USE AND CARE OF THE MICROSCOPE

The unaided eye has the ability to distinguish between two items if they are at least 0.1mm
apart (resolving power) (Vovopich and Moore, 1999). By using a light microscope we are able to
distinguish between two items that are at least 0.1μm apart (Vovopich and Moore, 1999)
During your course of study you will need to view biological structures and organisms too small
to be seen by the unaided eye. Two types of microscopes will be needed to assist you in viewing
these specimens. They are the compound binocular microscope and the dissecting microscope.
Parts of a Microscope

1. Eyepiece/Ocular Lens – The lens into which the user looks to see the specimen.
2. Diopter Adjustment – Used to alter focus between eyepieces to
3. Arm – A supporting piece of the optical microscope mounted upon the base.
4. Nose Piece – A rotating turret for switching between objective lenses.
5. Objective Lenses – Lenses with various magnification strengths.
6. Slide Holder – Clips to keep the slide in place.
7. Stage – The platform holding the specimen slide.
8. Coarse Focus – Brings specimen into general focus.
9. Fine Focus – Fine tunes the focus of the specimen.
10. Condenser – Focuses light from the light source onto the specimen.
11. Iris Diaphragm – An opaque iris composed of blades made to pass light through an aperture.
12. Base – The supporting block of the light microscope.
13. Light Source – A light or a daylight directed via a mirror.
14. On/Off Switch – You probably understand what this does :)

STERIO MICROSCOPE/ DISSECTING MICROSCOPE

There are two basic types of microscopes: compound and stereo. Stereo microscopes use lower
power than compound microscopes do and have typical magnification levels ranging from about
10 to 40x. These microscopes are very versatile and are designed for viewing whole objects such
as rocks, insects and flowers, but can also be used for viewing prepared slides. They are a helpful
tool for nature studies.
 Steps in Using a Stereo Microscope
 Set your microscope on a tabletop or other flat sturdy surface where you will have plenty of
room to work. Plug the microscope's power cord into an outlet, making sure that the excess cord
is out of the way so no one can trip over /it or pull it off of the table.
 Switch on the light source(s). If you will be looking at a microscope slide or other transparent
object, bottom lighting will work best. If the specimen you are viewing is opaque or solid (light
cannot pass through it from below), use top lighting so that the light can reflect off the
specimen's surface.
 Center your specimen on the stage plate. If your specimen is thin and flat, or if its edges curl up
easily, try using the stage clips to hold it in place. To do this, pull up the pointed end of one stage
clip and slide it over one end of the specimen, then do the same with the stage clip on the other
side. If your specimen is larger than the stage plate, you might need to turn the stage clips out so
that they are hanging off the stage giving you more room to work.
 Adjust the eyepiece(s) so that you can look through the microscope comfortably without
straining your eyes. (See below for instructions on adjusting a stereo head). For light-colored
translucent specimens such as salt crystals, use either the black side of the stage plate (if it is
reversible) or a piece of dark-colored construction paper to provide contrast.
 If your stereo microscope has a rotating objective turret, turn it so that the marking on the
objective you want to use is facing the front of the microscope. To determine the magnification
 of your microscope, multiply the magnification level of the eyepiece lens by that of the objective
lens. For example, on the microscope in the diagram above, the total magnification at the 2x
objective is 20x (2 x 10 = 20).
 While looking through the eyepiece(s), slowly turn the focus knob until the specimen comes into
view. Once you can see the outline of the specimen, turn even more slowly to focus as sharply as
possible. If you aren't able to see anything, try moving the specimen around slightly on the stage
plate to make sure it is directly below the objective lens and then try focusing again. Once you
have focused on the specimen, you can move it around to see its other parts. You may have to
refocus slightly on each new area. Note: with a stereo microscope you will often be viewing
three-dimensional specimens that have many different levels. You will not be able to focus every
feature clearly at the same time.
 When you are finished using your microscope, turn off the switch, remove the specimen, unplug
the power cord, and cover the microscope with its dust cover. Store your microscope in a place
where it will not be damaged from extreme hot or cold temperatures, and out of the reach of
small children.
Ideas for Using a Stereo Microscope
Clear plastic or glass petri dishes are great for viewing live or messy objects with a stereo
microscope because they fit well on the stage and keep everything adequately contained. The
suggestions below are just a few things you can use petri dishes to view with your stereo
microscope. Place the item or items to be viewed in the bottom of a petri dish and position it on
the stage plate of your microscope. Use top or bottom lighting.

