25/07/2017 Measuring with a Microscope

MEASURING WITH A MICROSCOPE

Microscope Measuring
When we measure everyday items most of us will grab a ruler in order to make a measurement. When
using a microscope, generally magnification is higher and a ruler will not work unless you are using a
very low power stereo microscope. When using a compound high power microscope an eyepiece
reticle is used to make measurements. Most measurements that are made with a compound
microscope are between 0.2um to 25mm. It is hard to make measurements above 25mm becasue
most eyepieces on a microscope do not have a field of view greater than 25mm.
When you look through the eyepiece of a microscope that has a reticle installed in it the ruler (or cross-
line, grid, whatever is on the reticle) is super-imposed upon your image. This makes it easy to make
quick and accurate measurements, as long as you have calibrated your microscope.
Measuring Definitions
Stage Micrometer -a microscope slide (generally 1" x 3") that has a ruler etched on it. It is either made
of glass (for transmitted light) or metal (for reflected light). The stage micrometer is used to calibrate an
eyepiece reticle when making measurements with a microscope.
Eyepiece Reticle (or reticule) -a small piece of glass with a ruler etched into it that fits into a
microscope eyepiece. This ruler is used to make measurements of objects viewed through the
microscope. The image from the eyepiece reticle is imposed upon the image when looking through the
microscope. There are many variations of eyepiece reticles including lines, cross-lines, grids, counting
reticles, etc. Before purchasing an eyepiece reticle you would want to make sure your microscope
eyepiece accepts a reticle, and you would want to determine the diameter of the reticle that fits in your
eyepiece.
Field of View - the diameter of the circle of view that you see when looking through the microscope.
When calibrating an eyepiece reticle it is best to use a stage micrometer where most if not all of the
ruler on the stage micrometer fits into the field of view. The higher your magnification, the smaller your
field of view will be.
Detent - when turning the zoom knob on a stereo zoom microscope, detent is a click stop at each of
the objective values. Detent is helpful if you need to make measurements. The detent allows the zoom
knob to click into set objective values so you know that you are on a precise objective value. Trying to
make measurements with a stereo zoom microscope that does not have detent will result in inaccurate
measurements.
Why it is important to calibrate your microscope!
Even though you may be using two identical microscopes - let's say each with 10x eyepieces and a
40x objective, they can stil have slightly different magnification factors. Therefore, it is important to
calibrate your eyepiece reticle with a stage micrometer before making measurements with your
eyepiece reticle. This ensures that you will be making accurate measurements with your microscope.
How to: Microscope Eyepiece Reticle Calibration
With the eyepiece reticle installed and a stage micrometer under the microscope, focus so that both
scales are cleary in focus and lined up at the "0" point. You will want to calibrate for each objective that
you plan to use to make measurements.
Notice on the scale that the stage micrometer and the eyepiece micrometer (reticle) line up almost
exactly at 30 on the eyepiece micrometer, but seem to line up more closely at 60. The larger the
number on the eyepiece micrometer, the more accurate your calibration will be.
The stage micrometer is 1mm long with 100 divisions, which means that each longer division on the
stage micrometer = 0.01mm or 10um. If you count the lines on the stage micrometer where the

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25/07/2017 Measuring with a Microscope

eyepiece micrometer matched up with it at

60, it equals 400um. So we take 400um / 60
eyepiece reticle reading and we can
determine that with that specific objective
each division on the eyepiece reticle =
6.67um

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