50 MICROSCOPY APPLICATIONS / Food
and Applications. Cambridge: Cambridge University
Press.
Steinbrecht RA and Zierold K (1987) Cryotechniques in
Biological Electron Microscopy. Berlin: Springer.
Food
R M Twyman, University of York, York, UK
& 2005, Elsevier Ltd. All Rights Reserved.
This article is a revision of the previous-edition article by M Kaláb
and S S Miller, pp. 3210–3218, & 1995, Elsevier Ltd.
Introduction
Most foods are of biological origin, and processing
changes their nature to varying degrees. Transitions
such as grain-flour-bread, muscle-meat-sala-
mi, and milk-curd-cheese may be used as exam-
ples of foods that undergo these changes. All foods
may be categorized according to specific structural
characteristics. These include liquid foods that are in
the form of suspensions and emulsions (beverages
such as milk, fruit and vegetable juices, dressings,
toppings, etc.), foamed foods (whipped cream,
marshmallows), viscous foods with a low fat con-
tent (yogurts, jams, puddings) or a high fat content
(dairy spreads, margarine, peanut butter), solid foods
(meat, fish, bread), and powders (flour, milk pow-
ders, spices). Frequently, finished foods are combi-
nations of these various groups. The approach used
in microscopic analysis depends upon the nature of
the food and the objective of the examination. The
history of food microscopy, and the principles of
microscopic analysis of different food groups were
reviewed comprehensively in a book by Aguilera and
Stanley in 1990 (see ‘Further Reading’ section). The
microscopic analysis of different food commodities
has been reviewed in books published in 1979 edited
by Vaughan and in 1994 edited by Flint (‘Further
reading’ section). From 1982 to 1994, the journal
Food Structure (formerly Food Microstructure) spe-
cialized in the publication of papers on food micro-
scopy and related techniques. Since 1994 there has
been no journal dedicated specifically to the micro-
scopic analysis of food, although the review journal
Trends in Food Science and Technology carries
regular articles on this subject.
Warley A (1997) X-ray Microanalysis for Biologists,
vol. 16, Practical Methods in Electron Micro-
scopy (series editor Audrey Glauert). London: Portland
Press.
Microscopy Techniques
Light Microscopy
Light microscopy (LM) is regularly used to obtain
rapid, inexpensive qualitative and quantitative infor-
mation in food analysis. The first routine use of LM
in food analysis was for the identification of adul-
teration (e.g., the presence of chicory root in coffee)
or contamination (insect, rodent, microbial, and
foreign bodies). Bright-field, polarizing, and fluores-
cent microscopy are the three traditional LM
techniques used most frequently in food analysis.
The basic instrument is a conventional compound
(bright-field) microscope, to which polarizing and
fluorescence accessories are easily attached.
A wide variety of stains can be used in combina-
tion with bright-field LM. In food analysis, however,
only a few of those stains are used on a regular basis.
For example, Toluidine Blue O (TBO) is a meta-
chromatic dye that produces different colors depen-
ding on the nature of the component to which it is
bound. Pectin-containing plant cell walls stain a rich
pink or purple with TBO, while lignified cells of
vascular tissues stain dark blue (see Figure 1A). TBO
is also useful for the examination of meat products,
where muscle fibers show up as pale pink, fibroblasts
as blue, and elastin fibers as turquoise. The familiar
blue coloration produced by staining starch with io-
dine is widely used, allowing the identification and
localization of starch even after processing has de-
stroyed the characteristic granule structure. For fat
staining, a lipid-soluble dye such as Oil Red O may
be used.
Polarizing microscopy is used to examine food
components that exhibit birefringence (an ordered
crystalline structure). Many food components are
birefringent, e.g., starch, plant cell walls, specialized
‘stone cells’ in some plant tissues, muscle fibers, fats
from both plant and animal sources, and different
types of flavor and seasoning components.
The botanical origin of most starches is readily
identified on the basis of granule size and shape,
the form and position of the hilum, and the
 MICROSCOPY APPLICATIONS / Food 51

(A) (B)

(C) (D)

