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B Laboratory
C ● Pre-Focusing Knob
Lesson 1: Microscope - This knob is used to make a rough
adjustment of the focus before using the fine
Microscope adjustment knob
● Observation Tube Clamping
- ne of the most important tool utilized in clinical
O - This knob is used to tighten the eyepiece
microbiology tube in place
- To determine the cellular characteristics of ● Revolving Nosepiece
microorganisms which are helpful in their identification - A turret that holds the objective lenses. It can
➔ Compound Bright-Field Microscope be rotated to select different magnifications
- One of the most useful in clinical bacteriology ● Objective Lens
- These are the lenses that are closest to the
● Zacharias Janssen specimen
- Dutch lens maker - They provide the primary magnification of the
- Credited to have the earliest microscope microscope
● Robert Hooke ● Microscope Frame
- An english scientist who observed different - This is the main body of the microscope
microorganisms using the compound - It supports all of the other parts
microscope ● Tension Adjustment Ring
● Anton van Leeuwenhoek - This ring is used to adjust the tension of the
- 1676 eyepiece tube
- Father of microbiology - This prevents the tube from wobbling when
- Began to work on germs through his you are using the microscope
innovative single lens microscope ● Coarse Adjustment Knob
- He was able to describe tiny organisms - This knob is used to make a large adjustment
(animalcules) of the focus
➔ Animalcules - It is used to bring the specimen into general
- Derived from the Latin word focus
animalculum which means small ● Filter Holder
animal - This is a holder that can be used to insert
filters into the light path
Parts and its Function - Filters can be used to change the color of the
light or to reduce glare
● Stage Clip
- Also known as specimen holder
- This is a device that holds the specimen in
place on the stage
● Fine Adjustment Knob
- This knob is used to make a small adjustment
of the focus
- It is used to bring the specimen into sharp
focus
● Specimen Holder Y Axis
- This is the axis that allows the specimen
holder to be moved up and down
● Specimen Holder X Axis
- This is the axis that allows the specimen
● Eyepiece
holder to be moved left and right
- This is the lens that you look through to see
● Power Cord
the magnified image of the specimen
- This is the cord that is used to connect the
- It usually has a magnification of 10x or 15x
microscope to an electrical outlet
● Binocular Observation Tube
● AC Adapter
- This is a tube that holds two eyepieces so
- This is a device that converts the power from
that you can see a three-dimensional image
the electrical outlet to the voltage that the
of the specimen
microscope needs
● Diopter Adjustment Ring
● Main Switch
- This ring is used to adjust the focus of the
- This switch is used to turn the microscope on
eyepiece so that you can see a clear image
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and off
● Stage Actual Field of View
- his is the flat platform on which the
T - Size of the specimen actually observed in the
specimen is placed microscope
● Condenser
- This is the flat platform on which the 𝐹𝑖𝑒𝑙𝑑𝑁𝑢𝑚𝑏𝑒𝑟
𝐴 𝑐𝑡𝑢𝑎𝑙𝐹𝑖𝑒𝑙𝑑𝑜𝑓𝑉 𝑖𝑒𝑤 = 𝑂𝑏𝑗𝑒𝑐𝑡𝑖𝑣𝑒𝑀𝑎𝑔𝑛𝑖𝑓𝑖𝑐𝑎𝑡𝑖𝑜𝑛
specimen is placed
● Brightness Adjustment Knob
- This knob is used to adjust the brightness of WORKING
OBJECTIVE TOTAL
MAGNIFICATION ISTANCE
D
the light LENS MAGNIFICATION
(mm)
● Aperture
- This is a diaphragm that controls the amount Scanner 4x 27.8 40x
of light that passes through the condenser
● Iris Diaphragm LPO 10x 8.0 100x
- This is a diaphragm that controls the size of
HPO 40x 0.6 400x
the light beam that passes through the
aperture OIO 100x 0.13 1000x
Objective Lenses and its Magnification
Procedure
1. A lways carry the microscope in an upright position
with two hands
- One hand should support the base and the
other should be on the arm
2. Always set your microscope on a clean, flat surface
3. Place the microscope on the bench squarely and on a
stable position
4. Study the parts of the compound light microscope
5. Turn the LED Illumination on
● Scanner - Rotating the brightness adjustment knob will
- Red band increase or decrease the brightness
● Low Power Objective - The figures around the knob represent rough
- Yellow band indication of the brightness
● High Power Objective 6. Get a prepared slide and place it on the mechanical
- Blue band stage of the microscope
● Oil-Immersion Objective 7. Adjust the interpupillary distance of the microscope to
- White and black band your personal measurement
8. Raise the condenser up to the stage and adjust the iris
Total Magnification diaphragm for proper illumination
- Obtained by multiplying the eyepiece magnification 9. Adjust the diopter to compensate for the difference of
(10x) by the objective magnification eyesight between left and right eyes of the observer
10. Focus first with the scanner to view the presence of
Resolution the specimen
- Represents how finely a lens is able to distinguish the - Do not rotate the revolving nosepiece by
details of a specimen holding the objective lens
- Mainly determined by the ability of the objective and 11. Focus with the low power objective (LPO)
scarcely related to that of the eyepieces - In focusing, turn the coarse adjustment
- The function of the eyepieces is to just magnify an carefully within the working distance (WD),
image already resolved by the objective defined as the distance between the
- In case total magnifications are the same, better objective and the specimen where precise
resolution is obtained with the combinations which focus of the specimen is obtained
have higher magnification of objective 12. Turn the fine adjustment knob until you clearly see the
object in focus
Field Numbers 13. When an image has been brought into focus with the
- The diameter of the image observed through an LPO, switch to high power objective and observe how
eyepiece, indicated in millimeters the subject remains in focus
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- his is due to the parfocal feature of the
T
objectives - when a subject is in focus with
one lens, it would be in focus with all the
other lenses
14. Once in high power objective (HPO), slightly rotate the
fine adjustment knob to clearly observe the subject in
focus
- Slight adjustment is required for clear viewing
of the slides
- For high power viewing, more light is usually
needed
15. Before engaging the oil immersion objective (OIO) in
the light path, apply a drop of the immersion oil onto
the specimen at the area to be observed
16. Rotate the revolving nosepiece to engage the oil
immersion objective in the light path and bring the
specimen into focus with the fine adjustment knob
17. After use, lower the stage and rotate the revolving
nosepiece to 90° to remove the objective to which the
oil is applied
18. Thoroughly wipe off the immersion oil from the front
lenses of the objective and the condenser with lens
paper slightly moistened with absolute alcohol
- Using paper towel will scratch the lenses.
Use a soft cloth to clean other parts of the
microscope
- Wipe off the immersion oil from the specimen
top using clean gauze
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