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Measurements Using Ocular and Stage Micrometers

This document provides instructions for measuring objects using an ocular micrometer and stage micrometer under a microscope at different magnifications. Students are asked to calibrate the ocular micrometer using the stage micrometer at 4x, 10x, 40x, and 100x magnification, then use the calibrated ocular micrometer to measure the dimensions of the letter "e" on a prepared slide at each magnification level. They are also asked to directly measure the letter "e" with a millimeter ruler to compare to the microscope measurements. Notes provide background on using an ocular micrometer as a scale by calibrating it against the known scale of a stage micrometer at different magnifications.

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132 views4 pages

Measurements Using Ocular and Stage Micrometers

This document provides instructions for measuring objects using an ocular micrometer and stage micrometer under a microscope at different magnifications. Students are asked to calibrate the ocular micrometer using the stage micrometer at 4x, 10x, 40x, and 100x magnification, then use the calibrated ocular micrometer to measure the dimensions of the letter "e" on a prepared slide at each magnification level. They are also asked to directly measure the letter "e" with a millimeter ruler to compare to the microscope measurements. Notes provide background on using an ocular micrometer as a scale by calibrating it against the known scale of a stage micrometer at different magnifications.

Uploaded by

ponderofbiotech
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© Attribution Non-Commercial (BY-NC)
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Measurements using Ocular and Stage Micrometers

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Materials

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Microscope

Ocular micrometer

Stage micrometer

Millimetre ruler

Prepared slide with letter e

Procedure

Place a stage micrometer on the microscope stage, and using the lowest magnification (4X), focus on the grid of the
stage micrometer.
Rotate the ocular micrometer by turning the appropriate eyepiece. Move the stage until you superimpose the lines of the
ocular micrometer upon those of the stage micrometer. With the lines of the two micrometers coinciding at one end of the
field, count the spaces of each micrometer to a point at which the lines of the micrometers coincide again (Figure 1).
Since each division of the stage micrometer measures 10 micrometers, and since you know how many ocular divisions
are equivalent to one stage division, you can now calculate the number of micrometers in each space of the ocular scale.
Repeat for 10X and 40X, and 100X. Record your calculations below.

Microscope #

Value for each ocular unit at 4X

Value for each ocular unit at 10X


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Value for each ocular unit at 40/45X

Value for each ocular unit at 100X

Using the stage micrometer, determine the smallest length (in microns) which can be resolved with each objective. This
is the measured limit of resolution for each lens. Compare this value to the theoretical limit of resolution calculated on the
basis of the numerical aperture of the lens and a wavelength of 450 nm (blue light).
Using the calculated values for your ocular micrometer, determine the dimensions of the letter e found on your
microscope slide, and add the dimensions to your drawing in Exercise 1.1. Use a millimetre ruler to measure the letter e
directly and compare with the calculated values obtained through the microscope.

Notes

To measure an object seen in a microscope, an ocular micrometer serves as a scale or rule. This is simply a disc of
glass upon which equally spaced divisions are etched. The rule may be divided into 50 subdivisions, or more rarely 100
subdivisions. To use the ocular micrometer, calibrate it against a fixed and known ruler, the stage micrometer. Stage
micrometers also come in varying lengths, but most are 2 mm long and subdivided into 0.01 mm (10 micrometer)
lengths. Each objective will need to be calibrated independently. To use, simply superimpose the ocular micrometer onto
the stage micrometer and note the relationship of the length of the ocular to the stage micrometer (Refer to Figure 1.7).
Note that at different magnifications, the stage micrometer changes, but the ocular micrometer is fixed in dimension. In
reality, the stage micrometer is also fixed, and what is changing is the power of the magnification of the objective.

Reference:

Cell Biology Laboratory Manual

Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College,

St. Peter, MN 56082 -- [email protected]

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