Ultrasound in Med. & Biol., Vol. 44, No. 12, pp.

24922504, 2018
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https://doi.org/10.1016/j.ultrasmedbio.2018.07.010

Review Article

MUSCLE ARCHITECTURE ASSESSMENT: STRENGTHS, SHORTCOMINGS AND NEW

FRONTIERS OF IN VIVO IMAGING TECHNIQUES

TAGEDPD1X XMARTINO V. FRANCHI,D2X X*,y D3X XBRENT J. RAITERI,D4XzX D5X XSTEFANO LONGO,D6XxX D7X XSHANTANU SINHA,D8X{X
D9X XMARCO V. NARICI,D10X║X and D1X XROBERT CSAPOD12#X X TAGEDEN
* Laboratory for Muscle Plasticity, Balgrist University Hospital, University of Zurich, Zurich, Switzerland; y Sports Medicine
Research, Balgrist University Hospital, University of Zurich, Zurich, Switzerland; z Faculty of Sport Science, Ruhr University
Bochum, Germany; x Department of Biomedical Sciences for Health, Universit
a degli studi di Milano, Milan, Italy; { Department of
Radiology, University of California San Diego, San Diego, California, USA; ║ Institute of Physiology, Department of Biomedical
Sciences, University of Padua, Padua, Italy; and # Research Unit for Orthopaedic Sports Medicine and Injury Prevention, Institute for
Sports Medicine, Alpine Medicine & Health Tourism (ISAG), University for Health Sciences, Medical Informatics and Technology,
Hall, Austria
(Received 27 February 2018; revised 10 July 2018; in final from 13 July 2018)

Abstract—Skeletal muscle structural assembly (and its remodeling in response to loadingunloading states) can
be investigated macroscopically by assessing muscle architecture, described as fascicle geometric disposition
within the muscle. Over recent decades, various medical imaging techniques have been developed to facilitate the
in vivo assessment of muscle architecture. However, the main advantages and limitations of these methodologies
have been fragmentally discussed. In the present article, the main techniques used for the evaluation of muscle
architecture are presented: conventional B-mode ultrasonography, extended-field-of-view ultrasound, 3-D ultra-
sound and magnetic resonance imaging-based diffusion tensor imaging. By critically discussing potentials and
shortcomings of each methodology, we aim to provide readers with an overview of both established and new tech-
niques for the in vivo assessment of muscle architecture. This review may serve as decision guidance facilitating
selection of the appropriate technique to be applied in biomedical research or clinical routine. (E-mail: martino.
franchi@balgrist.ch) © 2018 World Federation for Ultrasound in Medicine & Biology. All rights reserved.
Key Words: Skeletal muscle, Morphology, Structure, Sonography, Pennation angle, Fascicle length, Muscle
thickness.

INTRODUCTION
Up to the early 1990s, muscle architecture, defined as the
physical arrangement of muscle fibers, was studied in
cadaver specimens (Huijing 1985; Wickiewicz et al.
1983). Such measurements were used to assemble com-
prehensive databases (Lieber and Friden 2000) that pro-
vide the backbone of our understanding of muscle
structure and its functional importance. Ex vivo studies
also feature the advantage of facilitating the study of
architectural parameters at the cellular scale (e.g., sarco-
mere length), which are below the resolution limits of all
non-invasive imaging-based in vivo techniques. Yet,
cadaver-based measures are dependent on the premortem
Address correspondence to: Martino V. Franchi, Laboratory for
Muscle Plasticity and Sports Medicine Research, Department of Ortho-
pedics, Balgrist University Hospital, University of Zurich, Zurich,
Switzerland. E-mail: martino.franchi@balgrist.ch
age and health status of patients and may be affected by
perimortal changes and fixation artifacts. Further, cadaver-
based measures are limited in that the dynamics of muscu-
lar contraction cannot be studied.
To overcome these limitations, medical imaging
techniques have been applied to study the in vivo muscle
architectural properties at the tissue scale. In particular,
brightness mode (B-mode) ultrasound imaging has been
used extensively for more than 25 y (Rutherford and
Jones 1992; Selva Raj et al. 2017) and represents the
technique most frequently applied to quantify skeletal
muscle mass and architectural features and relate these
parameters to muscle function (Lieber and Friden 2000;
Narici 1999). In the last 15 y, more refined methodolo-
gies, such as extended-field-of-view (EFOV) and 3-D
ultrasound as well as diffusion tensor magnetic reso-
nance imaging (DTI), have been developed and imple-
mented for the study of muscle architectural properties.

2492
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al.
