“Fostering and empowering society in building and serving the nation”

Name:______________________ Class:____________ Date:______

Formative Assessment : Microscope Workshop - Calibrating an eye piece graticule

Eye piece graticules are useful for measuring the actual size of a
specimen. They are etched onto the eye piece and do not change in
size
Each division on the eye piece graticule will therefore represent different
values depending on the magnification being used.
Each division has to be calculated using a STAGE MICROMETER – this
does change size as it is place on the stage!
This is what a typical stage micrometer looks like

Worked example:
Example 1 – Low power objective used (x4)

The stage micrometer has 100 divisions, each are 0.1mm apart.

Suppose the full length of the eye piece graticule scale (which has 100 divisions) covered 25
divisions of the stage micrometer.

Then the full length of the eye piece scale is therefore equivalent to (25 x 0.1mm) = 2.5mm or 2500
µm long.

The eyepiece graticule has 100 divisions, so each division will measure 25000/100 = 25µm for this
magnification.

Each division of the eyepiece graticule (µm) = Length of stage micrometer (µm)

Number of eye piece divisions

Imagine eyes
(eye piece
graticule)
looking up at
the stage
(micrometer)
!
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Using the tutorial on https://www.microscopyu.com/tutorials/reticlecalibration

X and Y translation
Switch to specimen
sliders allow you to
once you have
position the
calibrated all the
eyepiece
objective powers
horizontally or
vertically in order
line it up with the
Change to fine focus
stage micrometer
to make the
movements smaller.
Start on coarse. You can change the
colour of the scale to
contrast against the
Choose a specimen Choose an objective background when
to measure after you power from the drop viewing the
have calibrated the down menu specimens
eyepiece.
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TASK 1
Choose each of the objective magnifications, calculate what each small division on the eye
piece measures at:
Magnification Number of divisions Total length What each eye
of eye piece graticule measured using piece graticule
the stage division represents
micrometer (µm) (µm)
Remember each
division = 0.1mm or
100 µm
X10

X20

X40

X60

X100

TASK 2 Measuring the specimens

Specimen Actual size (µm)

Arrowroot starch
grain at x20

Human
erythrocyte (RBC)
at x40
Pollen grain
At x60
Zygnema green
algae at x100
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Task 3 Finding Magnification

a) (i) The actual length of the cell between X and Y in Fig. 4.1 is 2 μm.

Calculate the magnification of the electron micrograph.

Show your working and give your answer to the nearest whole number.

magnification × ..........................[3]

[Total mark :23]

End of Assessment