Figure 1 Light microscopy of some foods of plant origin. (A) Bright-field micrographs of hand-cut section of fresh pumpkin stained
with Toluidine Blue. Parenchyma cell walls, which are rich in pectin, are purple, and cell walls of vascular tissue, which are rich in
phenolics, are blue. (B) Hand-cut section of fresh potato, viewed using polarizing optics. Birefringent starch granules appear bright
against a dark background. (C) Fluorescence micrograph of hand-cut section of pumpkin epidermis, stained with Coriphosphine O.
Bars in (A) to (C) represent 25 mm. (D) Fluorescence micrograph showing hand-cut section of infected radish, stained with Acridine
Orange (yellow fungal cells) and Methyl Green (green radish cell walls). Bar represents 100 mm. (Miller SS, Agriculture and Agri-food
Canada, Ontario, Canada.)
52 MICROSCOPY APPLICATIONS / Food
brilliance of the interference cross under polarized
light (see Figure 1B). Loss of birefringence upon
gelatinization, the temperature of which is charac-
teristic for various native and derivatized starches, is
determined using a polarizing microscope equipped
with a heat stage. The effects of baking and proc-
essing on starches are monitored by a combina-
tion of polarizing and bright-field microscopy
using iodine staining to identify starch after loss of
birefringence.
Fluorescence provides a sensitivity that is not
available in other forms of LM, allowing the detec-
tion of fluorescing compounds present in amounts as
little as 10 18 mol. A wide range of food compo-
nents of both plant and animal origin exhibit natural
fluorescence (autofluorescence). In plants, these com-
ponents include pigments such as chlorophylls and
carotenoids, as well as various phenolic compounds
such as lignins and low-molecular-mass compounds
such as ferulic and chlorogenic acids. Many flavor
compounds in herbs and spices are also autofluores-
cent. In animal tissues, the most common sources of
autofluorescence are bone, cartilage, collagen, elas-
tin, and some lipids. In addition, fluorescence may be
induced by a wide range of compounds such as flu-
orescent dyes, specific antibodies or lectins that are
conjugated to fluorescent markers, and substances
that fluoresce only in specific chemical environments.
For example, Calcofluor, a fluorescent brightener,
can be used as a highly specific probe to localize the
mixed linkage (1-3), (1-4)-b-D-glucan in cereal
grains. Nile red is a lipid-soluble dye that becomes
intensely fluorescent in the hydrophobic medium of
fat droplets. The precise identification and localization
of various components permitted by such compounds,
coupled with the sensitivity of the fluorescence tech-
nique, has made fluorescence microscopy a valuable
tool in food analysis.
Confocal microscopy has also been developed as a
technique with some advantages for food analysis.
The major difference between a confocal and a
conventional microscope is the placement of a pin-
hole at the focal plane of the image in the case of
the confocal instrument. This removes out-of-focus
light, generating a clearer image and allowing
optical sectioning of the specimen. There are two
basic types of confocal microscopy: confocal scan-
ning tandem microscopy (CSTM) and confocal
scanning laser microscopy (CSLM). CSTM uses
mercury, tungsten, or xenon illuminators and has
the advantage of allowing real-time observation
of the specimen. Low light intensity can be a prob-
lem with this technique. CSLM uses laser illumina-
tion. Standard equipment includes an argon laser
(488 and 514 nm wavelengths) with or without a
10 µm
Figure 2 Confocal laser scanning microscopy of Gouda
cheese stained for protein with 1-anilino-8-naphthalene sulfonic
acid. The depth resolution of optical sectioning was B0.7 mm.
(Courtesy of l. Heertje and Scanning Microscopy International.)
helium–neon laser (633 nm wavelength). In many of
the CSLM systems currently in use, real-time
observation is not available because of potential
damage to the eye by laser emissions. Images are
produced, stored, and manipulated by image-hand-
ling software. Light intensity is not a problem with
this method.
CSLM can provide focused images to a depth of up
to several hundred micrometers, depending on the
nature of the sample, so that sequential sections may
be obtained for three-dimensional reconstruction of
the image (Figure 2). In addition, several chemical
components (e.g., protein and fat in cheese) can be
identified and localized simultaneously using specific
fluorescent labels. CSLM has been used for the quan-
titative analysis of cellular structures in plant mate-
rial, the structural analysis of emulsions of different
complexities, and the location of microorganisms in
a wide range of food products.
Electron Microscopy
Electron microscopy (EM) is used where a higher
resolution than that obtainable by LM techniques is
required, for example, for the analysis of compo-
nents such as casein micelles in milk or pectin fibers
in fruits and vegetables, which are too small to be
visualized by LM. Scanning electron microscopy
(SEM) in conjunction with X-ray microanalysis,
and transmission electron microscopy (TEM) in the
electron energy loss spectrometry (EELS) mode, en-
able the analysis of foods or their components for
elemental composition. SEM of frozen hydrated
samples and TEM of replicas obtained from samples
fixed by rapid freezing provide images of foods un-
affected by chemical fixation. In contrast, chemical
fixation of certain components such as proteins
(using aldehydes – glutaraldehyde, formaldehyde –
and OsO4), unsaturated fats (using OsO4 in the
 MICROSCOPY APPLICATIONS / Food 53