These techniques feature distinct advantages but also
significant shortcomings. Our aim was to review the
principles underlying the current methodologies used for
the assessment of muscle architecture and critically dis-
cuss their strengths, limitations and potential future
directions. By providing readers with an overview of
both established and new techniques for the in vivo
assessment of muscle architecture, this review may serve
as decision guidance facilitating selection of the appro-
priate technique to be applied in biomedical research or
clinical routine.
B-MODE ULTRASOUND IMAGING
Principles
Conventional B-mode ultrasound can provide both
quantitative and qualitative information on skeletal mus-
cle (Ticinesi et al. 2017), tendons and ligaments. In this
section, we discuss only conventional muscle architec-
ture evaluation through B-mode ultrasound. For an
extensive review of further ultrasound-based techniques
used to determine various tissue mechanical properties,
we refer the reader to Lichtwark (2017).
B-Mode ultrasound relies on the transmission of
echo pulses from an ultrasound transducer. The capac-
ity of ultrasound waves to penetrate the body is directly
proportional to their wavelength, whereas image spatial
resolution is inversely proportional to it. For the study
of skeletal muscles, frequencies of 510 MHz (and, on
rare occasions, up to 17.5 Hz for superficial muscles)
are most commonly used. As ultrasound waves pene-
trate the body, they pass areas distinguished by differ-
ent acoustic impedance—lowest for air and highest for
bone (Chan et al. 2008)—and are partly reflected back
to the transducer. The reflection coefficient (and thus
the amount of echo returned to the transducer) depends
primarily on the ratio of the acoustic impedances at the
interface between two tissues and the angle at which
the ultrasound beam hits the structure of interest (angle
of incidence equals angle of reflection). An improper
transducer orientation, steering angle (i.e., the angle at
which ultrasound waves are emitted from the trans-
ducer) or rotation of muscle fascicles during contrac-
tion may therefore lead to echoes not being detected by
the transducer.
Because of their high water content, skeletal
muscles appear as mostly hypo-echoic (i.e., dark) struc-
tures (Ihnatsenka and Boezaart 2010). In the transverse
plane, reflections of comparatively hyper-echoic perimy-
sial connective tissue give muscles a speckled appear-
ance. In the longitudinal plane, by contrast, reflections of
perimysial connective tissue appear as white lines, thus
revealing the architecture of skeletal muscles. Here,
careful manipulation is required to align the transducer
2493
to the muscle fascicle plane and optimize the echogenic-
ity of muscle fascicles (Lichtwark 2017).
The length of the muscle fascicles (Lf) and the penna-
tion angle (PA), which is the angle of insertion of the fas-
cicles into the muscle’s aponeurosis, represent the main
structural parameters that can be detected via the conven-
tional B-mode ultrasound modality (Narici et al. 2016). The
analysis of muscle architecture features can be carried out
with an image analysis software (e.g., ImageJ, National Insti-
tutes of Health, Bethesda, MD, USA) on previously col-
lected ultrasound scans: Briefly, after identification of the
superficial and deep aponeuroses from which the muscle fas-
cicles originate and insert into, the lengths of one or several
clearly visible muscle fascicles can be traced. Similarly, PA
values can be measured by assessing the angles enclosed by
fascicles and the aponeurosis into which they insert. For a
detailed description of such analyses, please refer to the pio-
neering works of Rutherford and Jones (1992), Narici et al.
(1996a) and Fukunaga et al. (1997).
B-Mode ultrasound offers the unique advantage of
facilitating examinations of contracting muscle. During con-
traction, muscle fascicles may dynamically change length
and rotate about their origin, resulting in changes in both Lf
and PA. To facilitate the (otherwise tedious and time-
consuming) analysis of image sequences acquired during
contraction, computer (mostly optical flow) algorithms
may be used for automated fascicle tracking (Cronin
et al. 2011; Farris and Lichtwark 2016). Another archi-
tectural parameter that is often assessed is muscle thick-
ness (MT), which is regarded as an easily assessable
measure of muscle size. In longitudinal ultrasound scans,
the MT of a single muscle is defined as the perpendicular
distance between the superficial and deep aponeurosis
(Franchi et al. 2017; Narici 1999). Figure 1A is a repre-
sentative ultrasound image.
Potential and limitations
The use of ultrasound for the quantification of skele-
tal muscle size and architecture has risen exponentially in
the last 25 y. The main reasons for this are that ultrasound
allows capture of real-time images and represents a non-
invasive technique that is both cheaper than magnetic res-
onance imaging (MRI) and safer than other techniques
involving ionizing radiation, such as dual-energy X-ray
absorptiometry (DXA) or computed tomography (CT).
