss them in your own words.
- The mechanical parts of the microscope are the base, pillar, arm, inclination joint, body tube,
draw tube, revolving piece, dust shield, stage, substage condenser, and adjustment knobs. This
part holds the structure together and control their use. This part is formation of the microscope
that makes us called the microscope, microscope.

5. How important is the use of the microscope and the technique’s application in your subject in
anatomy and physiology as well as a BSED Science student?
- Microscope enables us to see that things that is no visible in the naked eye. It helps us
determine what is the smallest thing in our body. Also, it displays the microscopic structure of
organs and tissues, elucidates the relationship between structure and function, provides insight
into the underlying chemistry by selective staining, and allows for the comparison of normal and
abnormal tissues. As a BSED Science student microscope plays an important field because we
are studying different things that is macroscopic and microscopic. With the help of microscopes, it
enables us to discover and learn what makes us an organism we will discover new things that
could blown our mind and amaze our eyes. Furthermore, microscopes will be the tool for us to
gain new knowledge; identify different kinds of tissue, what cells look like and other more.