Microscopes, Functions, Handling, Maintenance

Microscopy Workshop at Kymore Plant

India, 25 29 May 2009

Bruno Misteli

2009 Holcim/Switzerland

Parts of the Microscope

Trinocular tube
Eyepieces
Analyzer for transmitted and reflected light
Daylight filter
Field of view diaphragm for reflected light
Aperture diaphragm for reflected light
Lambda or gypsum plate
Revolving nosepiece
Objectives
Rotating stage
Swing out condenser
Aperture diaphragm
Stage drive or focusing knob
Polarizer for transmitted light
Voltage control
Field of view diaphragm for transmitted light
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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

The Field of View Diaphragm

The field of view diaphragm limits, as is

The field of view diaphragm should be

indicated by its name, the supply of light to

the area to be investigated.

opened as little as necessary, but wide

enough to make it disappear out of view.

The field of view diaphragm should be

A minimised opening limits the amount of

centred at a low opening and focused by

moving it up or down until its boundaries are
well defined.

diffuse light in the system and improves the

image contrast.

2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

The Aperture Diaphragm

The aperture diaphragm is used to adjust

Closing the aperture diaphragm decreases

the brightness of the picture.

the amount of light and increases the depth

of field and thus reveals internal structures,
here observed in polarised transmitted light
with a -plate.

The change of voltage supply to the light

source is in general not recommended to

adjust the brightness, as the colours change
when using different voltages.

2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Aperture Diaphragm Decentration (in reflected light)

Centred diaphragm

Same area observed with a decentred diaphragm

Some microscopes are equipped with an aperture diaphragm decentration device. The use

of this device is in general limited to reflected light applications. It can be used to amplify
topographic effects. This device is very useful to differentiate between hard and soft phases.
A similar effect can also be achieved when using microscopes without this device. An
obstacle, like a partly, just slightly inserted lambda plate into the light path can have a similar
effect. The identification of soft free lime crystals (left side of the centre) is here much easier
possible.
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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Khler Illumination

a) Field diaphragm not focused, not

b) Field diaphragm focused, but not centred

centred

d) Field diaphragm diameter = object field

c) Field diaphragm focused and centred,

diameter Khler illumination

but diameter too small

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Uniform Illumination by Lamp Adjustments

a) Direct filament image

b) Direct filament image

c) Reflected and direct

but decentred

in the right position

filament image in the

right position

2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Darkfield and Brightfield Illumination

Darkfield illumination

Brightfield illumination
(same area as left side)

Darkfield illumination is the classical stereo microscope illumination, which in general does not

require sample preparation efforts for observations. Only diffuse reflected light reaches the
eye. The colours are identical to the ones observed by the naked eye.
Brightfield illumination is the classical reflected light microscope illumination requiring a co-

axial light path. Etching effects (here HF-etching) become visible.

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Darkfield Illumination, External Light Source

When the microscope is equipped with a low magnifying objective

such as 4x or 2.5x with a long working distance, similar observations
to stereo-microscopes are possible. An external light source is
required for such observations. Special care is, however, required not
to spill the sample onto the optics underneath.
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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Microscope Handling
1.

Start your investigations always with a low magnification objective.

2.

Adjust the eyepiece distance until the pictures within both eyes unite
to one single picture.

3.

Adjust the crosshair in the eyepiece for the right eye until it is in
focus.

4.

Focus an object on the stage with the right eye by adjusting the stage
position. Start with a low position of the stage and move it slowly
towards the objective lens until a sharp picture is achieved.

5.

Leave the stage in the same position and adjust the left eyepiece until
the object is in focus.

6.

The object and the crosshair should now be perfectly in focus, if not,
start again at point 2.

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

What To Do in Case of Blurred Pictures?

Blurred pictures are often caused by dust or smears on optical parts.

Clean eyepieces with an optical cleaning liquid and a lint free tissue.

Never use solvents, as the lens fitting material might be attacked. If no

commercial cleaning liquid is at hand, use distilled water with a trace of
liquid soap.

Check the light path for any blockages, like inadvertently half inserted plate, etc.

If the picture is not in focus on one side, make sure that the surface of
the object under investigation is perfectly perpendicular to the optical
axis of the microscope.

Use an air brush in case of dust on the polished section.

Re-impregnate and re-prepare highly porous samples causing excessive

diffuse reflections.

If an objective appears to be smeared, remove the objective from its

socket and check it by looking through from the back side. Clean it with
an optical cleaning fluid and check its alignment (see next slide).

2009 Holcim/Switzerland

Microscopy Workshop 2009

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17.05.2009

Alignment of Objectives
Check whether a particle under the cross hair

remains in place while the stage is rotated.

In case this particle describes a circle, adjust the

objective by means of its two objective adjustment

screws in the revolving nose piece.

1
Stop rotating the table when the particle is in

position 2.
Move the particle by means of the objective

adjustment screws to point 3.

Move the particle from position 3 to position 1 by

means of the x-y object stage or by manual

movement.
Check again by rotating the table and repeat the

procedure if necessary.

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

How To Maintain a Good Optical Quality

Helpful articles, when fighting dust or smears on optical parts

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009

Treat Your Microscope Carefully

Cover up your microscope when not in use, to protect it from dust.

Never expose your microscope to acid vapors. When working with

hydrofluoric vapor etches, make sure your sample is clean and
sufficiently blown with a hairdryer before you take it to the
microscope.

When focusing problems occur at higher magnifications, go back to

lower magnifications to find the focus again. You can thus avoid to
damage optics or samples by crashing into them.

If necessary adjust the vertical position of all objectives to the same

focus plane by inserting special washer plates.

Do not use oil to lubricate stage drive. Your stage might want to
move down by its own weight. Use silicon grease if necessary,
which does no harm to plastic parts.

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2009 Holcim/Switzerland

Microscopy Workshop 2009

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Lets Practice!

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2009 Holcim/Switzerland

Microscopy Workshop 2009

17.05.2009