Digital Imaging Solutions

In Practice
TEM Imaging

Dangerous Fiber - Electron microscopy and

digital image analysis providing occupational safety

Application Report: Electron microscopy and digital image analysis providing occupational safety

Dangerous fiber
Asbestos fibers may cause lung cancer. New types of nanoparticles also present a
high health risk. What kinds of fibers are contained within specific materials? How
many particles get into the air we breathe? Within an occupational safety context, the
technical office for dangerous occupational substances of a professional liability insurance association in Dsseldorf, Germany pursues these and other related questions.
A transmission electron microscope, MegaView III digital camera and the iTEM image
analysis platform are the investigators tools
Dr. Peter Stallknecht, Olympus Soft Imaging Solutions
GmBH, Germany

Avoid contact
Since the 1930s, the carcinogenic effects of asbestos have
been known. When asbestos fibers get into the lungs and
get lodged there they may cause lung cancer. Even the
tiniest amounts are dangerous. Fibers and dust may also
cause asbestosis (a kind of black lung disease caused by
asbestos). The field of occupational safety today is thus doing everything possible to prevent any sort of human contact
with asbestos. Preventing asbestos particles entering the air
we breathe is a particular concern.

in Essen, Germany is an EM 420 by Philips. Our investigative department is equipped with a scanning unit (STEM) and
an EDX analyzer. The use of the TEM is advisable when,
for example, the fibers in the material being investigated
are very thin (in liquids usually only 30 80 nm in diameter).
This makes them impossible to be seen while using a SEM.
If the EDX spectra of the asbestos and other types of fiber
in the specimen are almost completely identical, the TEM
is required to detect the asbestos in conjunction with
electron diffraction (see below).
2

Tracking down suspicious fibers: the side-mounted MegaView III digital

camera transmits TEM images and diffraction images directly to the PC.
The iTEM image analysis platform controls both camera and TEM and
provides convenient processing, evaluation and archiving of all images
and data at the PC. This greatly simplifies definite identification of fibers.
When asbestos fibers get into the lungs and get lodged there they may
cause lung cancer.

SEM, TEM, EDX and more

When investigating materials that may contain asbestos,
the standard approach is via Scanning Electron Microscopy
(SEM). When determining the shape and chemical composition of suspicious fibers, an EDX unit is used in conjunction
with Scanning Electron Microscopy (SEM) methods. The
EDX unit and SEM make energy dispersive X-ray microanalysis possible. In some cases, this method is not sufficient
for distinguishing various types of fibers. Then a Transmission Electron Microscope (TEM) is required due to its
greater magnification and resolution. The TEM that we use

Recording and archiving digitally

We use a MegaView III for digital acquisition of the TEM
images. This CCD camera is side-mounted on the wideangle port of the TEM column. The camera has a prism
that is moved into the beam path of the microscope and
intercepts the electrons in the intermediate image plane.
The software for reading out the image data and for operating the camera and microscope is an integrated part of the
iTEM image-analysis platform and includes an autofocus
routine. The camera and image analysis platform are by
Olympus Soft Imaging Solutions. Images are immediately available in digital form and thus require no physical
developing in a photo lab as in the past. Before acquiring,

Application Report: Electron microscopy and digital image analysis providing occupational safety

acquisition parameters can be optimized on the PC screen

via live image. During acquisition, images are automatically
calibrated. Once acquired, image quality can be checked
immediately and archived in the well structured iTEM database along with all acquisition parameters. If necessary,
evaluation data is saved there as well.
3

Air samples and most liquids are directly impacted onto

the capillary pore filter. The filter is then vapor-coated with
carbon and removed. The asbestos fibers are thus located
on a layer of carbon for the TEM acquisitions.

Example: talcum powder

Talcum powder is often used in industrial fields: for lubrication or as a parting compound or as a carrier medium for
other substances. In day-to-day life we also run into talcum,
eg, in cosmetics and medical products. Talcum consists
of the silicate mineral talc. Depending on the source of the
talc, talcum powder may contain traces of asbestos sometimes as much as several percent. These asbestos fibers
may get into the air and from there into the human body.
The investigations we conduct are meant to ensure that no
talc containing asbestos is used.
4

Talcum powder is used in many applications industrially and in our

day-to-day lives. It consists of the silicate mineral talc and may contain
asbestos. Our analyses ensure that non-asbestos-containing talc is
used and processed. The asbestos fibers are much more distinct than
the lamellar talc crystals within the TEM images (a, b, c). This makes it
easy to distinguish them from a talc fiber (d).
Does an asbestos disease result from ones occupation? If the air at
ones place of work (left) and in the lung tissue of an employee who
contracted lung cancer (right) both contain the same type of asbestos
fibers, this is a powerful indicator that there is a causal connection. TEM
images thus assist to prove that the disease was caused at ones place
of work and is thus to be classified as an occupational disease. (for
more detail on the preparation method used, see the main text)

Solid, liquid and gaseous

Within an occupational safety context, we inspect solid,
liquid and gaseous specimens to determine potential health
hazards due to asbestos and other fibers. Objects investigated include insulation materials, dust, chemicals, drinking
water, air samples from work areas and lung tissue. All these substances require special preparation for a TEM investigation. This is the reason why we initially dissolve solid materials and dusts in liquid and ensure an even dispersion via
ultrasound in fiber-free water. This solution is then filtered
through a capillary pore filter with a pore width of 0.2 m.

