Optical Microscope

Contents

• Working Principle
• Sample Preparation for Optical Microscope
• Evaluation of Asbestos/fibre material in Talc

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Principle

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Sample Preparation for Optical Microscope

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 The optical properties of minerals refer to their behavior in the presence of light and how they interact with light when
observed using various optical techniques. These properties include transparency/opacity, color, luster, refractive index
(RI), pleochroism, birefringence, dispersion, extinction, and crystallography.
 Color: The color of a mineral can be a useful diagnostic tool. However, it should be noted that color can vary greatly
depending on impurities, and so it is not always a reliable indicator of a mineral’s identity.
 Luster: Luster refers to the way that a mineral reflects light. Minerals can be metallic, glassy, pearly, or dull, and each
type of luster can be used to help identify a mineral.
 Transparency: Some minerals are transparent, while others are opaque. Minerals that are transparent can be further
categorized as either colorless, colored, or pleochroic (displaying different colors when viewed from different angles).
 Refractive index: The refractive index of a mineral is a measure of how much light is bent as it passes through the
mineral. This property can be used to identify a mineral by measuring the angle at which light is refracted.
 Birefringence: Birefringence refers to the property of a mineral that causes light to split into two rays as it passes
through the mineral. This property is particularly useful for identifying minerals in thin sections under a microscope.
 Dispersion: Dispersion refers to the way that different colors of light are refracted at different angles by a mineral. This
property is particularly useful for identifying gems such as diamonds.
 Pleochroism: Pleochroism refers to the property of a mineral that causes it to display different colors when viewed
from different angles.
 Fluorescence: Some minerals exhibit fluorescence, meaning that they emit light when exposed to ultraviolet light. This
property can be used to help identify minerals in certain settings.
 Overall, optical properties are an important diagnostic tool for identifying minerals. By understanding these properties
and how they relate to each other, mineralogists can determine the identity of a mineral with a high degree of accuracy.
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• Optical microscopy, also known as light microscopy, is a widely used technique in the
field of mineralogy for the identification and characterization of minerals. It involves
the use of a microscope that utilizes visible light to magnify and analyze mineral
samples. Here are some key points about optical microscopy in mineralogy
• Principle: Optical microscopy is based on the interaction of light with minerals.
When light passes through a mineral sample, it can be absorbed, transmitted, or
reflected depending on the mineral’s optical properties, such as color, transparency,
and refractive index. By observing how light interacts with a mineral under a
microscope, valuable information about its physical and optical properties can be
obtained.
• Equipment: Optical microscopy requires a specialized microscope equipped with
various components, including a light source, lenses, a stage for holding the mineral
sample, and eyepieces or a camera for viewing and capturing images. Polarizing
microscopes, which use polarized light, are commonly used in mineralogy for
studying the optical properties of minerals.
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• Sample Preparation: Mineral samples for optical microscopy are typically thin sections or polished
thin mounts, which are prepared by cutting a thin slice of a mineral specimen and mounting it on a
glass slide. Thin sections are commonly used for studying the mineralogy of rocks, while polished
thin mounts are used for analyzing individual mineral grains.
• Techniques: Optical microscopy techniques used in mineralogy include transmitted light
microscopy, which involves passing light through a thin section or thin mount to observe the
mineral’s internal features, and polarized light microscopy, which involves the use of polarized light
to study the mineral’s optical properties, such as birefringence, extinction, and pleochroism. Other
techniques, such as reflected light microscopy and fluorescence microscopy, may also be used for
specific purposes in mineral identification and characterization.
• Mineral Identification: Optical microscopy is a powerful tool for mineral identification based on
their physical and optical properties. By observing a mineral’s color, transparency, crystal shape,
cleavage, and other features under a microscope, and by using techniques such as polarization and
interference, mineralogists can identify minerals and differentiate between different mineral species.
• Limitations: Optical microscopy has some limitations in mineralogy. It may not be suitable for
identifying minerals with similar physical and optical properties, or minerals that are very small or
opaque. In such cases, other techniques such as X-ray diffraction, electron microscopy, or
spectroscopy may be required for more accurate mineral identification and characterization.
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 Why use the microscope ?
• Microscopes are used in mineralogy for a variety of reasons:
• Mineral Identification: Microscopes are used to observe the physical and optical properties of minerals, such as color, transparency,
crystal shape, cleavage, and other features, which are essential for their identification. By examining mineral samples under a
microscope, mineralogists can gather critical information that helps them identify different mineral species and distinguish between
similar minerals.
• Mineral Characterization: Microscopy allows for the detailed characterization of minerals, including their crystal structure, texture,
and inclusions. This information provides insights into the formation and history of minerals, which can be important for
understanding their properties and applications.
• Mineralogical Research: Microscopy is used in mineralogical research to study the optical, chemical, and physical properties of
minerals, as well as their relationships with other minerals and rocks. Microscopic analysis can provide valuable data for
understanding mineral occurrences, mineralogical processes, and geological history.
• Mineral Processing: Microscopy is used in the field of mineral processing to analyze and optimize the beneficiation of ores and
minerals. By examining mineral samples under a microscope, mineral processing experts can assess the mineral liberation, mineral
associations, and mineralogical characteristics of ores, which can help in developing effective mineral processing strategies.
• Geological Mapping: Microscopy can be used in geological mapping and mineral exploration to identify and map minerals in rocks
and ores. This information can be used to understand the distribution, composition, and economic potential of mineral deposits in a
given area.
• Education and Teaching: Microscopes are widely used in educational settings to teach students about mineralogy and geology. By
using microscopes, students can observe and identify minerals, and learn about their properties, occurrences, and uses.
• In summary, microscopes are essential tools in mineralogy for mineral identification, characterization, research, mineral processing,
geological mapping, and education. They allow for detailed observation and analysis of minerals, providing valuable insights into their
properties, occurrences, and applications. 9
 Minerals and propagation of light
• The propagation of light through minerals is a fascinating topic in mineralogy and is closely related to the optical
properties of minerals. When light passes through a mineral, it may undergo various interactions, such as
absorption, reflection, refraction, and polarization, which can provide important information about the mineral’s
composition, structure, and properties. Here are some key points related to the propagation of light in minerals:
• Transparency and Opacity: Minerals can be transparent, translucent, or opaque to light, depending on their
chemical composition and internal structure. Transparent minerals allow light to pass through with little or no
scattering, while translucent minerals scatter light to some extent, and opaque minerals do not allow light to pass
through at all.
• Absorption: Some minerals have selective absorption of certain wavelengths of light due to the presence of specific
chemical elements or compounds. This results in the mineral appearing colored when viewed under a microscope or
with the naked eye. The absorption spectrum of a mineral can provide information about its chemical composition.
• Refraction: Refraction is the bending of light as it passes from one medium to another with a different refractive
index. Minerals with different crystal structures and chemical compositions can exhibit different refractive indices,
which can be determined using a refractometer. The refractive index is an important optical property used in
mineral identification.
• Polarization: Light passing through certain minerals can become polarized, meaning the light waves oscillate in a
particular direction. This property can be observed using a polarizing microscope, which allows for the examination
of minerals in cross-polarized light. Polarized light microscopy is a powerful technique used in mineral identification
and characterization.
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 Minerals and propagation of light

