Journal of Dentistry, 6, No. 4, 1978, pp. 327-341.

Printedin Great Britain

Replication techniques for the

scanning electron microscope
1. History, materials and techniques
I. E. Barnes, BDS, FDS”
Hard Tissue Unit, Anatomy Department, University College, London

ABSTRACT
This paper, which is presented in two parts, examines
microscope which are of value in dental research.
replication are reviewed, the assessment of techniques
replication techniques are discussed. The second part
and the interpretation of replicas.
replication techniques for the scanning electron
In the first part the history and principles of
is detailed and the materials suitable for use in
will deal with clinical and laboratory procedures

INTRODUCTION
The use of replicas in dental research is of long-standing. In 1940 Wolf studied teeth in vivo
by withdrawing a pellicle of celloidin from the enamel surface by means of an adhesive plastic
tape. The surface of the impression was illuminated by an obliquely sited light source and
viewed with an optical microscope. Eifmger (1964) described the use of cellulose
acetobutyrate applied in sheet form to a tooth surface that had been previously moistened
with acetone solvent.
With the development of the transmission electron microscope (TEM), replication
techniques developed by other disciplines were found to be useful for dental applications.
Richards and Thomassen (1944) used the method described by Heidenreich and Peck (1943)
whereby polystyrene was applied to the substrate surface at 2000-3000 lbf/in’ and 160 “C.
Silica was evaporated on to the polystyrene surface which was subsequently dissolved away
with ethyl bromide. Later workers utilized electron-lucent organic films shadowed obliquely
with metal (collodion/silver, Scott et al., 1949; collodion/aluminium, Charbeneau and
Peyton, 1957).
In the normal mode of use the scanning electron microscope (SEM) produces an image by
collecting secondary electrons emitted from the surface of the substrate, and there is no
reason to construct electron-lucent replicas. Replication techniques remain of value in dental
research, using the SEM, for short or long term in vivo studies (Facq and Volpe, 1970; Tay
et al., 1974), and also in order to prevent distortion occurring when teeth are viewed
directly.
Boyde (1969) observed and measured the expansion that occurred when dried elephant
dentine was rehydrated, and it is not surprising that such an elastic, organic and normally
wet material should thus distort. It is unlikely that enamel contractsmeasurably in vacuum, and
the complex inter-relationships that exist between enamel and dentine at the amelodentinal
junction (Orams et al., 1976) will tend to resist shrinkage of the underlying dentine. The

*Present address: Royal Dental Hospital of London School of Dental Surgery, London.
328 Journal of Dentistry, Vol. ~/NO. 4

Fig. 1. a, Unfilled cavity in the occlusal surface of a premolar from which both cusps have been cut. The
dentinal cracks continue into the enamel but, although closely related, they are discontinuous. Field
width 5 mm. b, Small crack at the margin of a cavity cut in dentine. This was not visible at low
magnifications. Field width 83 pm. c, Cutting edge of a microtome knife, prepared by fracturing plate
glass. (X2).

present author (unpublished) has been unable to measure any contraction in enamel-
supported dentine. It is, nevertheless, reasonable to suppose that enamel-supported dentine,
when subjected to vacuum, will develop internal stresses. When plane dentine surfaces
surrounded by enamel are viewed in the SEM, it is possible to observe the sudden relief of
internal stress by the development of cracks (Fig. la). Cracks appear most frequently when
the integrity of the dentine has been weakened by cavity preparation, and they may be
accompanied by a contraction of the dentine from the amelodentinal junction. Smaller
cracks may be seen at the cavity margins but may not be noticed unless sought (Fig Ib).
Cracks also develop in the enamel and may or may not be continuous with those formed
in the dentine (Fig. Ia). The enamel cracks are probably naturally occurring defects that
have been widened owing to the distortion of the underlying dentine or by cavity prepara-
tion. Natural enamel cracks are commonly found and are present before the tooth erupts
Barnes: Replication techniques for SEM 329

(Scott et al., 1949). Cracks developing during cavity preparation have been investigated by
Despain et al. (1974) who used a replication technique. Haines et al. (1963) demonstrated
the compressibility of enamel and related this to the 2 per cent water content. Jorgensen et al,
(1976) measured deformation of Class III and IV cavities in a tooth subjected to occlusal
loading, and it is possible that deformation of enamel also occurs (through the lines of
weakness formed by developmental cracks) when the underlying dentine becomes stressed
as a result of water loss in vacuum. Such deformation may cause interfacial deficiencies
between the cavity wall and restoration to be misinterpreted as polymerization shrinkage,
and most workers acknowledge that replication techniques should be used for studies on
marginal adaptation (Bergvall and BrZlnnstrbm, 1971; Grundy, 1971; Sela et al., 1975;
Barnes, 1977).