Table 1 Overview of microscopical techniques and applications in food analysis

Technique Mode Information obtained

Dissecting microscopy Contamination

Light microscopy
Confocal microscopy
Scanning electron microscopy (SEM)
Energy-dispersive spectrometry
(X-ray microanalysis)
Transmission electron microscopy (TEM)
Electron energy loss spectrometry (EELS)
Atomic force microscopy
Bright-field, polarizing, and fluorescence
Confocal scanning tandem microscopy
(CSTM) and confocal scanning laser
microscopy (CSLM)
Conventional SEM
Cryo-SEM
Conventional SEM and cryo-SEM
Negative staining and metal shadowing
Thin sectioning
Freeze-fracture plus replication
Electron diffraction
Thin section TEM
Chemical and structural analysis
Contamination
Component identification
Component distribution
Changes during processing
3D reconstruction of low-density foods
Structural information for food powders,
solid foods, and viscous foods
Structural information for high-fat foods
and edible foams
Chemical information
Chemical and structural information for
liquid foods and emulsions
Chemical and structural information for
all foods
Chemical and structural information for
liquid foods, high-fat foods
Crystallinity
Chemical information
Very high resolution of surface
structure

presence of imidazole) and some polysaccharides
(using RuO4) followed by embedding in a resin and
staining of thin sections with heavy metals such as
uranyl acetate and/or lead citrate, makes it possible
to distinguish these components in foods.
has been widely used to study complex food
components, such as protein fibrils, polysaccharides,
and interfaces. Information concerning the different
types of microscopy and their applications is pre-
sented in Table 1.