Further, although MRI is regarded as the gold standard
for clinical imaging and research, ultrasound is easier to
implement and virtually unrestricted by exclusion criteria.
It allows muscle architecture images that have superior
temporal and spatial resolution compared with MRI
images to be obtained in real time (Lopata et al. 2010).
These features have made B-mode ultrasound the pre-
ferred method for studying dynamic muscle architecture
changes, for instance, during peripheral nerve stimulation
2494 Ultrasound in Medicine & Biology Volume 44, Number 12, 2018

Fig. 1. Longitudinal B-mode ultrasound scans of the vastus lateralis, acquired with (A) a 5-cm transducer and (B) a
10-cm transducer, illustrating fascicle length (Lf), pennation angle (PA) and muscle thickness (MT). A greater section of
total muscle fascicle length must be estimated through linear extrapolation of visible segments when vastus lateralis
scans are acquired with a 5-cm transducer.

(Mayfield et al. 2015), voluntary contraction and passive
joint rotations (Herbert and Gandevia 1995; Narici et al.
1996a,). Although ultrasound is easily applicable, meas-
urements of muscle architecture are operator dependent,
which has led several researchers to question the reliabil-
ity of the technique. However, several research groups
have found that (as for most technical equipment), with
appropriate operator training, measures can be highly
reproducible (Kwah et al. 2013), as reflected by intra-class
correlation coefficients of >0.99 (Aeles et al. 2017; Ver-
nillo et al. 2017).
One key limitation of standard B-mode ultrasound
is its relatively small field of view, which is determined
by the size of the (linear array) transducer. The field of
view typically ranges from 4 to 6 cm, although it may be
as large as 10 cm in some instances. Because of this limi-
tation, it is important to mention that different muscles
have distinct structural arrangements (Friederich and
Brand 1990; Wickiewicz et al. 1983). From data
obtained from cadaveric studies, the two most com-
monly investigated muscles in humans, the gastrocne-
mius medialis (GM) and vastus lateralis (VL), present an
average Lf (as measured at rest and with the ankle and
knee held in a neutral and fully extended position,
respectively) between »3.5 and 5 cm for GM and
between »6.5 and 8 cm for VL (Friederich and Brand
1990; Huijing 1985; Wickiewicz et al. 1983). In other
muscles, such as the human sartorius muscle, however,
Lf may exceed 20 cm (Heron and Richmond 1993). Tak-
ing into consideration that cadaver data can be affected
by fiber shrinkage, Lf values could be expected to be
even higher in in vivo examinations. It follows that Lf
may be easily measured in some muscles using a stan-
dard ultrasound transducer, whereas only estimates
(based on the extrapolation of visible segments) of Lf
may be possible in other muscles. Consequently, muscle
Lf measurements from more proximal leg muscles are
inherently subjected to greater measurement inaccura-
cies (Fig. 1).
One way to overcome the limitation of a small
field of view is to choose a longer transducer: Commer-
cially available transducers are typically 4 to 5 cm long
and, therefore, ideal for the study of muscles with
shorter Lf (e.g., triceps surae muscle group). Longer
transducers (from 6 to 10 cm) are more suitable for
muscles that have longer Lf (e.g., quadriceps femoris
muscle group), but limited in their temporal resolution,
because the acquisition frame rate decreases with a
larger field of view. Although this limitation may have
a negligible influence on scans performed at rest or dur-
ing slow passive joint rotations, it may impede the
study of muscular contractions involved in faster move-
ments. Further, body surface contours are rarely
straight, so the usage of longer transducers may lead to
uneven compression of underlying tissues and, conse-
quently, muscle deformations (Wakeling et al. 2013).
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al.
To obtain larger-field-of-view images without access to
a longer transducer, some groups have proposed use of
two 5-cm linear transducers placed in series (Brennan
et al. 2017; Herbert et al. 2011) to cover a total length
of approximately 1112 cm (1- to 2-cm gap in between
the probes). Brennan et al. (2017) found that this “dual
array” setup allows more precise estimations of abso-
lute fascicle length compared with scans obtained using
a single transducer. This was because this method
allowed the VL fascicles to be displayed almost in their
entirety: however, as the two 5-cm transducers would
have to be paired on a stiff frame strapped to the leg,
this strategy would not avoid tissue deformation caused
by compression.