PRINCIPLES AND NOMENCLATURE

The principles of replication are well understood but at present there seems to be no
universally accepted system of nomenclature. This confusion stems from the replication
techniques used in transmission electron microscopy, where the impression is looked through
and thus yields a positive image of the substrate. In such a situation it is justifiable to call the
impression a replica. When an impression is viewed in the SEM it is looked at, not through,
and since it represents a negative image of the substrate it cannot be called a replica. The
term ‘primary impression’ is sufficient, whether it is to be examined directly in the SEM or
used as the first stage in the construction of a cast replica. It is desirable to reserve the term
‘negative replica’ for a further stage in model construction, which will be described later. A
fluid casting material is usually introduced into the primary impression and when set is
viewed in the SEM. This cast is, hopefully, an accurate facsimile of the substrate, and is
correctly termed the ‘replica’.

Negative replica
In some fields of research, for example palaeontology, the construction of negative
replicas is of value, and it is felt that negative replicas may have a place in dental research
(see below).
The imprints of ancient microscopic life forms may remain in rocks in the form of empty
undercut channels. Casts of such channels will reveal the structure of the original organism,
but, unless elastic, can be released only by solution of the surrounding rock. A solid cast
can, however, be constructed without destroying the substrate if a three-stage technique is
employed. The stages are: primary impression, secondary impression of the primary
impression and a cast made from the secondary impression. The final cast will be a ‘negative
replica’ of the original rock substrate and a solid facsimile of the original organism (Fig. 2).

ASSESSblENT OF REPLICATION TECHNIQUES

When impressions are to be studied in the SEM the prime consideration is the ability to
reproduce fine surface detail. If, however, replicas are to be produced, it is necessary first to
establish the compatibility of the impression and casting materials. A simple screening method
has been found to be effective.
Screening technique
The ability of two materials to replicate a glass microtome knife (Fig Zc) has been found to be a
quick and sensitive method of screening the accuracy and compatibility of combinations of
330 Journal of Dentistry, Vol. ~/NO. 4

Substrate (a)

Primary
Impression(b)

Replicaor
secondary
impression k)

Negative
replica (0)

Fig. 2. Primary impression (bl can be viewed in

the SEM or used as a mould from which a replica
(cl is cast. The replica is a rigid facsimile of the
substrate (a). A secondaw impression can also be
made from the primary impression, and when
resin is cast into this a rigid negative replica (d)
is produced.

impression and casting materials. (Knife edges were originally used in this way to obtain
silicone rubber moulds for standardized wax patterns in an investigation on cast gold knife
edges.) The plane (unfractured) sides of the cast knife edge may be examined with the
naked eve for shrinkage, warp or loss of sheen, and with a sharp probe for a tacky unset
surface. The ability to reproduce a microscopically smooth surface can be assessed by
examining the replicated fractured surfaces in the SEM; similarly, the replicated knife edge
may be compared with the glass original and is a good test of the ability to reproduce fine
detail. The adhesion or lack of adhesion of the casting material to the impression can be
readily determined in the knowledge that mechanical interlocking into gross surface
irregularities can be discounted. The ability to reproduce a smooth surface seems to be a
subtle test of a replication system, and even the Silflo/epoxy resin system that has proved
satisfactory for replicating teeth and restorations has, to date, yielded imperfect replicas of
fractured, smooth glass surfaces. This may be due to the inability of the resin to wet the
mould.

Assessment of surface detail

The ability of an impression material to resolve surface detail may depend not only on the
properties of the material but on the direction and magnitude of the force that is applied
to aid its conformation. The nature of the substrate may also be of importance, smooth
surfaces, such as glass, being unconducive to flow, and rougher surfaces aiding adaptation by
interlocking or capillary flow.
A premolar crown in which a Class I amalgam has been placed has proved to be a good
substrate for testing replication. The crown is cleaned of organic debris by immersion in
Barnes: Replication techniques for SEM 331

sodium hypochlorite solution. The enamel surface provides numerous areas of fine anatomical
feature, which lie at different angles to the path of application of the impression material
(normal at the cusp tip, tangential at the sides of the cusp). The ability to flow into and
reproduce irregular surfaces can be assessed on the amalgam filling. If a standard casting
resin is used, the accuracy of different impression materials can be assessed, or if an optimal
impression is used, casting materials can be similarly tested. It must be noted that in any
series of tests it is necessary to apply the impression materials in varying sequence, so that
loss of surface detail due to contamination of the substrate by previous impression materials
may be allowed for and discounted.