Atomic Force Microscopy

Significant structural changes induced in food
ingredients by processing for EM require the
development of novel methods, such as environmen-
tal stages and metal-free processing, to achieve prop-
er results. Indeed, EM techniques are pushed to their
limits when they are used to image complex bio-
polymers such as the irregular fibrous proteins and
polysaccharides that are abundant in foods. Since
these types of samples have to be coated with metals,
the size of the metal grains restricts the level of detail
observable in the final images. Atomic force micros-
copy (AFM) creates an image by scanning a sharp
stylus, which is attached to a flexible cantilever,
across the sample surface. When the stylus is held
very close to the sample, repulsive forces deflect
the cantilever away from the surface. As the
cantilever–stylus assembly is scanned over the sur-
face, topological features are translated into movem-
ents, which can be recorded. This simple and
highly sensitive technique can measure deflections
caused even by individual molecules and atoms. It
Sample Preparation
Light Microscopy
Fresh material is examined wherever possible in
order to avoid artifacts (i.e., images resulting from
faulty preparation) and to prevent the loss of certain
components. Fresh material is also valuable where
time is an important constraint, as in quality control.
The examination of fresh material makes for the
most rapid and straightforward analysis.
Powders are examined in their original form by
dispersing them in either aqueous or nonaqueous
mountants, depending on their solubility and di-
spersibility. The mounting media may include stains
to increase the contrast of individual components for
structural observation or discrimination on the basis
of chemical and morphological characteristics. The
detection of birefringence in powders is also useful
for identification.
Smearing, squashing, and comminution are simple
preparation techniques that make it possible to
obtain extensive information quickly. Depending
54 MICROSCOPY APPLICATIONS / Food
on consistency, the product is mixed with appro-
priate dyes or is comminuted with a small amount
of water or dye before it is spread on a slide. The
sample may also be prepared for microscopy
by squashing between slide and coverslip. Using
these methods, appropriate dyes yield valuable
chemical information on the components of the
food, although the overall microstructure is gene-
rally lost.
Where information on chemical composition or
structural and spatial relationships is required (e.g.,
comminuted meat products), the sample is sectioned
before examination. The simplest and quickest pro-
cedure is to section the sample by hand using a
clean razor blade, but this is possible only with rigid
samples. Soft samples can be made rigid either by
freezing or embedding in paraffin or a plastic
resin such as glycol methacrylate. Frozen sections
are produced in a relatively short time and, with
the exception of ice crystals, have the advantage of
few artifacts. Ice-crystal damage can be minimized
by freezing the sample in nitrogen slush at 2051C
before mounting and sectioning in a cryostat. Rapid
work is essential, since ice crystals grow as the
sample warms up to the cryostat temperature
( 201C to 251C). Enzymatic activity in the sam-
ples is arrested at cryostat temperatures but can be
observed in situ using appropriate chromogenic or
fluorogenic substrates. With appropriate instrumen-
tation, it is even possible to measure the rate of
the reaction.
Fixation with formaldehyde or glutaraldehyde
before sectioning prevents the disintegration of
fragile sections on the slide when dyes or moun-
tants are applied. The use of slides coated with al-
bumin or poly-L-lysine to collect sections can be
helpful. Some components, particularly certain po-
lysaccharides, cannot be stabilized by fixatives, while
others of low molecular mass can migrate in the
sections or may be extracted from them when
dyes are applied. In such cases, vapor staining can
be used (e.g., starch with iodine vapor or fat
with OsO4 vapor). Embedding in a resin is carried
out when better resolution structural details are re-
quired. In such cases, sections 0.5–5 mm in thickness
are examined.
Electron Microscopy
Sample preparation for EM depends on the nature of
the food and the type of scanning or transmission
technique used. For SEM, dry foods are prepared in
the same way as powders. Solid foods based on pro-
teins (cheese, meat) are fixed in glutaraldehyde solu-
tions (1–3%, buffered near the pH value of the food),
dehydrated in ethanol, defatted by extraction with
chloroform or n-hexane, returned to ethanol, frozen
in liquid isopentane over liquid nitrogen or in nit-
rogen slush, and cryofractured. Since ethanol does
not form crystals on freezing, it is possible to freeze
ethanol-impregnated samples directly in liquid nit-
rogen. Dairy products treated in this way are
known to fragment spontaneously, but this does
not occur if the products are frozen in isopentane or
using Freons (although the latter are discouraged
due to their negative environmental impact). The
fragments are returned to ethanol and critical-
point dried. Dried fragments are mounted, coated
with pure or alloyed gold, and examined by SEM.
If fat is the food component of interest, samples
fixed in glutaraldehyde are postfixed with imidazole-
buffered OsO4 and the fat extraction step is
omitted. Viscous samples, which would disinte-
grate during sample preparation, are encapsulated
in agar gel. The capsules are treated in the same
way as solid samples. Cryofracturing provides
superior images in comparison with dry fracturing.
In hydrated food samples, however, crystals may
form during freezing resulting in artifacts. Only a
thin surface layer (20 mm) is free from such
crystals.
Cryo-SEM (examination at temperatures below
801C) is suitable for high-fat foods and foods of
plant origin such as cooked vegetables, which
could be altered by other methods involving dehy-
dration. Rapid freezing of the samples is essential for
the prevention of artifacts arising from ice-crystal
formation.
Low-voltage SEM is valuable when applied to fro-
zen uncoated samples because it allows the observat-
ion of food in the near-native (frozen hydrated) state
at a high magnification, particularly by field emission
SEM, which is best suited for this type of work. Since
the samples do not need to be coated with metal,
they may be fractured continuously over the course
of the observation.
In TEM, negative staining and metal shadowing
techniques are suitable for macromolecules (proteins,
polysaccharides) and their assemblies (micelles). The
TEM technique practiced most frequently is the
examination of thin sections. In order to examine
foods using this technique, small samples (o1 mm3)
are chemically fixed and embedded in a resin such
as Epon, Araldite, or Spurr’s. Dense foods such as
cheese, dough, comminuted meat products, seeds,
and extruded products require longer fixation and
impregnation times compared to porous foods.
Low-viscosity resins facilitate embedding of the
samples. Starch granules usually remain nonim-
pregnated. During sectioning, when the sections
 MICROSCOPY APPLICATIONS / Food 55