In cases in which neither one longer linear trans-
ducer nor two shorter ones are available, Lf can be esti-
mated using extrapolation methods. One approach,
proposed by Blazevich et al. (2006), relies on the simpli-
fying assumption that the shape of a portion of pennate
muscle resembles a parallelogram (Haxton 1944; Narici
1999). Here, Lf is given by
 1
Lf ¼ sinðg þ 90B Þ ¢ MT ¢ sin 180B  ðg þ 180B PAÞ
where g is the angle formed by the deep and superficial
aponeuroses (Blazevich et al. 2006). Although the angle
between the two aponeuroses is taken into account, Lf
predictions made using this equation are still highly
influenced by MT, which is not constant along the
muscle’s length and could follow a non-linear pattern
outside the visible field of view (i.e., the MT measured
in the center of the muscle belly is typically greater than
those measured in other muscle areas). Further, fascicles
are often curved, which is not accounted for in such cal-
culations (i.e., the equation assumes that fascicles are
represented accurately by a straight line) as well as for
regional architectural differences within the same mus-
cle. Operators and researchers should be aware of these
potential limitations, especially when examining
muscles with longer fascicles such as the knee extensor
and hamstring muscles. As an example, a recent study
by Freitas et al. (2018), in which a 6-cm transducer was
used, compared Lf calculations from the biceps femoris
long head that relied on either full (length of field of
view: 6 cm) or half (field of view: 3 cm) images and
used the above equation. The results were substantially
different (9.9910.47 cm vs. 12.0313.89 cm, respec-
tively), which underlines the influence of transducer
length on Lf measures.
Regional differences in muscle architecture have
been observed not only between muscles with different
functional roles, but also between heads of synergistic
muscle groups and even within the same muscle
(Blazevich et al. 2006). Recent data also suggest that
2495
training-induced architectural adaptations (changes in Lf
and PA) are specific to the muscle region examined
(Ema et al. 2013). The standardization of scanning sites
is therefore crucial when longitudinal muscle architec-
ture changes are assessed, as the hypertrophic responses
to different training stimuli may differ throughout the
muscle’s length (Franchi et al. 2014; Narici et al. 1989;
Seger et al. 1998), which complicates the deduction of
total muscle growth from changes in MT as observed in
single muscle regions. Indeed, a recent study reported
good agreement between the training-induced changes
in MT and muscle cross-sectional area (as assessed by
MRI at the same muscle site) (Franchi et al. 2017). How-
ever, changes in MT were not statistically related to
changes in muscle volume: although this was not surpris-
ing, as mathematically one would not expect a volumet-
ric (3-D) measure to increase in direct proportion to a
linear measure, the discrepancy between MT and volume
changes suggests that regional hypertrophic responses
(or shape remodeling along the whole muscle lengths)
may have occurred and ultimately influenced the correla-
tion. Changes in total muscle size should, therefore, be
assessed by other imaging techniques, such as MRI, or
scans should be obtained at multiple sites of interest.
The extended-field-of-view ultrasound technique (see
next section) may offer further advantages in providing
insights at a whole-muscle level.
EXTENDED-FIELD-OF-VIEW ULTRASOUND
IMAGING
Principles
The EFOV technique was introduced at the end of
the 1990s (Weng et al. 1997) to facilitate the study of
longer anatomic structures. This technique relies on tex-
ture mapping algorithms to merge sequences of images
collected during real-time scanning to reconstruct large
composite images. As the transducer is moved along the
skin surface and along a plane of interest, successive
frames are recorded and stitched together to obtain a
panoramic image. The features of each frame serve as a
reference to match the subsequent frame and autocorre-
lation algorithms, and an image processor creates a pan-
oramic image (Weng et al. 1997). The software
continuously compares the features of the current frame
(the real-time portion of the image) with those of the pre-
ceding frame (the static portion of the image), which
allows for accurate positioning of the current frame with
respect to the previous frames. Because of the absence
of respiratory movements and pulsatility of large vessels,
the musculoskeletal system is particularly well suited for
EFOV imaging (Lin et al. 1999; Weng et al. 1997).
Indeed, errors of EFOV distance measurements performed
2496 Ultrasound in Medicine & Biology
on phantoms up to 70 cm in length were less than 5% (For-
nage 2000; Weng et al. 1997).
Potential and limitations
The EFOV technique can be used for both trans-
verse and longitudinal scans of skeletal muscle.
Although transverse scans allow images of anatomic
cross-sectional areas of muscles or muscle groups of
interest to be obtained, longitudinal scans are acquired
for the assessment of muscle architecture.