• Pleochroism: Some minerals exhibit pleochroism, which means they show different colors when viewed
from different angles under polarized light. This property is caused by the preferential absorption of light in
different directions due to the mineral’s crystal structure and can be used as a diagnostic tool in mineral
identification.
• Birefringence: Birefringence, also known as double refraction, is the property of certain minerals to split
light into two rays with different refractive indices. This can be observed using a polarizing microscope, and
the amount of birefringence can provide information about the mineral’s crystal structure and composition.
• Optical Sign: The optical sign of a mineral refers to the direction in which the mineral’s refractive indices
are oriented with respect to its crystallographic axes. The optical sign can be determined using a polarizing
microscope and is an important characteristic used in mineral identification.
• The study of how light interacts with minerals and how it propagates through them is crucial in mineralogy,
as it provides important information about the mineral’s composition, structure, and properties. Optical
properties of minerals, such as absorption, refraction, polarization, pleochroism, birefringence, and optical
sign, are used in mineral identification, characterization, and research. Microscopic techniques, such as
polarizing microscopy, are widely used to study the propagation of light through minerals and reveal
important details about their optical properties.
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Thin section
• A thin section refers to a thin slice of a rock or mineral that is mounted on a glass slide and
ground down to a thickness of typically 30 micrometers (0.03 mm) using specialized
equipment. Thin sections are used in petrology, a branch of geology that studies rocks and
minerals under a microscope to determine their mineral composition, texture, and other
important characteristics.
• Thin sections are created by cutting a small piece of rock or mineral into a thin slab, which
is then affixed to a glass slide using an adhesive. The slab is then ground down to the
desired thickness using a series of abrasive materials, such as silicon carbide powder, to
achieve a smooth and even surface. The resulting thin section is then polished to improve
transparency and clarity, and may be stained with dyes or chemicals to enhance certain
features or properties.
• Thin sections are commonly examined under a polarizing microscope, also known as a
petrographic microscope, which is equipped with polarizers and analyzers that allow for
the study of the rock or mineral’s optical properties, such as birefringence, pleochroism,
and extinction angles. By analyzing the minerals and their arrangement in the thin section,
geologists can identify the rock type, determine the mineral composition, and interpret the
rock’s history, such as its formation and deformation processes.
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• Thin sections are widely used in various fields of geology, including igneous
petrology, sedimentary petrology, metamorphic petrology, economic geology, and
environmental geology. They are essential tools for studying rocks and minerals at a
microscopic level and provide valuable insights into their origin, evolution, and
properties. Thin sections are also commonly used in education and research, as they
allow for detailed examination and analysis of rocks and minerals, contributing to
our understanding of Earth’s geology and its history.