are floated on the surface of water, they swell and their crystalline forms, emulsions, and foams,
expand. Subsequent drying results in the develo- represent another important area of food structure
pment of folds in the granules. These result in studies in which all kinds of microscopy are
artifacts (dark, star-like areas in the micrographs). used. Findings from traditional foods may be ap-
Separate protocols have been developed for most plied to newly formulated foods to improve their
foods. consumer appeal.
Freeze-fracturing, freeze-etching, and replication
with platinum and carbon, followed by TEM of rep-
licates is the technique most often used for the ex- Dairy Products
amination of high-fat dairy products such as cream, Most dairy products, including yogurts and chee-
oil-in-water emulsions such as ice cream, and water- ses, are based on casein micelles (protein globules
in-oil emulsions such as margarines and low-fat B100 nm in diameter) and whey proteins. Casein
spreads. Micrographs obtained by any kind of mi- micelles and coagulated whey proteins must be ex-
croscopy, which used to be recorded on film, are now amined by EM because LM techniques do not
recorded as digitized images and may be manipulated provide sufficient resolution.
using computers and software, which come as part of Coagulation of milk results in two different kinds
the EM setup. of gel, depending on whether the milk has previously
been heated above 851C. Heating leads to interac-
tions between k-casein on casein micelle surfaces and
Applications
a whey protein called b-lactoglobulin. The resulting
Contamination of Food protein complex occupies sites suitable for interac-
tions with the micelles, and the gel consists of short
One of the most important applications of food mi-
interlinked chains and minute pores in which the
croscopy is the detection and identification of con-
liquid phase is firmly immobilized. This structure
taminants in food, usually following a consumer
retains the liquid phase and is important for soft milk
complaint. Macroscopic examination is followed
products such as yogurt. Large protein clusters
by examination under a binocular dissecting micro-
develop in coagula made from unheated milk, and
scope. Foreign bodies, such as metal, rubber/plastic,
result in syneresis of the gel, i.e., separation of the
glass, wood, and crystalline solids may be identified
liquid phase called whey. These structures are im-
at this stage, as also some biological contaminants
portant in cheese-making.
(e.g., animal or insect parts). Biological contaminants
Identification of specific food components such as
of microbial origin may not be detected at this stage,
proteins and polysaccharides is possible by immuno-
unless they have formed colonies. Some foreign bod-
localization using gold-labeled antibodies (Figure 3).
ies may occur in finished food because of crystalli-
In conjunction with X-ray microanalysis, SEM is
zation (e.g., tyrosine crystals in anchovy paste) or
used to analyze salt crystals in cheeses, including
interactions between components (e.g., calcium oxa-
late crystals in foods containing milk and vegetable
constituents). Sometimes food is unintentionally con-
taminated while being processed in the kitchen (e.g.,
by glass or enamel particles from kitchen utensils) or
even during consumption (e.g., by chips of tooth
enamel or fillings). Many of the more difficult cases
of food contamination can now be solved with the
aid of elemental X-ray microanalysis and using pro-
cedures compiled by Lewis (see ‘Further Reading’
section).