Longitudinal EFOV scans, compared with single B-
mode images, have the obvious advantage of providing a
panoramic view of the whole fascicle length and thus
facilitating the assessment of long fascicles (Fig. 2). It is
important to note that muscles typically do not present a
homogeneous architectural arrangement along their whole
length (Blazevich et al. 2006), and the fascicle pattern can
change between different muscle sites (e.g., from mid- to
distal portion). Although EFOV ultrasound allows visuali-
zation of such architectural heterogeneity, the technique is
susceptible to bias related to transducer misalignments.
This is because it may be challenging to satisfy the funda-
mental requirement of keeping the transducer parallel to
the fascicle plane when covering large areas of interest,
and it is known that the orientation and positioning of the
ultrasound transducer are crucial for muscle architectural
measurements (Benard et al. 2009; Klimstra et al. 2007).
In the light of this challenge, several studies (Adkins et al.
2017; Noorkoiv et al. 2010; Seymore et al. 2017; Simpson
et al. 2017) have examined the reliability of the EFOV
technique. In a pioneering study, Noorkoiv et al. (2010)
reported high intra- and inter-experimenter reliability for
VL Lf as obtained from scans of »18-cm length (coincid-
ing with approximately 50% of VL muscle length in
adults). It should, however, be noted that the muscle seg-
ment studied in this paper did not extend to regions close
to either the proximal or distal myotendinous junction
(MTJ)—areas that are of particular scientific and clinical
interest and are often distinguished by a fascicle arrange-
ment very different from that seen in the center of the
muscle belly. In the long head of the biceps femoris mus-
cle, Seymore et al. (2017) recently investigated muscle
architecture over its whole length and similarly found
excellent measurement reproducibility (Lf: intra-class cor-
relation coefficient [ICC] = 0.99, standard error of mea-
surement [SEM] = 0.11 cm; PA: ICC = 0.88, SEM = 1.1˚).
However, there are inherent difficulties in scanning large
regions with variable architectural features. Substantial
examiner training is imperative to warrant adequate mea-
surement reproducibility.
When used for scans in the transverse plane, EFOV
may also represent a useful alternative to MRI in the
assessment of muscle anatomic cross-sectional area
(Ahtiainen et al. 2010; Noorkoiv et al. 2010) and muscle
Volume 44, Number 12, 2018
volume (Infantolino et al. 2007). Potential limitations lie
in the use of linear transducers for the assessment of
curved surfaces and the pressure exerted by the operator
on the skin and underlying muscle tissue, which may
influence measurement results. Nonetheless, although
MRI systematically produced larger anatomic cross-sec-
tional area values than ultrasound, EFOV-based data
were in good agreement with MRI (Ahtiainen et al. 2010).
Two further limitations of the technique are that
EFOV cannot provide insight into the deformation of
muscle during contraction and that it yields (just as stan-
dard B-mode ultrasound) only 2-D images. To overcome
the latter, 3-D ultrasound imaging, which is covered in
the next section, has been developed.
3-D ULTRASOUND IMAGING
Principles
Although conventional B-mode ultrasound imaging
is useful for measuring static and dynamic muscle archi-
tecture changes in a single plane, its limited field of view
does not always allow the entire muscle fascicle length
to be imaged. Similarly, muscle belly length, aponeuro-
sis length and muscle volume cannot be quantified.
To overcome these limitations, freehand 3-D ultrasound
(3-DUS) was developed, and so far this technique has
been implemented to measure in vivo human muscle vol-
ume and muscle belly length (Barber et al. 2009; Raiteri
et al. 2016), fascicle length (Hug et al. 2015; Raiteri
et al. 2016), proximal-to-distal pennation angles
(Hiblar et al. 2003) and 3-D muscle fascicle architecture
(Rana et al. 2013). More recently, central aponeurosis
widths and lengths of the human tibialis anterior muscle
have also been quantified (Raiteri et al. 2016). Another
benefit of this technique is that the distance between
ultrasound images can be minimized by increasing the
scan time, which improves the spatial resolution of the
reconstructed 3-D images. Lastly, compared with EFOV
technique, during 3-D ultrasound, images are registered
and not acquired on a pixel-pattern basis and thus are
independent of artifactual features.
Briefly, 3-DUS combines conventional B-mode
ultrasound imaging with synchronous tracking of the
ultrasound transducer’s position and orientation (through
electromagnetic or optical methods) to form a 3-D data
set that describes the muscle volume in the global coor-
dinate system. A known calibration of the ultrasound
image position relative to the transducer must be applied
so that images can be correctly projected into the global
coordinate system and precise voxel information can be
generated (Treece et al. 2003). Muscle slices through
this volume can then be reconstructed and viewed,
allowing muscle architecture images that are not
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al. 2497

Fig. 2. Images of (A) gastrocnemius medialis and (B) vastus lateralis muscles obtained by extended-field-of-view ultra-
sound technique. Whereas only about 55% of the full length of the gastrocnemius medialis is displayed in the image
(from the distal myotendinous junction upward), the vastus lateralis is shown in almost its entire length, from the distal
myotendinous junction at the patella to its proximal portion, almost at the great trochanter spot.