Mechanical properties of impression materials

If an impression material has a low tensile strength, or is inelastic, it may tear when it is
withdrawn from undercut areas. Tearing is an important source of artifacts when the
marginal adaptation of fillings is being studied by replication techniques. This will be
discussed in more detail in the second part of this work.
The ability of an impression material to withdraw from etched enamel or the exposed
resin tags of a composite restoration applied to etched enamel is a good indication of its
ability to resist tearing. A trial substrate can be prepared by placing a composite restoration
in an etched Class V cavity and subsequently eroding the surrounding enamel wall with acid
for 10 minutes, so as to expose the resin tags. It is difficult to compare different materials
when this technique is used, since torn impression material is frequently retained on the
substrate and will prevent that area from being assessed in subsequent replicas. Normally,
however, a number of areas will remain unobscured throughout a series of impressions.

IMPRESSION MATERIALS
Several workers have continued to use the organic films used in transmission microscopy for
their SEM investigations (vinyl resin/acetone primary impression, Galil and Gwinnett, 1975;
Galil, 1976; acetate tape/acetone primary impression, Silflo replica, Pameijer and Stallard,
1972a). Such films may yield good surface detail but cannot reproduce the larger contours
of the specimen. A method has been described (Barnes, 1972) whereby polyvinyl formal
may be withdrawn from the substrate surface in three-dimensional form; nevertheless, the
majority of workers now use some form of filled synthetic rubber as the primary impression
material. Sampson (1961) described the use of a Silflo primary impression cast in
nitrocellulose to make replicas for optical microscopy. Techniques using Silflo as the primary
impression have also been described by Sarkany (1962), Facq and Volpe (1970), Johnson
et al. (1970), Grundy (1971), Al-Hamadani and Crabb (1975) and Barnes (1977). Despain
et al. (1974) Tronstad and Leidal (1976) and Luescher et al. (1977) used Xantopren blue.
Condon and Harcourt (1977) described the use of the polyether rubber Impregum, both
with and without the use of thinner. A comprehensive article by Kusy and Leinfelder
(1977) recommended Xantopren blue and Silene of the 14 materials that were tested.
A series of impression materials has been investigated by the author and will now be
discussed.

Polysulphide rubber
Gmndy (1971) noted that polysulphide rubbers do not yield replicas of sufficiently fine
detail. In the author’s experience it is impossible to separate set epoxy resin replicas from
polysulphide (Permalastic) rubber base impressions, even if the impression is coated with
fig. 3. Effect of impression viscosity upon the replication of surface detail. a, Cut enamel surface used as
a control substrate for the assessment of replica impression materials. Field width 83 pm. b, Replica
constructed from epoxy resin cast into Silflo impression material. The substrate is well replicated. Field
width 91 pm. c, Replica constructed from epoxy resin cast into Lastic 55 impression. The detail is poten-
tially good but the surface is marred by a granular roughness. This may be due to the readiness with which
Lastic 55 tears when applied to undercut or microscopically rough surfaces. Field width 91 Mm. d, Replica
constructed in epoxy resin from a Cardex-Super impression. Surface detail is poorly reproduced. Field
width 91 IUD. e, Replica of an unpolished amalgam filling cast in epoxy resin from a Silflo impression.
Field width 50 Mm (approx.). f, Replica of the same part of the unpolished amalgam seen in e. The
impression was made in viscous Lastic 55, but the detail is similar to that obtained with the more fluid
Silflo. Field width 50 pm (approx.).
Barnes: Replication techniques for SEM 333

Fig. 4. Viscous impression material /a/ is applied

with vertical pressure 161 to an irregular substrate
(cl. The applied force will react at the horizontal
interface and will result in good adaptation of the
impression material. Less force will be transmitted
to the vertical interface and the adaptation will be
poorer.