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Properties of Light
• Wave-like nature: Light exhibits wave-like properties, such as wavelength, frequency, and amplitude. It can be
described as an electromagnetic wave that travels through a medium or vacuum.
• Particle-like nature: Light also behaves as a stream of particles called photons, which carry energy and momentum.
• Speed: Light travels at a constant speed of about 299,792 kilometers per second (km/s) in a vacuum, which is the
fastest known speed in the universe.
• Electromagnetic spectrum: Light exists in a range of wavelengths and frequencies, which together form the
electromagnetic spectrum. This spectrum includes different types of light, such as visible light, ultraviolet (UV)
light, infrared (IR) light, X-rays, and gamma rays, each with its own unique properties and uses

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Plane Polarized Light (PPL)
• Polarization: Light waves can be polarized, which means that their oscillations occur in a
single plane, as opposed to in all directions. Polarized light has a specific orientation of its
electric field vector.
• Polarizers: PPL is created by passing unpolarized light through a polarizer, which is a filter that
transmits only the light waves oscillating in a specific plane while blocking those oscillating in
other planes.
• Properties: PPL has properties such as direction, intensity, and color that can be used to study
and analyze various materials, such as minerals and crystals, under a polarizing microscope.

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XPL (Crossed Polarizers)
• Technique: XPL is a technique used in polarized light microscopy, where two polarizers are
crossed, meaning their polarization planes are perpendicular to each other.
• Interference: When a thin section of a mineral or a crystal is placed between crossed polarizers,
it can create interference patterns known as interference colors or birefringence, which provide
information about the mineral’s optical properties, such as refractive index and crystal structure.
• Identifying minerals: XPL is commonly used in mineralogy to identify and characterize
minerals based on their unique interference patterns and birefringence colors, which can help in
determining the mineral’s composition, crystal structure, and other properties.

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Passage of Light
• Reflection is a process in which light, or other forms of electromagnetic radiation, bounces off a surface and
returns back into the same medium from which it originated, without changing its frequency or wavelength. This
phenomenon occurs when light encounters a boundary between two media with different refractive indices or
optical densities.
• Key points about reflection:
• Angle of incidence and angle of reflection: The angle at which light strikes a surface is called the angle of
incidence, and the angle at which it is reflected is called the angle of reflection. According to the law of reflection,
the angle of incidence is equal to the angle of reflection, and the incident ray, the reflected ray, and the normal (a
line perpendicular to the surface) all lie in the same plane.
• Specular vs. diffuse reflection: Reflection can be either specular or diffuse. Specular reflection occurs when light
reflects off a smooth surface, such as a mirror, and the reflected rays maintain their original direction and form a
clear reflection. Diffuse reflection occurs when light reflects off a rough or irregular surface, such as paper or a
matte surface, and the reflected rays scatter in different directions, resulting in a less clear reflection.
• Applications of reflection: Reflection is used in many everyday applications, such as mirrors, reflective surfaces
on vehicles and road signs for visibility, optical devices like telescopes and microscopes, and in photography and
art for creating visual effects.
• Law of reflection: The law of reflection states that the angle of incidence is equal to the angle of reflection, and the
incident ray, the reflected ray, and the normal all lie in the same plane. This law is fundamental in understanding
the behavior of light when it encounters a reflective surface.
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Passage of Light