accessible with standard B-mode ultrasound to be ana-
lyzed. To determine muscle volume, the transducer must
be swept along the length of the muscle, and the muscle
borders of a sufficient number of transverse plane ultra-
sound images need to be delineated to represent the mus-
cle belly shape accurately. For larger muscles, multiple
overlapping ultrasound sweeps along the length of the
muscle are required to image the muscle borders ade-
quately, so that the muscle cross section can be imaged
in its entirety (Barber et al. 2009). In this case, it is rec-
ommended that a water bath is used (Barber et al. 2009),
instead of an ultrasound gel pad (Raiteri et al. 2016), to
limit the influence of transducer pressure changes on
image processing (Fig. 3).
Potential and limitations
Muscles with highly visible borders along their
length and widths smaller than the transducer’s field of
view are ideal for 3-DUS imaging as they can be imaged
in their entirety with a single sweep of the ultrasound
transducer, which improves the quality of the scans
and reduces errors associated with the subjective seg-
mentation process. A modified version of sweeping the
muscle, using a 2-cm-thick echogenic ultrasound gel pad
(Aquaflex, Parker Laboratories, Fairfield, NJ, USA),
rather than a water bath, has previously been found to be
an effective method to image the entire muscle belly dur-
ing single-sweep scans (Raiteri et al. 2016). Reducing
the frame rate (to 1015 Hz) to improve image quality
(i.e., through a higher line density) is recommended to
improve the muscle segmentations, even if this extends
the scan time slightly.
A greater number of segmentations will generally
need to be performed at the ends of the muscle because
of the more irregular and dissimilar cross sections,
whereas segmentations in the midbelly can be reduced if
the cross sections are relatively uniform to decrease
post-processing time (typically 1530 min). Aponeuro-
sis widths and lengths can be best visualized in bi-pen-
nate muscles as the central aponeurosis is relatively easy
to identify (Raiteri et al. 2016).
One of the major benefits of 3-DUS is that the 3-D
muscle and central aponeurosis deformations during
fixed-end contractions can be quantified in multiple
planes. Muscle cross-sectional area, thickness and width
changes in the human tibialis anterior have previously
been determined up to 50% of maximum voluntary
isometric contraction torque (Raiteri et al. 2016), after
muscle and aponeurosis segmentations, the use of a
shape-based paradigm (Treece et al. 2000) to interpolate
a surface through the segmented muscle slices and to
turn the muscle cross-sections into a 3-D triangle based
 2498
Ultrasound in Medicine & Biology
Volume 44, Number 12, 2018
Fig. 3. Three-dimensional ultrasound image capture. (A) Transverse scan of the mid-portion of the human tibialis anterior muscle. (B) Position of the scan along the
whole muscle volume. (C) Reconstruction of the sagittal muscle plane.
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al.
model (Treece et al. 1999), and the implementation of a
weighted principal component analysis to define the
axes of the aponeurosis. The method of re-slicing the
muscle in the transverse plane along the longitudinal
axis of its central aponeurosis enables accurate measures
of muscle thickness that might not be possible using a
fixed image plane as in conventional B-mode ultrasound
imaging, where the degree of measurement error is
clearly influenced by the transducer alignment
(Bolsterlee et al. 2016). Transducer fixation during con-
ventional ultrasound imaging may also cause greater
muscle compression than 3-DUS, which may influence
muscle deformations (Wakeling et al. 2013). Lastly,
because the exact endpoints of the muscle or aponeurosis
can be determined with 3-DUS, it is likely that global
strains can be determined more precisely than with a
conventional B-mode ultrasound approach.
A key limitation to 3-DUS is that high contraction
intensities and dynamic muscle architecture changes
cannot be studied because of the time it takes to perform
a scan. Even during fixed-end contractions, joint rota-
tions must be minimized so that length changes are not
overestimated. Operator training is also required to
ensure that the muscle borders can be completely cap-
tured and to ensure the transducer’s orientation and
speed can be maintained during a scan, so that transverse
images do not overlap and the distance between frames
can be kept small. Despite these technical difficulties
and the need for further hardware and software develop-
ment, it is clear that 3-DUS can provide useful informa-
tion regarding how a muscle and its aponeurosis deform
in multiple planes simultaneously during contractions at
different forces and muscle lengths. This information
will certainly be useful in understanding the role of mus-
cle bulging and aponeurosis deformations during con-
traction and the implications for muscle performance,
muscle injury risk and rehabilitation.