gold before the resin is cast. This would suggest that the retention may be due in part to
microporosities in the rubber.
Silicone rubber
Four silicone rubber impression materials of different viscosities have been assessed and
compared in a series of experiments: Cardex-Super, putty-like; Lastic 55, viscous paste;
Silflo, fluid paste; Verone Perfecting Paste, very fluid paste. The Araldite replicas cast in the
Verone impressions set with a tacky surface, and it was considered that the Perfecting Paste
may have contained a plasticizing agent that leached into the setting resin.
Surface de tail
Usually the surface detail of the replicas increased as the viscosity of the impression
material decreased (Fig. 3~4). However, this was not an invariable finding and a number
of the Cardex-formed replicas showed areas of fine detail, indistinguishable from Silflo-
formed replicas. Regions of comparable accuracy were seen even more frequently in the
Lastic 55 and Silflo-formed replicas (Fig. 3e, fl.
It is probable that viscous impression materials are capable of good adaptation if strong
adaptational pressure is directly applied. In the case of the replica shown in Fig. 3f, the
Lastic 55 was held within a copper ring and considerable pressure was applied to the impres-
sion material at the upper open end of the ring during the initial application to the tooth.
When a very viscous material, such as Cardex, is applied from above, most of the force of
application will react against the horizontal interface, but less force will be transmitted to
the vertical interface. Consequently the occlusal adaptation will be good, but the detail
on the vertical walls will be poorly reproduced (Fig 4).
The impression material of choice should, therefore, be relatively fluid, since it is not
always possible or convenient to apply pressure, and if pressure is applied, it may not re-
act equally at all interfaces.
 Fig. 5. a, Replica of an etched enamel surface (left) and a Class V composite filling with exposed resin
tags (top right). The Lastic 55 impression material has adhered to the etched enamel and has torn when
the impression was withdrawn from the tooth or the replica. Large Lastic 55 filler particles are visible on
the enamel surface. Field width 166 pm. b, Surface of set Lastic 55 impression material which has been
torn experimentally. Filler particles of widely varying sizes constitute a large part of the material and
are probably the cause of its low tensile strength. Field width 100 hrn. c, Torn surface of a set Silflo
impression. The filler particles are very small and the torn surface appears finely granular. Field width
100 @m. d, Torn Cardex-Super impression material. The torn surface appears smooth when compared
with the fine granularity of torn Silflo. The small interface irregularities may be filler particles that re-
main covered with a thin layer of rubber. Field width 83 pm. e, Cut dentine surface that was used as a test
substrate. Field width 100 pm,. f, Replica of the substrate shown in e. The replica was cast in epoxy resin
from a polyether (Impregum) impression. The surface detail of the replica corresponds closely to that of
the substrate. Field width 83im1.
Barnes: Replication techniques for SEM 335

Mechanical properties
It was found during the experiments discussed above that Lastic 55 frequently tore when it
was withdrawn from etched enamel surfaces or tagged composite margins (Fig. AZ). The torn
silicone was retained either on the substrate or on the replica, but in both cases the
replicated surface was partly obscured. The surface of the torn impression revealed large
numbers of coarse filler particles (Fig. AZ, b) and it is probable that these caused the
friability of the material. Silflo tore less frequently, and examination showed the filer
particles to be extremely fine (Fig. 5~). Cardex-Super tore least of all, partly because its
imperfect adaptation reduced the keying effect, but also, it is felt, because of its homogeneity
(filler particles could not be positively identified, Fig. 5d).

Polyether rubber
Impregum was assessed in another series of experiments and gave good results (Fig. 5e, j),
although a number of the replicated surfaces appear to be rather granular. The material is
sticky and is more difficult to handle than the light-bodied silicones. In addition, the
stickiness predisposes to the formation of replicated air-bubbles. These problems may be
overcome by the use of thinners as described by Condon and Harcourt (1977). A further
disadvantage is that polyether rubber is incompatible with epimine resin casting materials
(see below).

Hydrocolloid
Pameijer and Stallard (1972b) have used hydrocolloid as a mucostatic impression material for
the replication of mucous membranes. The replicas were cast in silicone rubber.