• In summary, reflection is the process in which light or other forms of electromagnetic

radiation bounce off a surface and return back into the same medium from which it
originated, without changing its frequency or wavelength. It involves the angle of
incidence and angle of reflection, can be specular or diffuse, has many practical
applications, and follows the law of reflection.

Reflection passage of Light(Refraction)

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• The velocity of light depends on the medium through which it passes. Light is an electromagnetic
wave which interacts with electrons.The distribution of electrons are different for each material and
sometimes for different directions through a material. When light passes from one medium to
another there is a difference in velocity. Light rays apparently bend at the contact
• Angle of incidence ≠ Angle of Refraction.
Refractive Index
• The amount of refraction is related to the difference in velocity of light in each medium. Refractive
index (R.I.) for air is defined as 1
• The absolute refractive index for a mineral (n) is the refraction relative to that in air.
• depends on the atomic/crystal structure
• is different for each mineral
• is constant for a mineral
• is a diagnostic property of the mineral
• between 1.3 and 2.0
• There may be one, two or three values of R.I. depending on the atomic structure of the mineral.20
Opaque Mineral
• Opaque minerals are minerals that do not transmit light and do not allow light to pass through them. They
appear opaque or dull when viewed under a microscope or with the naked eye, as they do not have the ability
to transmit light through their structure.
• Opaque minerals are typically composed of materials that are not transparent or translucent to light due to
their physical and chemical properties. They may contain various impurities, minerals, or elements that
absorb or scatter light, preventing it from passing through.
• Some examples of opaque minerals include native metals like gold, silver, and copper, as well as sulfides
like pyrite, galena, and chalcopyrite. These minerals are commonly found in ore deposits and are often
associated with metallic ore deposits. Other opaque minerals include certain oxides, carbonates, and sulfates,
which can have metallic or non-metallic compositions.

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 Transparent mineral
• Transparent minerals are minerals that allow light to pass through them, making them appear clear or translucent when
viewed under a microscope or with the naked eye. These minerals have a crystalline structure that allows light to pass
through their lattice, allowing them to transmit light without scattering or absorbing it.
• Transparent minerals can be found in a wide range of colors and can exhibit various optical properties such as pleochroism
(change in color with orientation), birefringence (double refraction), and interference colors when viewed under a polarized
light microscope. These properties can be used to identify and differentiate transparent minerals.
• Some examples of transparent minerals include quartz, calcite, feldspar, garnet, tourmaline, and topaz. These minerals are
commonly found in rocks and minerals from various geological settings and have diverse applications in industry, jewellery,
and scientific research.

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Becke Line

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Sample Preparation for Optical Microscope

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PCM-PLM-Dispersion Staining
Go/No-Go Test for Amphibole
Fibers in Talc

