602 Biology

Experiment Number: 1
TITLE: CALIBRATION OF A MICROSCOPE: How to Calibrate a light microscope
without a Stage and Ocular Micrometer

Aim:
1. To calibrate x4, x10 and the x40 objective lens of a light microscope using a
graph paper.
2. To determine the width of a hair follicle using the calibrated light microscope.

Introduction:
Microscopic measurements are always of great importance to biologists as you made need to find out for
example the diameter of for e.g. a white/ red blood cell. The smallest unit of macroscopic metric
measurement is the millimeter (mm). However this is an extremely large unit for microscopic
measurement. For measuring microscopic measurement the biologist uses a unit known as micron (μ)
which is 0.001mm. Research microscopes are often equipped with an optical micrometer, a glass disk
with a micron scale which is placed in the ocular, as shown. By moving the slide, an object may be placed
along the scale and measured accurately. Without an ocular micrometer, however, you can determine the
approximate size of many subjects by comparing them with the known diameter of the low power and
high power fields.

Ocular Micrometer

Materials and Apparatus: Microscope, graph paper, slide, cover slip, strand of hair.

Procedure
Activity 1

1. To measure the diameter of the low paper field lay the graph paper across the stage opening.
Find the graph paper in the low power field.
2. Move the graph paper so that one of the vertical lines dividing the graph paper into
millimeters is just visible at the left edge of the field. The distance between the darker blue lines is
 1 cm therefore the distance between the smaller lines is 1mm. Measure the distance across
(diameter) the low power objective lens. What is the estimated diameter of the low-power field in
millimeters?
3. Convert this distance to microns (1mm equal 1000μ). Calculate the diameter of the low-power
field in microns.
4. Repeat the above procedure to determine the diameter of the x10 objective lens in microns.

5. Since the diameter of the high power field (x400) is less than 1mm, it may be necessary to
determine the diameter of this field by calculation in terms of the measured diameter of the low
power field.
6. Calculate the diameter of the high power field in a) millimeters? b) Microns?

Activity 2
1. Remove one strand of hair from your head. Fix it to a microscope slide using a coverslip.

2. Position the slide on the stage. Find its width in millimeters and microns starting with the low
power objective lens and moving up to x10 then x40 objective lens

Observation/Calculations
(In this case it would be the answering of the questions about the diameter of the 3 different objective lenses in microns and
millimetres). Show all calculations; it would be great if you used words to demonstrate how you arrived at your answers.

Discussion:
Define microscopy. Add …why the need for a microscope.
Discuss along with other points advantages of this graph paper method to calibrate the microscope,
limitations of using the graph paper method-research use of ocular micrometer etc.
Remember you are also to discuss possible errors and limitations and their effect on the outcome
etc.

Conclusion: (Always conclude from your aim!).