Sample Preparation for different Techniques

Submitted To: Dr. Nadeem Badani

Submitted By: M. Ishtiaq Anjum
(2018-UET/MS-PHY-15)
Introduction:
Characterization of materials in the solid state, often loosely referred to as materials
characterization, can be a vast and diverse field encompassing many techniques. In the last few
decades, revolutionary changes in electronic instrumentation have increased the use of highly
effective automated instruments for obtaining analytical information on the composition,
chemistry, surface, and internal structures of solids at micrometer and nanometer scales. These
techniques are based on various underlying principles and cannot be put under one discipline or
umbrella.

The two common categories of materials characterization are microscopy and spectroscopy.
Microscopy implies obtaining magnified images to study the morphology, structure, and shape of
various features, including grains, phases, embedded phases, embedded particles, and so on.
Spectroscopy implies investigation of chemical composition and chemistry of the solid.

Within each category, different techniques may have their own restrictions, requirements, and
concerns. As the analytical instruments become more sophisticated, robust, and user friendly,
some stringency of sample specifications can be relaxed, but those fundamental to the analytical
process remain. In this assignment we a brief introduction to those sample preparation concerns
that every user should be aware of are described in detail.

1. Sample Preparation for XRD Technique:

Proper sample preparation is one of the most important requirements in the analysis of powder
samples by X-Ray diffraction. This statement is especially true for soils and clays that contain
finely divided colloids, which are poor reflectors of x-rays, as well as other types of materials
such as iron oxide coatings and organic materials that make characterization by XRD more
difficult. Sample preparation includes not only the right sample treatments to remove undesirable
substances, but also appropriate techniques to obtain desirable particle size, orientation,
thickness, etc.

Analysis of powders by XRD requires that they be extremely fine grained to achieve good
signal-to noise ratio (and avoid fluctuation in intensity), avoid spottiness and minimize preferred
orientation. Reduction of powders to fine particles also ensures enough particle participation in
the diffraction process. The recommended size range is around 1-5µm, especially if
quantification of various phases is desired. For routine qualitative evaluation of mineral
components, the samples are usually ground to pass through a 325 mesh sieve (45 µm). Grinding
is accomplished either through hand grinding or in a mechanical grinder. The effects of
excessive grinding include lattice distortion and possible formation of an amorphous layer
(Beilby layer) outside the grains.

Solid Sample Preparation

Solid samples are prepared for X-ray diffraction by grinding, which can be accomplished by
several different methods, depending on the sample matrix, the size of the sample, and/or
quantity of prepared material needed.

2. Sample Preparation for UV-Visible Spectroscopy:

UV-visible spectroscopy is used primarily to measure liquids or solutions. This mode is simpler
and allows more accurate quantitative analysis than do reflectance measurements on solids. With
this technique, a cell must contain the liquid or solution in the spectrophotometer sample area.

For determinations using UV or visible spectrophotometry, the specimen generally is dissolved

in a solvent. Unless otherwise directed in the monograph, analysts make determinations at room
temperature using a path length of 1 cm. Many solvents are suitable for these ranges, including
water, alcohols, lower hydrocarbons, ethers, and dilute solutions of strong acids and alkalis.
Precautions should be taken to use solvents that are free from contaminants that absorb in the
spectral region under examination. For the solvent, analysts typically should use water-free
methanol or alcohol or alcohol denatured by the addition of methanol but without benzene or
other interfering impurities. Solvents of special spectrophotometric quality, guaranteed to be free
from contaminants, are available commercially from several sources. Some other analytical
reagent-grade organic solvents may contain traces of impurities that absorb strongly in the UV
region. New lots of these solvents should be checked for their transparency, and analysts should
take care to use the same lot of solvent for preparation of the test solution, the standard solution,
and the blank. The best practice is to use solvents that have NLT 40% transmittance (39.9%T =
0.399A) at the wavelength of interest.
3. Sample Preparation for TGA:
Although solid samples may be nominally of the same chemical composition, there may be
considerable differences in their behavior on heating. These differences arise from structural
differences in the solid, such as the defect content, the porosity and the surface properties, which
are dependent on the way in which the sample is prepared and treated after preparation. For
example, very different behavior will generally be observed for single crystals compared to
finely ground powders of the same compound. In addition to the influence of defects on
reactivity, the thermal properties of powders differ markedly from those of the bulk material.
As the amount of sample used increases, several problems may arise. The temperature of the
sample becomes non-uniform through slow heat transfer and through either self-heating or self-
cooling as reaction occurs. Exchange of gas with the surrounding atmosphere is also decreased.
These factors may lead to irreproducibility. Even when the sample material is inhomogeneous
and hence a larger sample becomes desirable (e.g. coal and mineral samples), the sample mass
should be kept to a minimum and replicates examined for reproducibility if necessary. Small
sample masses also protect the apparatus in the event of explosion or deflagration. The sample
should be powdered where possible and spread thinly and uniformly in the container.