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Introduction
• Objective:- The PCM-PLM Dispersion Staining Go/No-Go test is used as a screening test to determine whether
or not amphibole fibers are present in powdered samples of talc.
• A sample is examined using a polarizing phase-contrast microscope (PCM-PLM) at 400X and 160X
magnifications. Fibers with diameters greater than 0.5 to 1 micrometer may be examined for the presence of
retardation colors. If there are any visible fibers, they are examined using central stop dispersion staining
(CSDS) using the same microscope.
• A preparation of powdered sample is made in high-dispersion refractive index liquid with n = 1.605. The
observable indices of refraction of all of the regulated amphiboles are greater than 1.605, while the indices of
refraction for talc is less than n = 1.605.
• Because of this, the central-stop dispersion staining (CSDS) colors observed for amphiboles will indicate that
the particle indices are greater than the index of the liquid. Generally, this will be yellow orange to yellow, or
yellow-white. Talc, brucite and chrysotile fibers will be blue to blue-white.
• If there are no fibers with CSDS colors indicative of possible amphibole present (e.g. yellow to yellow white)
then it is unlikely that any amphibole asbestos is present.
• When fibers are visible that have CSDS colors indicating indices greater than n = 1.605, further examination in
matching liquids as well as other measurements are required to identify the amphibole.
• The index of refraction liquid n = 1.605 is not a matching liquid for talc or any amphibole and is insufficient to
identify any of the regulated minerals.
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Introduction
• The PCM-PLM Dispersion Staining Go/No-Go test is used as a screening test to determine whether or not
amphibole fibers are present in powdered samples of talc.
• A sample is examined using a polarizing phase-contrast microscope (PCM-PLM) at 400X and 160X magnifications.
Any fibers meeting the federal definition for asbestos will be visible because of the use of PCM. Fibers with
diameters greater than 0.5 to 1 micrometer may be examined for the presence of retardation colors. If there are
any visible fibers, they are examined using central stop dispersion staining (CSDS) using the same microscope.
• A preparation of powdered sample is made in high-dispersion refractive index liquid with n = 1.605. The
observable indices of refraction of all of the regulated amphiboles are greater than 1.605, while the indices of
refraction for talc is less than n = 1.605.
• Because of this, the central-stop dispersion staining (CSDS) colors observed for amphiboles will indicate that the
particle indices are greater than the index of the liquid. Generally, this will be yellow orange to yellow, or yellow-
white. Talc, brucite and chrysotile fibers will be blue to blue-white.
• If there are no fibers with CSDS colors indicative of possible amphibole present (e.g. yellow to yellow white) then
it is unlikely that any amphibole asbestos is present.
• When fibers are visible that have CSDS colors indicating indices greater than n = 1.605, further examination in
matching liquids as well as other measurements are required to identify the amphibole.
• The index of refraction liquid n = 1.605 is not a matching liquid for talc or any amphibole and is insufficient to
identify any of the regulated minerals.

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• The samples submitted for this investigation were either packed powder or loose powder. All were
apparently dry with no apparent wetness, equilibrated to the local humidity.
• For this examination, no gravimetric reduction was performed. (e.g. the samples were not subjected to
acid, heat or chemicals designed to remove unwanted substances, and examined as received.).
• Samples were prepared individually in a table-top laboratory hood, and all instruments and horizontal
surfaces were cleaned in between samples. This is a measure to prevent cross-contamination between
the samples.
At least three separate slide preparations were made and examined for each of the samples.
• Two to three drops of high dispersion index of refraction liquid are allowed to drop by gravity onto the
surface of a cleaned 1 x 3 in. glass slide.
• A clean dissecting needle is used as a sampling probe. The tip is wetted in the index of refraction liquid
on the slide and then dipped into the sample.
• A small amount of powder clings to the dissection needle. The amount is determined by the diameter of
the captured powder clump and is usually less than a 0.5 mm.
• The dissection needle with the powder is introduced into the liquid on the slide and stirred gently to
disperse the powder as evenly as possible.
• A clean, No 1 ½ glass cover slip is gently lowered onto the liquid. 32
• NOTE 1: The amount of index of refraction liquid placed on the slide in step 1 is
determined by experience such that little to no liquid extends beyond the boundary
of the cover slip.
• NOTE 2: If the preparation is cloudy, it indicates that too much powder was on the
dissection needle when the powder was sampled. Discard this preparation and
perform steps 1 to 4 using a smaller amount of powder until a suitable preparation is
made. Too much powder in the preparation will interfere with the optical tests.
These tests generally depend on particles being dispersed in the liquid sufficiently
far apart so that their images do not interfere with one another.

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Examination
• A phase contrast microscope which also has crossed polarizing elements and a first-
order red compensator is used to examine the samples.
• A slide preparation is placed on the microscope stage and examined at 160X and
400X. (The sample may also be examined at 100X if the microscope is equipped for
PCM at 100X. In this particular investigation, little use was made of the 100X
magnification because the particles were very finely divided.)
• An initial scan of the slide is made at 160X and 400X to note the presence of any
elongate particles. The morphology of any observed elongate particle is evaluated
whether it is, organic, synthetic or mineral, and if mineral, consistent with asbestiform
growth habit or other growth habit. Any observed fiber is evaluated for the presence
of birefringence observed as retardation colors.
• Any mineral fibers are examined at 160X using CSDS. If the colors observed indicate
that the fiber may be amphibole, further analysis of the sample is indicated. The
sample prep is scanned at 160X to determine if any amphibole source mineral is
present.
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SOP for Optical Microscope

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SCOPE & PURPOSE

• The purpose of this SOP is to show researchers and staff how to operate the
microscope safely in the New R&D Laboratory.
• How to cleaning the optical microscope.
• How to maintenance for optical microscope.