Cleaning and Maintenance

To clean the exterior side of lenses, use a non-solvent cleaning solution designed for cleaning
optics or eyeglasses. First remove dust with a soft brush or can of compressed air. Then moisten
a piece of dry lint-free tissue or lens paper and clean the lens surfaces with a circular motion.
Repeat with a second piece of paper moistened with lens solution if necessary. Repeat once
again with dry lens paper until the lens is clean and dry. Use this same procedure for the
eyepiece and objective lenses.
To clean the interior side of lenses, use a bulb-type duster or a compressed gas canister designed
for cameras and other optical equipment. Do not use the above procedure.

To clean the body of the microscope, use a soft dry or damp cloth. Using a dust cover will help
keep your microscope investment clean and dust-free.

A low power or stereo microscope typically employs objective lenses of 50x or less. It is used to
view specimens that are visible to the naked eye such as insects, crystals, circuit boards and
coins.
Handling the Microscope

• Always use two hands to move the microscope. Place one hand around the arm, lift the
scope, and then put your other hand under the base of the scope for support.
 • Be gentle. Setting the microscope down on the table roughly could jar lenses and other
parts loose. The microscope seems like a simple instrument, but each lens (eyepiece and
objective) is actually made up of a number of other lenses; banging your microscope
around, shakes about 15 to 20 lenses

• Always have clean hands when handling your microscope.

Storing the Microscope

– Return the lowest power objective in place
– Wrap the cord around the base
– Dust is an enemy to microscope lenses; always keep the microscope covered
when not in use.
Cleaning the Microscope
– Don’t let the microscope get too dirty – always use the dust cover when not in
use.
– To clean the eyepiece – use a high quality lens paper. First brush any visible dust
from the lens, and then wipe the lens. Do not use facial tissues, they are made
from ground up wood fibers and could damage the lenses.
– To clean the objective lenses – use a fresh piece of the lens paper each time so
that you don't transfer dust from one lens to another.
– Use lens paper on all glass parts of the microscope.
– Clean oil immersion lens with chemicals provided by your instructor

MICROMETRY/MEASURING OBJECTS UNDER THE MICROSCOPE

– Meaning of Micrometry:
– Micrometry is the science in which we have some measurement of the dimensions of
an object being observed under the microscope. The method employs some special
types of measuring devices which are so oriented that these can well be attached to
or put into the microscope and observed. The object, to be measured, is calibrated
against these scales.
– Once we are observing an object under a microscope by the 5X objective and the 10X
eyepiece we say that the image that we are able to perceive is 5 × 10 = 50 times of the
object.
– We get the magnified view no doubt and also that it is the perfect coordination of the
dimensions, but to find out the exact size of the object will need precision and that is
achieved through the application of some small scales called micrometers.
– Types of Micrometry:
– There are usually two types of micrometers, i.e. stage micrometer and ocular meter
or ocular micrometer (Figs. 104, 105).

–
–
– i. Stage Micrometer:
– As is clear from its name it is for the measurement on the stage of the microscope
where an object is to be kept. This micrometer is of a slide’s shape and size and has a
mount of very finely graduated scale. The scale measures only 1 mm and has a least
count of 0.01 mm, i.e. 1 mm region is divided into 100 divisions. As 1 mm has 1000µ,
one division of stage micrometer is equivalent to 10µ.
–

– ii. Ocular Meter:

– This micrometer is used inside the eyepiece. The upper eye lens is unscrewed and the
ocular meter is put into the tube of eyepiece, and the eye lens is again replaced in its
original position. There are usually 50 or 100 divisions in the ocular meter which are
engraved on the glass.
Measurement of the Microorganisms:
When the microscope is caliberated, then the object or organism to be measured is kept
on the sage of the microscope and is observed through the eyepiece with ocular. The
object is measured in the particular magnification by ocular divisions and then is
changed into microns by multiplying ocular divisions with calibrated value of one ocular
division in that particular magnification.