Studypool: @laceyair

THE MICROSCOPE

WHAT IS A MICROSCOPE?
● A microscope is an indispensable
tool used in all Biological Sciences.
● This tool enables us to see plant
structures that are too small to be
seen by the unaided eye.
● Regardless of the kind of
microscope being used, three
elements are needed to form an
image: a source of illumination,
the specimen to be observed, and a
system of lenses to focus the
illumination on the sample and to
create an image.
BACKGROUND HISTORY
● Hans and Zacharias Janssen - first
compound microscopes (2 lenses),
tube with lenses at each end.
● Robert Hooke - compound
microscope improvement;
discovery of the “cell”.
● Antone van Leeuwenhoek -
created the simple microscope; is
the father of microbiology.
inclination screw, body tube, ocular
tube or draw tube, revolving
nosepiece, dust shield, adjustment
screws, coarse adjustment screw,
fine adjustment screw, stage, and
the mirror rack)
● Magnifying parts are those parts
concerned with the enlargement of
the specimen. (e.g. ocular or
eyepiece, objective: scanner, low
power objective LPO, high power
objective HPO, and the oil
immersion objective OIO)
● Illuminating parts are those parts
concerned with the light provision
and regulation of the specimen.
(e.g. mirror, diaphragm, and
condenser)
THE COMPOUND MICROSCOPE

COMPOUND LIGHT MICROSCOPE

● Is a microscope that uses natural
or artificial light and a series of
lenses: the condenser, the
objectives, the intermediate
lenses, and the ocular lens.

PARTS OF THE MICROSCOPE

● There are three basic parts of the
microscope, namely: (a)
mechanical parts; (b) magnifying
parts; and (c) illuminating parts
● Mechanical parts are those parts
concerned with the support and
adjustment of the optical parts.
(e.g. base, pillar, handle or arm,

1
Studypool: @laceyair

1. Ocular or Eyepiece magnification of

● Found on the draw tube 10-12X.
through which the operator c. High Power
peeps during actual focusing; Objective (HPO) -
usually carries a carries a
magnification of 10x. magnification of
● The POINTER is a thin 40-60X.
black line that appears to d. Oil Immersion
cut halfway across the field Objective (OIO) -
of view which the student or carries a
instructor can use to point magnification of
out regions of the 97-100X; requires
specimen under observation the use of cedar oil.

2. Ocular Tube or Draw Tube 7. Stage Clip or Specimen Holder

● The upper smaller end of the
body tube bearing the
eyepiece or ocular lens.
3. Head
● Houses the optical parts in
the upper part of the
microscope.
4. Arm or Handle
● The curved metallic part
arising from the pillar used
for holding the microscope.
● Holds the slide in place.
8. Stage
● A square or round platform
with an opening at the center
where the slide is placed.
9. Aperture
● The hole in the stage
through which the base
(transmitted) light reaches
the stage.

5. Revolving Nosepiece 10. Fine Adjustment Knob

● The circular structure where ● Is used for delicate
the objectives are focusing in connection with
attached; permits the shifting the high power and oil
of objectives. immersion objectives.

6. Objective Lenses 11. Coarse Adjustment Knob

● A tube-like structure ● Is used to adjust the low
attached to the revolving power objective in focusing.
nosepiece.
● Has 4 magnifications: 12. Light Source or Illuminator
a. Scanner - carries a ● This is the light source for a
magnification of 4-5X. microscope, typically
b. Low Power located in the base of the
Objective (LPO) - microscope.
carries a

2
Studypool: @laceyair
13. Stage Adjustment Knob
● Adjusts the position of the
mechanical stage vertically
and horizontally.
14. Condenser
● The lens found immediately
beneath the hole of the
stage.
● Is used to concentrate light
rays on the specimen.
15. Base
● Stand that supports the
microscope.
MORE PARTS OF THE MICROSCOPE
● Pillar is a short piece of metal that
attaches to one end of the base;
also supports the microscope.
● Inclination Screw is found at the
junction of the pillar and the handle
used for tilting the microscope.
● Body Tube is a cylindrical structure
vertically arising from the handle;
holds the dust shield and
nosepiece.
● Dust Shield is a circular structure
above the nosepiece used to
protect the lenses of the
objectives.
● Adjustment Screws are two pairs
of wheels attached to either side of
the body tube.
● Mirror Rack is found below the
stage and attached to the pillar;
holds the mirror in place.
● Mirror is found below the stage near
the base; is used to collect and
direct light to the specimen.
● Diaphragm is found below the
stage; used to regulate the entry of
light onto the specimen.
○ There are different types of
diaphragms: iris, plate, or
fan.
● For a demonstration of the parts and
their functions (0-7:00); for
techniques in focusing the
microscope (6:50-onwards); and
cleaning the microscope
(20:00-onwards), link to this site:
https://www.youtube.com/watch?v=o
MASHb4VyyU
HOW TO USE THE MICROSCOPE
● Focusing the Microscope
○ Care needs to be taken to
ensure that the objective lens
never strikes the stage or the
slide.
○ Focus on the low power
objective first and then move
to high power.
○ Only the fine adjustment
knob should be used when
the high power objective is
employed.
○ An oil immersion lens should
never be used without the
use of oil.
● Cleaning the Microscope
○ Lenses and eyepiece should
be cleaned with the help of
silk cloth and cleaning liquid
before use.
1. Not included in the parts of the
microscope above is the field
diaphragm. What is its function and
how is it used?
- Field Diaphragm is
responsible for controlling
the area of the circle of
light illuminating the
3
Studypool: @laceyair

specimen; hence its eradicating the possibility of

usefulness in adjusting the damaging the objective
microscope’s alignment. Due lenses.
to it being expensive, it is not
available in all microscopes; LETTER E UNDER THE MICROSCOPE
however, if present, it is
found at the base region of ● Letter E under Scanner (4X)
the microscope.