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Responsibility & Accountability

Responsibility:
Chemist, Senior Chemist
R&D Manager – Check the SOP

Accountability:
R&D Department Head- Compliance the SOP

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POTENTIAL HAZARDS/HEALTH EFFECTS:

• Contact with objects at high temperatures – the globe and other elements can become hot
if used for prolonged times. Ergonomic due to other factors – long periods of sitting.

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PRECAUTIONS AND NOTES
• Raise the specimen stage slowly and carefully in order to avoid contact between the
objective and wafer
• Changing the magnification gradually in order to prevent the objective hitting the
sample. Always start at 50 and increase gradually(i.e. from 50 to 100 of 50 to 1000)
• Never place a wet sample to image/analysis a sample that is wet or contains a strong
acid, solvent or base, which may damage the objectives.
• Use the Kodak lens cleaning paper and Edmund Scientific lens luster cleaning fluid for
cleaning the microscope eyepieces.

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INSTRUCTIONS:
Pre-Start check:
 Ensure that the microscope and its surrounding area is clean.
 Check plug in the microscope computer to the shared monitor.
 Check turn the brightness dial to its minimum.
 Choose the correct specimen holder and mount on the stage.
Operation: For additional information on how to perform these tasks consult the appendix and/or manual
 Turn the microscope power on at front right.
 Place the specimen on the stage plate.
 Engage the lowest magnification objective using the revolving nosepiece.
 Adjust the illumination brightness.
 Focus on the sample by adjusting the coarse and then fine adjustment knobs.
 Adjust the interpupillary distance until the left and right fields of view coincide completely and so that the two index
dots are horizontal.
 Focus on the sample looking only through the right eyepiece, and then looking only through the left eyepiece adjust
the diopter to focus on the sample.
 Engage the desired magnification objective using the revolving nosepiece.
 Focus on the sample using the coarse and then fine adjustment knobs. 40
• Adjust the illumination brightness.
• Adjust the contrast, depth of focus and resolution to your requirements by removing
the eyepiece, looking into the eyepiece sleeve and using the aperture iris diaphragm
(70-80% of objective pupil) and adjusting the aperture iris diaphragm lever.
After Use:
• Return the magnification objective to the lowest setting.
• Return the brightness dial to its minimum.
• Turn off the microscope.
• Put on the microscope cover.

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Operation Procedure

• Plug the microscope power cord in to electrical out let.

• Turn on the microscope by rotating the illumination control knob on the bottom left side of the instrument.
• Set the intensity of light to the lowest setting using illumination control knob.
• Load the wafer or device on the specimen stage and move the stage so that the device is positioned under the 5
objective. (The adjustment knobs are under the stage. The upper one is for "x" adjustment, the lower one is for
"y" adjustment.)
• Choose an appropriate objective to get the desired magnification by turning the nosepiece. Start out at low
magnification (50) and work your way up to the higher magnifications.
Note: Microscope magnification = individual objective magnification multiplied by the eyepiece magnification
(10x in this microscope).
• Adjust the eyepiece spacing to fit your eye spacing by moving the eyepieces in and out until a complete circular
image is obtained when viewing through both eyepieces.
• Fully open the aperture diaphragm of the condenser by rotating the ring to the extreme right.
• Using the sub stage condenser focusing knob, raise the condenser to the top of its excursion. Critical
illumination only: If the condenser travel is excessive, limit the travel with the thumbscrew under the sub stage
until the top lens of it is just below the stage surface (0.35mm)
• Place the specimen slide on the stage.
• Rotate the nosepiece to move the objective (40 X for dry mount and 10 X for wet mount) into working position.
• Raise the stage by rotating the coarse adjustment knob to its positive stop. Using the fine adjustment knob,
bring the specimen into sharp focus.
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PROCEDURE
• Turn on the DC power source for the lamp and adjust needle to around 10 volts.
NOTE: DO NOT operate the needle in the red.
• Load the wafer or device on the specimen stage and move the stage so that the device
is positioned under the 5 objective. (The adjustment knobs are under the stage. The
upper one is for "x" adjustment, the lower one is for "y" adjustment.)
• Choose an appropriate objective to get the desired magnification by turning the
nosepiece. Start out at low magnification (50) and work your way up to the higher
magnifications.
Note: Microscope magnification = individual objective magnification multiplied by
the eyepiece magnification (10x in this microscope).
• Adjust the eyepiece spacing to fit your eye spacing by moving the eyepieces in and
out until a complete circular image is obtained when viewing through both
eyepieces.