Not all fibers are equal

Talc is a layer lattice silicate and has lamellar crystals. On
the TEM images these appear as plate-like, semitransparent regions (fig. 3a, b, c). The long, fibrous structures are
chrysotile fibers (white asbestos fibers). There are talc fibers
also (fig. 3d) which are significantly different than the asbestos fibers in shape, transparency and where they break.
This talc thus contains asbestos and is not suitable for the
above-mentioned industrial or day-to-day usage.

Application Report: Electron microscopy and digital image analysis providing occupational safety

Possible nanoparticle danger?

Nanotechnology is viewed as one of the key technologies

of the future. Nano particles that are just a few nm in size
can present a health hazard, however. The toner particles
shown in fig. 5 are used in laser copiers. They consist of a
carbon center with a polymer casing. When a toner cartridge is replaced improperly, particles may escape to the
surrounding environment and may be inhaled by people. As
is readily visible on the images, these tiny particles have a
strong tendency to agglomerate. Despite subjecting them
to powerful ultrasound treatment, we were not able to separate these particles.
6

Some new kinds of nanoparticles may be very dangerous when inhaled.

This ink toner used in laser copiers consists of carbon nuclei with a
polymer coating. The TEM images show how strongly the particles stick
together.

Chrysotile or crocidolite? Figure a is ambiguous. Alongside the EDX

spectrum, the diffraction image of the crystalline structure provides
a further identification indicator. This is particularly helpful when the
specimen contains other fibers whose spectra are similar to or identical
to asbestos. Fig. b shows the diffraction image of crocidolite. Chrysotile
(c) looks different.

Application Report: Electron microscopy and digital image analysis providing occupational safety

Using diffraction patterns

Acknowledgements

Alongside images and EDX spectra, diffraction patterns

can be generated via TEM. SAD (Selected Area Diffraction)
electron diffraction makes it possible to draw inferences
regarding the crystalline structure of a fiber. Diffraction
patterns of asbestos are different than those of talc and
other comparable silicate minerals. This makes it possible
to clearly distinguish between asbestos and fiber-shaped
particles which are similar in appearance and have a similar
EDX spectrum. They behave differently if they make it into
the lungs and have different consequences health-wise.
This is what makes it so important to differentiate these
fibers for those in the occupational safety field, and for
being able to assess asbestos-caused ailments resulting at
the workplace.

Text written in close collaboration with:

Dr. Gisela Binde, Department for Prevention, Dangerous
Substances Unit, Confederation of the Professional Liability Insurance Associations for Mechanical Engineering and
Metallurgy; and for Smelting and Milling [German original:
Prventionsabteilung, Fachstelle Gefhrliche Arbeitsstoffe, Verwaltungsgemeinschaft Maschinenbau- und
Metall-Berufsgenossenschaft und Htten- und WalzwerksBerufsgenossenschaft], Hoffnungstrasse 2, D-45127 Essen,
Germany
Image source: Dr. Gisela Binde, Department for Prevention,
Hoffnungstrasse 2, D-45127 Essen, Germany
All rights including translation are reserved. Any and all forms of reproduction must be
authorized by Olympus Soft Imaging Solutions. We are unable to assume any liability
for unrequested consignments received.

There are various types of asbestos. Some occur naturally such

as amphibole-silicate minerals with a fibrous structure (including
blue and brown asbestos) as well as chrysotile (white asbestos) a
sheet silicate mineral with a serpentine fibrous structure. Due to its
chemical stability, heat resistance and high electrical and thermal
insulation capacity, asbestos was popular up through the 1970s. It
was used a lot for construction despite the fact that its carcinogenic effects have been known since the 1930s. It has been used
for heat insulation, for fire resistance in construction, as a sealant
material and as insulation and in brake pads. Asbestos may also occur naturally or as a production-related impurity in other materials. It
also occurs in certain chemicals due to production processes.
Asbestos is dangerous when fibers or dust thereof are released

and inhaled. The fibers of natural asbestos are so dangerous

because they cannot be broken down biologically or at best,
very slowly. This means they cause continuous irritation to lung
tissue once inhaled. The critical dimension for fibers inhaled
into the lung are longer than 5 m, thinner than 3 m and their
length/diameter ratio is greater than 3:1. When asbestos fibers
get into the lungs and get stuck there, they can cause lung
cancer and asbestosis (a kind of black lung disease caused
by asbestos). Asbestos fibers also cause the rarely curable
mesothelioma (malignant tumor of mesothelial tissue): the fibers
penetrate the outer layer of the lungs and cause irritation to the
pleura which ultimately results in cancer. Technical usage or
processing of asbestos causes asbestos dust and microfine fibers may become separated. This makes protecting employees
from any exposure to asbestos a job for occupational safety.

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Application Report asbestos #1, 09-2007

Asbestos a dangerous matter