2. What is the use of the oil for the oil

immersion objective?
- Adding this oil increases the
resolving power (resolution)
- denotes the smallest detail
that a microscope can
resolve when imaging a
specimen - through the ● Letter E under Low Power
means of reducing light Objective LPO (10X)
refraction given that the
Immersion Oil has a similar
refractive index to the cover
glass.

- The refractive indices of air

and glass are different,
meaning that some of the
light bends which causes a
distortion in the image. This ● Letter E under High Power
is noticeable in objectives Objective HPO (40X)
that possess a very high
magnification power like the
Oil Immersion Objective.

3. In storing your microscope, what

should be in place? Why should this
be done?
- In storing the microscope, it
is important to always start
and end with the scanner ● The position of the letter e under a
objective in place because microscope isn’t the same when
the mechanical stage cannot viewed with the naked eye, rather it
hit the scanner objective is inverted.
even at its highest point
therefore completely

4
Studypool: @laceyair
● When the slide is moved to the left,
the direction of the letter e image
appears to move to the right.
● When the slide is moved toward
you, the direction of the letter e
image appears to move away from
you.
● When the objective is shifted to
HPO, the image becomes more
magnified and a little blurred; the
field of view also becomes
smaller.
● The microscope has convex
lenses which are responsible for
the inverted magnified images
being displayed.
● The movement of the microscope
is opposite to how it is
manipulated by the student or
instructor due to its two sets of
lenses (ocular and objective).
● The blurriness of the image as the
objective lens is switched to another
lens with a high magnification power
will depend on how it is focused
on the previous lens. If the user
was able to properly focus the
specimen on the Low-Power
Objective, for example, then the
image of the specimen won’t be so
unclear when the lens is switched to
High-Power Objective. If the user
still has a clear image of the
specimen or sample upon switching
lenses, then the user was able to
properly focus the specimen. This is
because the lenses are designed
to be Parfocal - the ability of a
microscope to stay relatively in
focus as the user switches among
objectives.
● The field of view - the size of the
area that the lens views - is smaller
because of the higher magnification
power; hence, a specific area of the
specimen or sample is being
focused on by the lens. It is
important to center the specimen
before increasing the magnification
to prevent the observer from losing
sight of the specimen or sample.
MEASURING AN OBJECT (WITH A
MICROMETER)
● An ocular micrometer serves as a
scale or ruler; it is calibrated
against a fixed and known ruler
called the stage micrometer.
● Stage micrometers come in varying
lengths, but most are 2mm long
and subdivided into 0.01mm (10
micrometers) lengths.
● Each objective will need to be
calibrated independently.
● To use, simply superimpose the
ocular micrometer onto the stage
micrometer and note the relationship
of the length of the ocular to the
stage micrometer.
● Using an ocular micrometer is the
most accurate way of measuring
the size of cells. You need to
calibrate the ocular micrometer
using a slide micrometer. Then find
the value of 1 ocular division for
each objective used, from scanner to
oil immersion objective.
5
Studypool: @laceyair

MEASURING AN OBJECT (WITHOUT A Do the same for HPO and OIO

MICROMETER) e. Measure the size of cells,
● Field of View (FOV) is the diameter which can be estimated by
of the circular image seen through the following formula:
a microscope.
Width of the cell = Field of View
Number of Cells Across
● Standard Values of Field of View:
Objective Diameter of Magnification ● Demonstration here:
FOV
https://www.youtube.com/watch?v=E_Ni
4X 4.0mm 40X yAt7pRM

10X 2.0mm 100X MEASUREMENT OF MAGNIFICATION OF

IMAGES
40X 0.4mm 400X ● Magnification of images may be defined
as the number of times an object is
100X 0.2mm 1000X enlarged by the magnifying lens or
the number of time a drawing or image
is enlarged or reduced from the original
● Finding FOV Under Scanner: size of the object.
a. Place the transparent plastic ● To determine the total magnification of
ruler on the stage so that the the specimen as seen through the
ruler’s edge is centered on microscope, multiply the
your field of view under magnification of the ocular by the
scanner. magnification of the objective.
b. Position the ruler so one of
Ocular Magnification = 10X
the millimeter markings is
just visible to the left of your Objective Objective Total
Magnification Magnification
circle in your FOV.
c. Count the number of the Scanner 4X 40X
whole millimeters that you
LPO 10X 100X
see and estimate the
fractions of millimeters. HPO 40X 400X
d. It becomes difficult to
OIO 100X 1000X
accurately estimate the
diameter of the FOV when it
switches to a higher ● To determine the magnification of
drawing:
objective. The calculation we
perform to determine the
Size of Drawing/Actual Size of Object
FOV of the higher power:

LPO (100X) = Length

Scanner (40X) x

(40)(4.55mm for example)=180 mm

180mm/100=1.8mm