Name: Group: Date:

Course and section:

Experiment 1: The Microscope

Objective: To know the parts and the correct technique of using the microscope.

Materials: Microscope, glass slides, prepared slide.

Observation:

The compound microscope is a light microscope that makes use of two sets of lenses
namely:
1. Primary objectives: lenses that are immediately next to the specimen being studied.
2. Secondary objectives: the oculars or eyepiece.

Observation: Draw the compound microscope and label the parts.

Questions for Research:

1. What are the names of primary objectives and give their corresponding
magnification?

 Scanning Objective Lens (4x)

The smallest magnification offered by any objective lens is that of a scanning
lens. The term “scanning” objective lens refers to the fact that it gives spectators
roughly enough magnification for a good overview of the slide, or a “scan” of the
slide. A 4x scanning objective lens produces a total magnification of 40x when
paired with a 10x eyepiece lens, which is a common magnification for scanning
objectives.
 Low Power Objective (10x)
In comparison to the scanning objective lens, the low power objective lens
has a higher magnification power. A low power objective lens and a 10x
eyepiece lens together have a total magnification of 100x, which allows
you to see the slide more closely than with a scanning objective lens while
still remaining at a safe distance for general viewing.
 High Power (40x) Objective Lens
For observing minute features within a specimen sample, a high-powered
objective lens, often known as a “high dry” lens, is perfect. You can see a
very detailed image of the specimen on your slide thanks to the 400x total
magnification that a high-power objective lens and a 10x eyepiece
provide.
  Oil-immersion objective lens (100x)
The most potent magnification is provided by the oil immersion objective
lens, which, when used with a 10x eyepiece, may achieve a staggering
total magnification of 1000x. However, because the refractive indices of air
and your glass slide are not exactly the same, a special immersion oil is
required to fill the gap. The oil immersion objective lens won’t work
properly without a drop of immersion oil added.
 Specialty Objective Lenses (2x, 50x Oil, 60x, and 100x Dry)
The 2x objective, which is frequently used in pathology, has only one-half
the magnification of a 4x scanning lens, giving the sample on the slide a
better overall view. The gold standard for seeing blood smears is the 50x
oil immersion objective, which is frequently substituted for the 40x
objective. The 60x objective, which is frequently offered in dry or oil
immersion, has 50% more magnification than a 40x lens. For larger
magnification without the use of immersion oil, the 60x dry lens is
occasionally preferred over a 100x oil immersion lens. Finally, the 100x
dry objective does not require immersion oil to produce high magnification
(still 1000x when used in conjunction with 10x eyepieces).

2. Give the functions of the following:

a. Course adjustment knob

A rapid control which allows for quick focusing by moving the objective lens or
stage up and down. It is used for initial focusing.

b. Fine adjustment knob

A slow but precise control used to fine focus the image when viewing at the higher
magnifications.

c. Stage

The flat platform that supports the slides. Stage clips hold the slides in place. If
your microscope has a mechanical stage, the slide is controlled by turning two
knobs instead of having to move it manually. One knob moves the slide left and
right, the other moves it forward and backward.

d. Condenser

Located beneath the stage and serves to gather wave fronts from the microscope
light source and concentrate them into a cone of light that illuminates the
specimen with uniform intensity over the entire view field.

e. Iris diagram
 Adjustable shutter which allows you to adjust the amount of light passing through
the condenser. The angle determines the Numerical Aperture (NA) of the
condenser. This diaphragm, generally called the aperture diaphragm, is one of the
most important controls on the microscope.

f. Illuminator

There is an illuminator built into the base of most microscopes. The purpose of the
illuminator is to provide even, high intensity light at the place of the field aperture,
so that light can travel through the condenser to the specimen.

3. What are the two types of Electron Microscope?

 Transmission electron microscope (TEM)

The transmission electron microscope is the original type of electron microscope,
which directs a high voltage electron beam towards the specimen to illuminate it and
create a magnified image of the sample. The most significant limitation of the
transmission microscope is the requirement for very thin specimen samples, usually
less than 100 nm. As a result, most biological specimens need to be chemically fixed
and dehydrated, in order to be embedded in a polymer resin so that it can be viewed
with a TEM.
 Scanning electron microscope (SEM)
The scanning electron microscope used a technique known as raster scanning to
produce magnified images of the specimen. It directs a focused electron beam across
the rectangular area of the specimen, which loses energy as it passes through. The
energy is converted into other forms of energy, such as heat, light, secondary
electrons, and backscattered electrons. This information can be translated to view the
topography and composition of the original specimen.

4. Give at least 5 ways to take good care of the microscope.

 When carrying your microscope, always use two hands

Most microscope problems occur because of improper handling so when carrying

your microscope, hold it by the bottom of the base with one hand and hold the metal
support arm with the other. Do not pick it up by the specimen stage or the eyepiece
holder, as this can cause a misalignment of lenses. When transporting it, use a
microscope bag or box and always handle it with care.

 Keep lenses clear of slides

When using your microscope and adjusting the focus you will need to lower the new
objective lens down as far as it will go. Then use the coarse and/or fine adjustment
focus to move it up until the image is focused. However, you should never allow the
lens to touch the slide you are looking at. When using the microscope, always use
the lowest power objective lens first and then increase the power as needed. Slowly
adjust the focus and keep your eyes on the distance between the tip of the objective
lens and the slides on the stage. When you using a high-magnification objective lens,
 use the fine focus instead of the coarse focus to bring your slide into focus. Dirty
lenses can be difficult to clean and the slide may leave a permanent scratch on the
surface of the lens. Once you’ve finished using the microscope, ensure to switch to
the lowest objective lens and double-check before you start a new microscope
session.

 After using immersion oil, clean the lens immediately

If using immersion oil to increase the resolving power of a microscope, always ensure
the objective lens are cleaned immediately after use. Objective, eyepieces, and
condenser may be removed for cleaning. Use only good-quality lens paper and lens
cleaning solution and do not use solvents that could damage the lens coating.
Remember to check the lens is suitable for oil immersion before using them for the
immersion oil practice.

 Always clean carefully and with the appropriate cleaning solution

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. Never use anything abrasive on microscope lenses.

 Always use the dust cover when the microscope is not in use

All microscopes are sold with dust covers so keep your microscope covered when
not in use even if the microscope is stored in a cabinet. Eye tubes also need to be
kept free of dust so do not store a microscope without its eyepieces. If the
microscope eyepieces must be removed, cover the tubes with caps or a plastic bag
with a rubber band around the eye tube.