MODULE 1: BASIC MICROSCOPY

Lesson 1 - Proper Care and Maintenance of Microscope

Proper care and maintenance of microscope is of paramount
importance and can help extend its life.
1. Handle with Care
Improper handling is a common cause of many problems that occur with
microscopes. When carrying a microscope, hold it by the base and the
metal support arm. The stage on a microscope is the flat plate where the
slides are placed for observation. Avoid picking your microscope up by the
stage or the eyepiece holder, as this can cause misalignment.
2. Look after Lenses
When using your microscope, the objective lens is lowered to adjust the
focus. However, be careful not to let the lens touch the slide you’re looking
at, as this can damage the lens. Furthermore, dirty lenses are notoriously
difficult to clean.
Avoid touching the lenses of the microscope.
3. Keep Covered
Microscopes should always be sold with dust covers. Whether transporting
or storing your instrument, make the most of the microscope bag and
remember to keep your microscope covered when not in use. The
microscope’s eye tubes also need to be kept dust free. If the eyepieces need
to be removed, cover the tubes with caps and store them with the
microscope.
4. Store Safely
Ensure you store your microscope in a clean, dry space with good
ventilation. Salt air or damp, for example, can cause damage to equipment
over time. Expensive, precision equipment should not be stored next to
solutions that may leak. Similarly, keep your microscope away from areas
with potentially corrosive chemical fumes. Such fumes can destroy lenses or
corrode metal parts.

5. Keep Clean
Oil immersion is a technique used to increase the resolving power of a
microscope. Both the objective lens and sample are immersed in a
transparent oil of high refractive index so that high magnifications can be
achieved while still maintaining good resolution. It is essential to ensure
careful cleaning takes place immediately after using immersion oil and do
not use damaging solvents.
6. Take Care of Bulbs
After using your microscope, turn off the illuminator and wait for it to
cool down before putting it away. Allowing the bulb to cool will extend its
life and avoid the unnecessary cost of expensive replacements. Similarly, if
used constantly on full power, the bulb will overheat and blow.
7. Clean Carefully
Microscope lenses are delicate. Treat them carefully to avoid any scratches.
Use an aspirator to remove dust. Moisten special lens paper with distilled
water or appropriate cleaning solution. Rubbing gently in a circular motion
will remove any sticky residue.
Rotate the nosepiece of the microscope all the way down to its
lowest level when you have finished using the microscope.
8. Refer to the User’s Manual
Your microscope should be sold with a user’s manual and specialist spanners
as required. Always refer to the manual when making any adjustments to the
microscope and use the supplied spanners.
9. Annual Maintenance Check
An annual maintenance check of microscopes is always a good idea. Moving
parts should be cleaned and lubricated. Similarly, inspect the power cords
and plugs for safety.
10. Consider a Professional Service
The company where microscope was bought can be contacted for a range of
products to help keep your laboratory microscopy equipment in tip top
condition.