DIFFUSION TENSOR IMAGING
Principles
The basic principles of diffusion tensor MRI (DTI)
were first introduced in the mid-1980s (Le Bihan and
Breton 1985; Taylor and Bushell 1985). The rather com-
plex physical concepts of DTI have been described
extensively in the literature. For a detailed, yet readily
understandable introduction, readers are referred to the
explanations by Mori and Zhang (2006). In brief, the
technique relies on the fact that the Brownian motion of
(mostly water) molecules in tissues is not entirely free,
but interrupted by the collisions with anatomic obstacles,
such as particles within the cell, membranes or other
macromolecules. Water diffusion in biological tissues is
anisotropic in that it occurs rapidly in the direction
2499
aligned in parallel with internal structures, whereas it is
hindered perpendicularly to the preferred direction. By
studying the anisotropy of water diffusion, it is, there-
fore, possible to probe tissue structure in 3-D space.
Magnetic resonance imaging pulse sequences, such as
the T2-weighted single-shot echo-planar imaging sequence
most commonly used for DTI (Nana et al. 2008), can be
designed to be sensitive to water diffusion by applying two
subsequent gradient pulses (also known as the motion-
probing gradients). The first of these pulses induces a phase
shift in proton precession, whereas the second (typically
2050 ms after the first) aims to reverse it. The detection
of water diffusion relies on the fact that after the adminis-
tration of these two pulses, static particles will experience
no net phase shift, while the second pulse will fail to per-
fectly restore the original proton spins in moving particles.
Thus, particles that have moved will end up with a phase
shift that leads to an exponential decay in signal intensity,
where the amount of decay is proportional to the diffusion
coefficient of the water molecules and the degree of diffu-
sion weighting of the diffusion-gradient pulses.
The degree of diffusion weighting depends on the
strength (height) and duration (breadth) of these diffusion
gradient pulses, as well as the time between them, which
determine their b value. Higher b values imply stronger
diffusion weighting that probes lower diffusion values
and results in overall darker images, because tissues lose
greater signal intensity at larger b values. Isotropic diffu-
sion, which is not dependent on direction, can be experi-
mentally determined by application of the diffusion
sensitizing gradient along any direction. As mentioned
before, in tissues with a microarchitecture that is oriented
preferentially in one predominant direction, the diffusion
of the water molecules is greater in one direction. For
instance, in the axons of white matter of the brain or mus-
cle fascicles, diffusion occurs to a greater extent along the
axons or muscle fibers compared with the other (cross-
sectional) directions. Such anisotropy of diffusion is char-
acterized by the diffusion tensor. DTI allows this tensor to
be quantified and uses sequences with diffusion gradient
pulses applied in different directions. A DTI sequence
acquires a series of images with the same b value, with
diffusion gradients applied in different directions, to cre-
ate tensor maps of water diffusion, that is, the diffusion
tensor images. The 3 £ 3 diffusion tensor has six indepen-
dent values, so a minimum of six non-collinear diffusion
gradients are required to solve for the six unknowns
(Basser et al. 1994). This allows for the determination of
the 3 £ 3 diffusion tensor in the image frame of reference
and will have both diagonal and off-diagonal terms.
Matrix diagonalization aligns this tensor with the main
axes of diffusion and yields the respective eigenvectors
and associated eigenvalues. In skeletal muscles, the eigen-
vector corresponding to the lead or primary eigenvalue of
2500 Ultrasound in Medicine & Biology
the diagonalized tensor has been reported to correspond
to the fiber direction (Damon et al. 2002). The average of
all three eigenvalues is commonly referred to as mean dif-
fusivity (MD); a theoretical value of 0 would reflect com-
plete absence of diffusion and, thus, a lack of Brownian
motion of molecules. Another frequently reported metric
is the fractional anisotropy (FA), which measures the
degree of directionality of water diffusion on a voxel-by-
voxel basis. FA ranges from 0, which corresponds to a
sphere or isotropic diffusion of water molecules in a
beaker, to 1, which corresponds to an infinitesimally thin
tube as in a muscle fiber or neuron, while typically inter-
mediate non-zero values correspond to an ellipsoidal shape.