Direct examination of the primary impression

Galil and Gwinnett (1975) examined vinyl resin primary impressions in the SEM in order
to assess the shape of fissures in molar teeth. Several authors have studied the resin tags at
the surface of freed composite restorations and have thus identified the etch patterns of
the enamel substrate. In this situation the composite filling may be considered to be a
primary impression of the etched enamel.
In general, direct examination of the primary impression is problematical for two
reasons. First, it has been the author’s experience that when gold is coated directly into
silicone impressions it may craze (Fig 6b, c). Grundy (1971) suggested that this may be
caused by the distortion of the silicone rubber owing to its high coefficient of thermal
expansion. Thermal changes may occur whilst coating or viewing the specimen. Other
workers have coated and viewed rubber-based materials either as primary impressions or as
cast replicas and have not noted crazing (Pameijer and Stallard, 1972b; Tay et al., 1974;
Brennan et al., 1975). Kusy and Leinfelder (1977) emphasized the importance of reducing
heat production during coating, and when viewing in the SEM.
Secondly, interpretation of the primary impression is difficult. The use of the negative
image facility provided on some microscopes will not fortuitously transform the image of
the impression to that of its substrate (Figs. 6, 7). For most purposes it is necessary to
construct a replica.

THE REPLICA
Although silicone rubber has been used as a casting material, replicas are usually constructed
from resins, the epoxy resins being the most commonly used (Araldite, Johnson et al.,
 336 Journal of Dentistry, Vol. ~/NO. 4

Fig. 6. Interpretation of negative impressions. a, Replica of a Class III silicate filling in an extracted
incisor tooth (Silflex/epoxy resin). Field width 2 mm. b, Impression from which the replica seen in a
was prepared. The impression has subsequently been gold-coated. The gold coating has crazed, either
during the coating procedure or whilst the specimen was in the SEM. Air-bubbles seen replicated on the
surface of the silicate filling are represented here as black voids. Field width 4 mm. c. Gold-coated
impression seen in 6, showing the lower part of the field at greater magnification. The SEM has been
used in the negative image mode. Although the crazing makes interpretation difficult, it can be seen that
the negative mode does not transform the voids formed by the air-bubbles into images of air-bubbles.
Field width 1 mm.

1970; Grundy, 1971; Barnes, 1972; Epoxylite EV4, Condon and Harcourt, 1977; Spurr’s
resin, Al-Hamadani and Crabb, 1975).
McLean and Wilson (1974) illustrated the use of an epimine resin, Scutan, although it is
uncertain whether this material was used for the primary impression or as the casting
medium. Further work confirming the dimensional and morphological accuracy of Scutan
as an impression and casting material has been undertaken by Cowell and Saxton (1978).
The resins discussed above are applied in the unpolymerized state and allowed to set
within the primary impression. Facq and Volpe (1970) described the use of resin pellets
(_Versalon 1112, polyamide resin) that were melted at 275 OF before casting into the
impression.
Barnes: Replication techniques for SEM 337

oblique lighting
from behind

Fig. 7. Interpretation of impressions and impressions

viewed with a negative image. a, Substrate illumin-
ated from above and behind. b, Impression of
substrate illuminated from above and behind. c,
Impression of substrate illuminated from above and
behind, but viewed with the negative image facility.
There is no similarity between the image thus
produced and that of the original substrate.

Assessment of casting materials

The assessment tests described earlier have been used by the author to investigate a number
of potential casting materials, of which three will be discussed. The materials were:
Durcupan ACM, epoxy resin embedding agent; Britfm, epoxy resin cement; Scutan,
epimine resin temporary crown material.
Scufan epimine resin is a paste/liquid catalyst system that is used in clinical dentistry for
the construction of temporary crowns. For replica construction one drop of catalyst was
mixed with 4 cm of the paste in order to retard the set. The viscous mixture was applied to
the silicone impression with a spatula, and was conformed to the surface with ajet of air from a
hand-held chip syringe. Scutan adheres to polyether rubber and cannot therefore be used
with Impregum impressions.
Britfa epoxy cement is an inexpensive household adhesive and was chosen because of its
relatively low viscosity. It is a paste/paste system and was applied to the impression in the
same way as the Scutan.
Durcupun epoxy embedding resin will be discussed in detail in the second part of this
paper.
The cast materials were allowed to set at SO’C for the following periods of time:
Durcupan 48 hours, Britfix 5 hours and Scutan 10 minutes. These times are nominal and
may well be capable of reduction.
At magnifications of x 1000 there was little difference between the three materials.
Above this magnification the Britfur replicas were inferior, but no differences were seen
between the Durcupan and Scutan replicas up to the maximum assessment value of x 5000
(Fig. 8a-6). In some situations the opacity and viscosity of Scutan were a disadvantage. For
example, when attempts were made to replicate the pin that is shown in Fig. 8e,it was not
possible to puddle Scutan down the pin hole in the impression, although this was readily
Fig. 8. Effect of different casting materials upon the replication of surface detail. a, Replica of an
unpolished amalgam filling cast in epoxy resin from a Silflo impression. Field width 42 &ml. 6, Replica
cast in epimine resin (Scutan) into the impression that was used to produce the epoxy replica seen in a.
The detail appears to be identical in both cases and is limited by the accuracy of the impression itself.
Field width 42 Mm.=, Replica of etched tooth surface. The ribbon-like structure was composite resin that
had infiltrated a crack in the enamel and which was revealed when the anamel was etched. Field width
83 pm. d, Replica cast in epimine resin (Scutan) from the impression used to produce the replica seen in
c. The reproduction of detail is again limited only by the accuracy of the impression. Field width
83 pm. e, Threaded pin that had been inserted into dentine. Field width 5 mm. ( Threaded pin seen in e
replicated in epoxy resin from a Silflo silicone impression. Field width 910 pm.
Barnes: Replication techniques for SEM