4. Sample Preparation for SEM:

A specimen to be analyzed by electron microscopy has to be dry which most biological samples
are not. As dehydration might lead to structural changes, the specimens are first fixed to preserve
their structural features. Fixation is the first step and can be achieved using chemical methods
such as fixation with glutaraldehyde or physical methods such as cryofixation in liquid nitrogen.
The fixed specimens are then dehydrated usually by exposing them to an increasing gradient of
ethanol (up to 100%). The specimens are then dried using critical point method. The dried
specimens are then coated with a conducting material usually gold to make the surface
conducting and cause it emit more secondary electrons.
 5. Sample Preparation for TEM:

Preparation of specimens is the most tedious step in TEM. To make the material electronically
transparent material thickness is limited. Specimens have to be prepared with thickness of ~ 100
nm. For higher atomic weight material, the specimen has to be thinner. TEM sample preparation
depends on type of materials used.

For bulk material TEM specimen preparation is done in two steps:

 Pre-thinning : thinning to 0.1 mm thickness

 Final-thinning: thinning to 100 nm thickness

Pre-thinning: In pre-thinning stage specimen less than 1mm thick is prepared by mechanical
cutting (with a diamond saw). Then 3-mm-diameter disc is cut using punch before further
reduction of thickness. Grinding is most commonly used to reduce the thickness of metal and
ceramic specimens

Final thinning: Final thinning is mainly done by electrolytic thinning & ion milling, which
create a dimpled area on pre-thinned specimens that have regions of electron transparency as
shown in figure below. In the electrolytic thinning, metal specimen is made the anode in an
electrolytic cell. On passing current the metal is gradually dissolved and deposited on the
cathode. Finally tiny holes appear, the edge of which are suitable for electron transparency. In
ion milling method samples are bombarded with beam of energetic ions to reduce the thickness
by knocking atoms out of the specimen. This method can also be used for ceramics and other
non-conducting materials. For polymeric and biological specimens ultramicrotomy method is
used where specimen is cut into thin sections by cutting tool such as glass knife or diamond
knife.
 Thinning of bulk specimen for TEM analysis

For powder samples, such as catalysts, the powder size is reduced by grinding; to a very fine size
till the particle thickness is small enough to allow electron transmission. These fine particles are
then suspended in volatile solvent such as isopropanol. A drop of this particle suspension is
placed on thin carbon foil supported by a conventional microscope grid. On evaporation of
solvents the powder particles are ready for the observation. Alternatively the powders can be
embedded in some suitable matrix such as epoxy resin or metals, from which a flat sheet of 3
mm diameter is cut out to study by conventional methods.

6. Sample Preparation for FTIR Spectroscopy:

IR spectra can be measured using liquid, solid, or gaseous samples that are placed in the beam of
infrared light.

Liquid Sample: A drop of a liquid can be placed as a thin film between two salt plates made of
NaCl or KBr, which are transparent to infrared light at most important frequencies.

Solid Sample: A solid can be ground with KBr and pressed into a disk that is placed in the light
beam. Alternatively, a solid sample can be ground into a pasty mull with paraffin oil. As with a
liquid, the mull is placed between two salt plates. Solids can also be dissolved in common
solvents such as CH2Cl2, CCl4, or CS2 that do not have absorptions in the areas of interest.

Gaseous Sample: Gases are placed in a longer cell with polished salt windows. These gas cells
often contain mirrors that reflect the beam through the cell several times for stronger absorption.