Structural Analysis
Food microstructure is intimately related to its sen-
0.5 µm
sory properties, e.g., elasticity, firmness, smoothness,
and juiciness. Proteins and carbohydrates form
Figure 3 Immunogold labeling of egg albumin (minute black
gels, the structures of which affect water- and
dots) which forms a wrapping layer around casein micelles (ar-
fat-retention properties. Gel structures of various rows) in microparticulated protein. The bar represents 0.5 mm.
biopolymers (e.g., gluten, myosin, starch, glycogen) (Reproduced from Singer and Dunn (1990) by N.S. Singer and
have been studied extensively. Fats and oils, John Wiley & Sons, Inc.; & 1990 Wiley.)
56 MICROSCOPY APPLICATIONS / Food
M used to analyze changes that occur in muscle struc-
C tures after the animal has been slaughtered, during
(a) 100 µm (b)
Ca P heating, which denatures proteins, leads to shrinkage
M
C
(c) 10 µm
Figure 4 Results of SEM-EDX analysis showing distribution of
(a) calcium and (b) phosphorus at Camembert cheese surface (c)
in the mass of mold hyphae M; C image of a freeze-fractured
Camembert cheese surface showing the protein matrix C and the
hyphae M. (Micrographs (a) and (b) reproduced by courtesy of
B.E. Brooker and Scanning Microscopy International.)
deposits of the surface of mold-ripened cheeses such
as Camembert. Metabolic activity inside the dense
hyphal mat at the cheese surface increases the pH of
the cheese and leads to the precipitation of calcium
and phosphate ions present in the cheese (Figure 4).
This process may continue until most of the ions
diffuse out of the cheese interior and form a deposit
at the cheese surface.
Cryo-SEM or TEM examination of platinum-and-
carbon replicas of whipped cream shows stabilizat-
ion of the air cells by fat globules that adhere to the
air–liquid interface. Distribution of intact fat glob-
ules and water droplets in butter and other emulsions
is also conveniently studied by these methods in
which the sample is fixed by rapid freezing.
Meats
The basic component of meat is muscle tissue, which
has a complex microstructure. Individual myofibers
are surrounded by connective tissue membranes
(endomysium) and bundles of myofibers are separa-
ted from each other by another connective tissue
membrane, the perimysium. Muscle also contains
veins, arteries, and nerve fibers. Skeletal (or striated)
muscle differs in microstructure from the smooth
muscle that is present in internal organs such as
intestinal walls, blood vessels, etc. Microscopy is
so-called rigor mortis and then during ageing, free-
zing, salting, and processing of the meat. These
processes cause changes to the muscle microstruc-
ture, but the most significant changes are caused by
of the muscle fibers, and the loss of fine structure.
Such changes can be observed by LM and EM tech-
niques. Individual components in raw meat, meat
heated to over 801C, and cooked meat can be de-
tected using specific antibodies to native and dena-
tured meat antigens in combination with quantitative
fluorescent microscopy.
Comminuted meat products consist of several
components and are usually referred to as meat
emulsions. The binding of meat and fat particles is
improved by the use of fillers, which consist of soy or
grain flours, sodium caseinate, starch, gluten, and
other polysaccharides and proteins. Their role is to
increase yield, improve stability, and modify textual
properties. Microscopy is used to detect the presence
of fillers, determine the quantity, and establish their
distribution.
Foods of Plant Origin
This group of foods is diverse, encompassing grain
flours, dough, bakery products, products derived
from starch and/or vegetable proteins, fruits, and
vegetables. The consistency of these foods ranges
from liquid (fruit and vegetable juices, beer, wine)
through moist foods (bread, baked potatoes, tofu) to
low-moisture products (potato crisps, bread-
crumbs). All these products are derived from plant
cells, the walls of which consist of cellulose, hemi-
celluloses, lignin, pectins, and proteins. The structure
and composition of plant cells as foods is discussed in
detail by Aguilera and Stanley (see ‘Further Reading’
section). Foods of plant origin are often prepared by
cooking and/or freezing and are amenable to analysis
by LM and EM techniques. In contrast, extruded
foods are produced by a complex process in which
high temperature (100–2001C), high pressure (2–
6 MPa), and mechanical shear are applied to the
food. The resulting porous as well as compact sam-
ples are difficult to embed in resin and to section for
LM and TEM. Therefore, SEM is the technique most
frequently used to analyze this category of food.
Compared to meat and dairy products, many plant
foods contain little or no protein. Starch cannot be
fixed by chemical methods, therefore, EM methods
involving freeze-fractured samples yield the best results.