Fig. 9. a, Resin tags at the surface of an Adaptic composite restoration that has been adapted to etched
enamel. The restoration was isolated by dissolving the enamel in acid. The composite resin is a negative
replica of the enamel surface to which it was adapted. Field width 100gm. b, Replicaof an enamel surface
prepared by casting epoxy resin into a gold-coated silicone rubber impression. The gold coating has
separated from the impression and has withdrawn on the surface of the set replica. At some time during
the preparation of the replica the gold has become wrinkled. Field width 500 pm.

done with epoxy resin (Fig. Sf). Scutan is expensive, but the advantages of being able to
prepare a viewable replica from the primary impression in 10 minutes is apparent.

Other materials
Polyester resin: Isopon polyester resin was cast into the silicone knife edge impressions.
Polymerization shrinkage and unset surfaces precluded further investigation. Similar findings
have been reported by Barker and Parsons (1965). Composites: The composite resin tags
formed against etched enamel have been studied extensively (Fig. 9~). Because of the fine
surface detail exhibited by these tags, it seemed reasonable to suppose that composite resins
would make convenient casting materials. Filled composites, resins and glazes were applied
to silicone impressions, and in every instance a thin layer of unset resin remained on the
surface of the replica.

Gold-coating the replicas

It is possible to cast epoxy resin into a gold-coated primary impression and withdraw the
gold on the surface of the set replica. In theory this should lead to a higher degree of
dimensional accuracy. In the author’s hands this technique has proved to be impractical as
the coating crinkles and becomes tesselated (Fig. 9b). The thickness of the gold coating that
is evaporated on to the replica surface is about 30 nm, which is of little significance at the
magnifications at which the replicas are examined. Replicas may be gold-coated by
sputtering or evaporation in vacuum. When sputter-coating, the unit should be run in several
short bursts, and when vacuum-coating the replicas should be a sufficient distance from the
filament to prevent melting of the replica due to overheating.

REFERENCES
Al-Hamadani K. K. and Crabb H. S. M. (1975) Marginal adaptation of composite resins. J.
Oral Rehabil. 2, 21-33.
340 Journal of Dentistry, Vol. ~/NO. 4

