INTRODUCTION TO BRIGHTFIELD MICROSCOPE
 simplest type of compound microscope is the
Brightfield compound microscope.
 word “compound” from Brightfield Compound
Microscope means “specimen positioned properly on
the stage of a microscope and illuminated by a light
source will be magnified by two-lens system.”
 word “brightfield” means “magnified objects appear
as dark objects against a bright background.” A
sufficient contrast must exist between the magnified
object and the brightfield background for the objects to
be visible.
THE PATH OF LIGHT
 OCULAR LENS- image of the specimen is
magnified
- As “eyepiece”
 OBJECTIVE LENS- light rays
 CONDENSER-lenses that direct the light rays
 ILLUMINATOR- light source
IMPORTANCE OF USING IMMERSION OIL
 immersion oil has the same refractive index as the glass
slide
 IO is used, light rays do not refract
 resolving power of the lenses.
 When oil is not used with an oil immersion objective
lens, the image becomes fuzzy, with poor resolution.
HOW TO APPLY?
 oil used in oil immersion is “CEDAR WOOD OIL”.
 refractive index is: 1.516.
PARTS OF THE MICROSCOPE
Structural Components
1.HEAD/BODY-houses the optical parts
2.ARM- supports the microscope head
- carry microscope
3.BASE-supports the microscope and houses the illuminator
Optical Components
1. EYEPIECE/OCULAR- remagnifies the image formed by the
objective lens; standard magnifying power is 10x.
2. EYEPIECE TUBE- holds the eyepieces in place above
objective lenses.
3. OBJECTIVE LENSES- primary lenses magnifies specimen.
4. NOSEPIECE-rotating turret
5. COARSE ADJUSTMENT KNOB- bigger wheel used to adjust
the LPO in focusing; initial focusing.
6. FINE ADJUSTMENT KNOB- smaller wheel for final focusing
of the specimen using HPO and OIO; make the specimen
more vivid.
7. STAGE- slide/specimen is placed for focusing.
8. STAGE CLIPS- hold the slide in place
9. ILUMINATOR- light source
10. CONDENSER- collect and focus the light from illuminator
11. IRIS DIAPHRAGM- controls the amount of light
12. CONDENSER FOCUS KNOB- moves the condenser up or
down to control the lighting focus on the specimen.
OBJECTIVE LENSES
1. SCANNER: Magnification: 4x
2. LOW POWER OBJECTIVE (LPO):
-Magnification: 10x
-forms general outline or wider portion of the object.
3. HIGH POWER OBJECTIVE (HPO):
-Magnification: 45x
-longer than the LPO and it forms a bigger image if the
object in focus.
-enlarge specimens are small under LPO.
4. OIL IMMERSION OBJECTIVE (OIO):
-HIGHEST DEGREE OF MAGNIFICATION
-Magnification: 100x
-examine stained smear preparations of microorganisms
using immersion oil as their medium.
Calculating the Total Magnification
TOTAL MAGNIFICATION = Magnifying power of the eyepiece x
Magnifying power of the objective used
Magnifying power of the eyepiece = 10
Magnifying power of the objective used = 100 (OIO)
Correct Manipulation of Microscope
1. Place the specimen slide on the stage and secure it with the
stage clips. Arrange the portion of the slide to be examined
over the central opening stage.
2. Rotate the low power objective into place under the body
tube. You will feel a “click” when it is correctly place.
3. Raise the condenser as high as it will go.
4. Rotate the coarse adjustment knob clockwise to bring down
the LPO close but not touching the slide, until the specimen
is seen through the eyepiece.
5. Regulate the intensity of light by opening or closing the iris
diaphragm
6. Sharpen the focus by turning the fine adjustment knob.
7. Turn the LPO to HPO without elevating the body tube.
Notice that it will be almost in focus because most
microscopes are parfocal. Little adjustments with fine
adjustment knob will only be needed to clearly view the
object in focus.
NOTE: The HPO has a smaller aperture than the LPO, hence
it will be necessary to open the iris diaphragm further in
order to fill the objective aperture with light
8. To focus the oil immersion lens, swing the oil immersion
objective halfway towards the specimen in focus. Place a
drop of immersion oil on the smear. Swing the oil immersion
lens in place. Using the coarse adjustment, bring the
objective down until it touches the oil. Find the object by
gently turning the coarse adjustment knob upwards.
Sharpen the focus with the fine adjustment knob.
9. Compare the relative sizes from those seen under the LPO,
HPO, & OIO.
10. Clean the stage and the lenses after each use, before
returning the microscope to the stockroom.