Trends in the Microscopic Analysis of Foods
Over the last few years, there have been significant
advances in the analysis of food structures at the
microscopic level. These advances have been brought
about in part by the development of new techniques
and their application to traditional problems, but the
inclusion of image analysis as an essential component
of quantitative food analysis has also played an im-
portant role. Two of the most important develop-
ments are the increasing use of AFM and confocal
laser scanning microscopy in the analysis of food
microstructure. For example, AFM has been used to
investigate the structure of fats and oils, and has
provided convincing evidence for the presence of
microcrystal networks within such substances. This
was possible thanks to a recent innovation in AFM
technology, known as the tapping mode, in which the
cantilever connected to the stylus is oscillated as it
scans the sample surface, allowing AFM to be used
with soft and hydrated food materials. Another ex-
ample is the use of CLSM to study the impact of oil
frying on potato slices. The ability to generate non-
destructive optical sections of the sample facilitated
the detection of oil penetration into the crust, fol-
lowed by three-dimensional reconstructions that
showed the oil forming around the cells and pen-
etrating the food through intercellular spaces, while
almost no oil was seen in the cells themselves. It is
likely that these two key techniques will complement
LM and EM methods and play an increasingly
important role in food structural analysis in the
future.
Forensic
C S Palenik and S J Palenik, Microtrace, Elgin, IL, USA
& 2005, Elsevier Ltd. All Rights Reserved.
Introduction
The field of forensic microscopy is based upon
Edmond Locard’s Exchange Principle, which states
that when two objects come into contact, a transfer
of material will result. Although the amount of ma-
terial that is transferred in a contact may be minute,
microscopical examination can often reveal a great
amount of information regarding the materials that
came into contact and the way in which they made
contact. This article illustrates the way in which
forensic microscopy can capitalize on this transfer of
MICROSCOPY APPLICATIONS / Forensic 57
See also: Microscopy: Overview. Microscopy Tech-
niques: Light Microscopy; Electron Microscopy;
Specimen Preparation for Electron Microscopy; Scanning
Electron Microscopy; Atomic Force and Scanning
Tunneling Microscopy; X-Ray Microscopy.
Further Reading
Aguilera JM and Stanley DW (1990) Microstructural Prin-
ciples of Food Processing and Engineering. London/
New York: Elsevier.
Aguilera JM, Stanley DW, and Baker KW (2000) New di-
mensions in microstructure of food products. Trends in
Food Science and Technology 11: 3–9.
Ferrando M and Spiess WEL (2000) Confocal scanning
laser microscopy: a powerful tool in food science. Food
Science and Technology International 6: 267–284.
Flint O (ed.) (1994) Food Microscopy, Royal Microscop-
ical Society Microscopy Handbooks. Oxford, UK: BIOS
Scientific Publishers.
Kaláb M, Allan-Wojtas P, and Miller SS (1995) Microscopy
and other imaging techniques in food structure analysis.
Trends in Food Science and Technology 6: 177–186.
Kirby AR, Gunning AP, and Morris VJ (1995) Atomic-
force microscopy in food research – a new technique
comes of age. Trends in Food Science and Technology 6:
359–365.
Lewis DF (1993) A tutorial and comprehensive bibliograp-
hy on the identification of foreign bodies found in food.
Food Structure 12: 365–378.
Vaughan JG (1979) Food Microscopy. London: Academic
Press.
Wilkinson C, Dijksterhuis GB, and Minekus M (2000)
From food structure to texture. Trends in Food Science
and Technology 11: 442–450.
material to aid in establishing the facts within a
criminal investigation, describe the types of problems
that are addressed, the types of materials encoun-
tered, and the analytical approach used to extract
information from trace evidence. Finally, a practical
example that illustrates the extent to which micro-
scopic evidence can influence the outcome of a case
will be described.
The Role of Forensic Microscopy
The field of forensic microscopy is quite broad in that
it deals with any possible type of microscopic resi-
due, which may have biological, geological, or ant-
hropogenic origins. For example, in sorting through
the material encountered on a single pair of pants