Barker B. C. W. and Parsons K. (1965) Replicas of teeth. Aust. Dent. J. 10, 392-394.
Barnes I. E. (1972) Replica models for the scanning electron microscope. A new impression
technique. Br. Dent. J. 133,337-342.
Barnes I. E. ( 1977) The adaptation of composite resins to tooth structure. Part 1, Study 1:
Introduction and the adaptation of composite resins to the unetched enamel cavity wall.
Br. Dent. J. 142, 122-129.
Bergvall 0. and Brinnstrom M. (197 1) Measurements of the space between composite resin
fillings and the cavity walls. Sven. Tandlak. Tidskr. 64,217-226.
Boyde A. (1969) Correlation of ameloblast size with enamel prism pattern: use of scanning
electron microscope to make surface area measurements. Z. Zellforsch. Mikrosk. Anat.
93,583-593.
Brennan S., Condon G., Dickinson C. et al. (1965) Scanning electron microscope study of
margins of restorations. J. Dent. Res. 54,644.
Charbeneau G. T. and Peyton F. A. (1957) Observations from shadowed collodion replicas
of teeth with amalgam restorations. J. Dent. Res. 36,623-63 1.
Condon G. D. and Harcourt J. K. (1977) Modified in vivo replica technique for scanning
electron microscopy. J. Dent. Res. 56, 542.
Cowell C. R. and Saxton C. A. (1978) Assessment of non-elastomeric polymer for the
replication of tooth surface in vivo. J. Dent. 6, 210-216.
Despain R. R., Lloyd B. A. and Brown W. S. (1974) Scanning electron microscope investiga-
tion of cracks in teeth through replication. J. Am. Dent. ASSOC. 88, 580-584.
Eifinger F. F. (1964) Replica technic in the oral cavity. Dent. Abstr. 9,227-228,
Facq J. M. and Volpe A. R. (1970) In vivo actual abrasiveness of three dentifrices against
acrylic surfaces of veneer crowns. J. Am. Dent. Assoc. 80, 317-323.
Galil K. A. (1976) Accurate replication of enamel for examination by scanning electron
microscopy. J. Dent. Res. 55, 710.
Galil K. A. and Gwinnett A. J. (1975) Three-dimensional replicas of pits and fissures in
human teeth: scanning electron microscopy study. Arch. Oral. Biol. 20,493-495.
Grundy J. R. (1971) An intra-oral replica technique for use with the scanning electron
microscopy. Br. Dent. J. 130, 113-117.
Haines D. J., Berry D. C. and Poole D. F. G. (1963) Behaviour of tooth enamel under load.
J. Dent. Res. 42, 885-888.
Heidenreich R. D. and Peck V. G. (1943) Fine structures of metallic surfaces with the
electron microscope. J. Appl. Physics 14, 23-29.
Johnson C., Dawber R. and Shuster S. (1970) Surface appearances of the eccrine sweat duct
by scanning electron microscopy. Br. J. Dermatol. 83,655-660.
Jorgensen K. D., Matono R. and Shimokobe H. (1976) Deformation of cavities and resin
fillings in loaded teeth. Odontol. T. 84, 46-50.
Kusy R. P. and Leinfelder K. F. (1977) In situ replication techniques. I: Preliminary screen-
ing and the negative replication technique. J. Dent. Res. 56, 925-932.
Luescher B., Lutz F., Ochsenbein H. et al. (1977) Microleakage and marginal adaptation in
conventional and adhesive Class II restorations. J. Prosthet. Dent. 37, 300-309.
McLean J. W. and Wilson A. D. (1974) Fissure sealing and filling with an adhesive glass-
ionomer cement. Br. Dent. J. 136,269-276.
Orams H. J., Zybert J. J., Phakey P. P. et al. (1976) Ultrastructural study of human dental
enamel using selected-area argon-ion beam thinning: Arch. Oral. Biol. 21, 663-675.
Pametier C. H. and Stallard R. E. (1972a) Application of replica techniques for use with
scanning electron microscopes in dental research. J. Dent. Res. 51, 672.
Pameiier C. H. and Stallard R. E. (1972b) A pressureless replica technique for use with the
scanning electron microscope. J. Dent. Res. 5 1, 1680.
Barnes: Replication techniques for SEM 341

Richards A. G. and Thomassen L. (1944) Microstructure of tooth surfaces as revealed by the

electron microscope. J. Am. Dent. Assoc. 31, 772-776.
Sampson J. (1961) A method of replicating dry or moist surfaces for examination by light
microscopy. Nature (Lond.) 191,932-933.
Sarkany I. (1962) A method for studying the microtopography of the skin. Br. J. Dermatol.
74254-259.
Scott D. B., Kaplan H. and Wyckoff R. W. G. (1949) Replica studies of changes in tooth
surfaces with age. J. Dent. Res. 28, 31-47.
Sela M., Sela J., Arad T. et al. (1975) Adaptation of silicate and Adaptic to the margins of
cavities. A scanning electron microscope study. J. Oral Rehabil. 2, 117- 124.
Tay W. M., Waite I. M., Morrant G. A. et al. (1974) An assessment of anterior restorations
in vivo using the scanning electron microscope. Results after one year. Br. Dent J. 137,
463-47 1.
Tronstad L. and Leidal T. I. (1976) New instrument for finishing of embrasure margins of
Class II cavities. J. Am. Dent. Assoc. 93, 94-97.
Wolf J. (1940) Plastische histologie der zahngewebe. Dtsch. Zahn. Mund. Keiferheilkd. 7,
265-284.