Lesson 2 - Parts of the Microscope and Its Uses

Microscope "optical instrument which by means of a lens or lenses
magnifies and renders visible minute objects or details of visible bodies,"
from Modern Latin microscopium, literally "an instrument for viewing what
is small" see micro- + skopion. "means of viewing," from skopein "look at."
Microscopic "of minute size" is attested from 1760s.
The following are elemental parts of a typical microscope used in the
laboratory classes:
1. The Eyepiece Lens
Eyepiece contains the ocular lens which you will be looking
through to see the magnified specimens with magnification raging from 5X
to 30X, but 10X or 15X is the most common in use. The ocular lens provides
a re-magnified image to see when light enters through the objective lens.
2. The Eyepiece tube
It connects the eyepiece and ocular lens to the objective lenses.
3. The Microscope Arm
It connects the eyepiece tube to the base where you should hold
when carrying the microscope.
4. The Microscope Base
It provides stability and support for the microscope in its upright
position. Typically, it holds the source of light or illuminator.
5. The Microscope Illuminator
It is a light source which can come in a form of a built-in, low
voltage illuminator light, or a mirror that reflects an external light source like
sunlight.
6. The Stage and Stage Clips
It serves as the platform for slides which hold the specimen in
place through stage clip on either side.
7. The Microscope Nosepiece
It contains the objective lenses. You can rotate this part in switching
objective lenses and adjust the magnification power.
8. The Objective Lenses
Objective lenses are combining with the eyepiece lens to increase
magnification levels. Objective lenses are the lenses that protrude downward
over the specimen.
a. scanning lens – 4X
b. LPO – 10X
c. HPO – 40X
d. OIO – 100X
9. The Rack Stop
It prevents users from moving the objectives too close to the slide.
10. Control Focus Knobs
Turning the knobs, adjusts the distance between the stage and the
lens. The coarse adjustment knob is used to bring the specimen into initial
focus -- visible but not sharp. The fine adjustment knob is then turned to
bring the specimen into sharp focus.
11. The Condenser Lens and Diaphragm
These parts are located under the microscopic stage. Condenser
concentrates the light on the specimen whereas diaphragm with a small
movable lever is adjusted that regulates the entry of light.

Lesson 3 - Mounting of Specimen on Microscopic Slide

Preparation of Microscopic Slide
Mounting of specimen on slides can be done in three different ways as
discussed below in simple ways.
Dry mount slide
 1. Place sample seed of sesame on a clean glass slide at the center and
cover it.

Wet mount slide

1. Place a drop of water at the center of the slide.
2. Using tweezer, place pollen of gumamela flower or any flower with
pollen in the middle of the drop.
3. While holding the cover slip upright and by the edges, carefully place
one edge of the cover slip next to the water.
4. Set one edge against the slide and lower it until it contacts the liquid.
Slowly lower the upper edge of the cover slip onto the water.
5. Absorbent towel can be placed at the edge of the cover slip to draw
out some of the water, further flattening the wet mount slide.
Staining a Slide
Starch slurry will be mounted on a slide.
1. Place one drop of methylene blue stain or iodine solution on one edge
of the cover slip and the flat edge of a piece of paper towel on the
other edge of the coverslip. (The paper towel will draw the water out
from under the coverslip and the cohesion of water will draw the
stain under the coverslip.)
2. If the stain has covered the area containing the specimen, you are
done. (The stain does not need to be under the entire coverslip. If the
stain does not cover the area needed, get a new piece of paper towel
and add more stain until it covers.)
3. Wipe off the excess stain with a paper towel so it will not mess up
with objective lenses.
4. The slide can now be placed on the microscopic stage.

Lesson 4 - How to Focus Image Under Microscope

How to Focus Image on Your Microscope:
The following are shared steps on how to use microscope effectively while
on the actual classroom setting:
1. Place the microscope on a flat, level, firm table free from vibration
seeing to it that the arm is towards you while the stage is going away
from you. Do not place it in front of a brightly lit window.
2. Turn on the light source and adjust the optimum light setting to
ensure correct level of brightness by turning or sliding the brightness
adjustment knob at the base.
3. Turn the revolving nosepiece (turret) so that the lowest power
objective lens is clicked into position.
4. Place the slide (dry mount, wet mount, prepared mount) on
the mechanical stage secured with clips.
5. Look through the eyepiece and move the focus knob until the image
comes into focus.
6. Adjust the condenser and light intensity for the greatest amount of
light.
7. Move the microscope slide around until the sample is in the center of
the field of view.
8. Manipulate the focus knob and readjust the condenser and light
intensity to obtain the clearest image possible.
9. Once the image is sharp with LPO, move to HPO and do minor
adjustments using fine adjustment knob.
Do not let the objective lens touch the slide!
Both eyes should be open when viewing through the microscope.
10. Complete your drawings (image seen under LPO, HPO).
11. When finished, lower the stage, click the low power lens into
position and remove the slide.
12. Wash and dry the slides and keep it on your storage rack or tray.