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• Adjust the eye tubes for inter pupillary distance and eye difference. The left eyepiece tube
is focusable to compensate for refractive differences of the eyes.
• To correctly set the eye tubes, focus on the specimen through the right eyepiece tube only.
Use the fine adjustment knob while covering the left eyepiece or closing the left eye.
• Next, focus the specimen through the left eyepiece by turning the eye tube. Cover the right
eyepiece while doing this and be sure to focus with the left eye tube only, without using the
focusing knob.
• Remove an eyepiece and view the back aperture of the objective. Close the condenser
aperture diaphragm and then, to obtain the full resolving power of the microscope, reopen
until the diaphragm leaves just disappear from view. Replace the eyepiece. The aperture
diaphragm can be adjusted to enhance contrast and/or increase the depth of focus.
• When changing to higher power objectives, the positions of the aperture diaphragm must
be reset. As magnification increases, the aperture diaphragm must be opened as required.

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Focusing
• Raise the specimen stage by turning coarse focusing control (the large knob) until
you get an image of your device. Be careful not to contact the objective with your
sample.
• Use the fine focusing control (the small knob) to get a clear image.
Illumination: Brightfield and darkfield illumination are available.
• Usually we use brightfield illumination to inspect devices. The reflector slider should
be in the H (brightfield) position.
• Darkfield illumination can be applied by changing the reflector slider from the H
(brightfield) position to the D (Darkfield). It is applied to examine scratches, fissures,
and pores, which deviate from the flatness of the surface. Such light scattering
features light up in darkfield, while the specular flat surfaces remain dark.
NOTE: Aperture and field irises must be open to avoid light losses when you are using
Darkfield illumination
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Pushrods and screws located on the side of the microscope.
• The setting screws for Aperture and Field Irises are for centering the individual
irises.
• Pushrods for Aperture and Field Irises: To the left -iris open. To the right -iris closed.
NOTE: Aperture Iris is for adjusting the image contrast. Field Iris is for changing the
size of viewing area.
• Pushrod for Diffuser: To the left -diffuser is out. To the right -diffuser is in.

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• Measurement: A ruler reticle in the left eyepiece allows direct measurement of
device features.
50× -> 20 µm/div
100× -> 10 µm/div
200× -> 5 µm/div
500× -> 2 µm/div
1000× -> 1 µm/div
• Photographs: A Polaroid camera attaches to the top of the microscope for easy
photographing of devices.
• When you finish with the microscope, LOWER the specimen stage first. Then take
Specimen out.
• Shut off the power for the lamp.

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Cleaning and Maintenance

• Clean the lower magnification objectives and optical surfaces with a lint free cloth or
lens tissue moistened (not wet) with methanol, if lack of contrast, cloudiness or poor
definition is encountered.
• Clean the front lens with a toothpick covered with a cotton tip wetted with methanol.
• Avoid excessive use of solvent for cleaning.
• Cover the microscope always with dust cover, whenever the microscope is not in use.
• Wipe the bottom of Oil immersion lens of a fast absorbing tissue paper before and
after using the lens.
• Use Xylene to clean the lens surfaces

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SPILLS/ ACCIDENT REPORTING PROCEDURE:

• Report incidents to: R&D Manager

In case of Emergency:
• Turn power off.
• Contact Technical Staff.
• If a serious injury has occurred call 000.
• Alert others if nearby.
• Evacuate to closest assembly area.

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REPAIRS & CERTIFICATION/VALIDATION SCHEDULE

• Reporting faults/concerns of equipment procedure: R&D Manager

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GENERAL CARE & MAINTENANCE:

• Always ensure the brightness is set to minimum before turning the power on the
microscope.
• Do not leave the microscope power on when unattended and at completion of use.
• Check lenses for dust and debris.
• Blow with air and clean lenses with lens wipes.

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