Once the diffusion tensor has been determined,
fibers can be tracked (Damon et al. 2011). After a seed
point is defined, often manually, an algorithm looks for
adjacent voxels whose main diffusion direction is con-
tinuous with the previous one. Once these voxels are
linked, the paths of corresponding anatomic structures,
such as neurons and muscle fascicles, can be recon-
structed (Budzik et al. 2007). Tracking stops when a cut-
off value (e.g., concerning the minimal or maximal FA
value or the angular change of the fiber direction per
integration step) is reached.
Potential and limitations
Diffusion tensor visualization and associated fiber trac-
tography have been increasingly used for the study of skele-
tal muscle architecture at rest (Damon et al. 2017).
Feasibility studies have confirmed the reliability of DTI in
determining fiber lengths, pennation angles and physiologic
cross-sectional areas (Okamoto et al. 2010; Sinha et al.
2011a). Sinha et al. (2011a, 2011b) further documented the
effects of ankle joint angle changes on DTI-based measures
of human soleus muscle architecture. Damon et al. (2012)
used DTI to accurately estimate muscle fiber curvature,
which is relevant for the prediction of strain patterns during
contraction. More recently, DTI was applied in a cross-sec-
tional study to document age-associated differences in tri-
ceps surae architecture (Sinha et al. 2015). The main
strength of DTI is that it represents the only currently avail-
able technique to study muscle architecture non-invasively
in three dimensions over a large area of interest, which is
not limited by tissue depth. In fact, the wealth of architec-
tural information provided by DTI cannot even be accounted
for by most current biomechanical models of skeletal mus-
cle, which are frequently simplistic and depict muscle archi-
tecture as a uniform structure of linear muscle fascicles
arranged in parallel.
Although DTI represents a superior technique for
studying muscle architecture, the analysis of water diffu-
sion may also provide information about tissue micro-
structure that is not visible at the macroscopic scale. For
instance, changes in fiber diameter resulting from
Volume 44, Number 12, 2018
neurotomy have been found to be reflected by increased
FA values (Saotome et al. 2006; Zhang et al. 2008). Con-
versely, decreased FA values in combination with an
overall increased diffusivity (greater MD) are reportedly
indicative of sterile muscle trauma (Esposito et al. 2013;
Fan and Does 2008). Esposito et al. (2013) reported that,
jointly, FA and standard T2 values correlate well with
immunologic markers of muscle damage and repair. In
children with Duchenne muscle dystrophy, FA was
found to be strongly correlated with muscle fat content
and negatively correlated with muscle strength
(Ponrartana et al. 2015). Exercise-associated muscle
damage has been reported to result in increases in both
MD and FA (McMillan et al. 2011; Yanagisawa et al.
2010; Zaraiskaya et al. 2006). To summarize, DTI may
serve as a valuable biomarker to monitor and quantify
changes in muscle status and structure associated with
injury, disease or physiologic processes (for an overview
of recent studies that have implemented DTI in both
basic and applied clinical research, readers are referred
to the extensive review by Oudeman et al. [2016])
(Fig. 4). In this regard, the full potential of the technique
is only starting to be appreciated.
The main drawbacks of DTI are the comparably long
scanning times (e.g., »15 min for acquisition of all
required image stacks to analyze the tibialis anterior mus-
cle [Heemskerk et al. 2010]) and associated examination
costs, as well as the impossibility of acquiring images of
muscles in the contracted state. Further, DTI and fiber
tractography involve rather complex image post-process-
ing routines that require substantial examiner training and
preclude the obtaining of “live” results. As in all MRI
experiments, imaging artifacts may arise from subject
motion. Additional artifacts frequently encountered in
DTI include susceptibility-induced deformations, eddy
current distortions and chemical shift artifacts. The effects
of these deformations can be limited through the registra-
tion of DTI images with high-resolution, non-diffusion-
weighted (b0) images (Heemskerk et al. 2009). Finally, it
must be mentioned that the placement of seed points and
definition of stop criteria for fiber tracking are subjective
and, therefore, a potential source of bias (e.g., changes in
stop criteria may drastically change measures of fiber
length because in their end regions, fascicles tend to coa-
lesce with aponeuroses).
PERSPECTIVE
This review presents various imaging-based
techniques that can be applied to the in vivo study of
skeletal muscle architecture and discusses their main
strengths and limitations. Conventional B-mode
ultrasound imaging has been most commonly used. It
is cost effective, easily applicable and best suited for
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al.
Fig. 4. Diffusion tensor imaging-based fiber reconstructions of the human tibialis anterior and gastrocnemius (A) and the external anal sphincter muscle (B). Part A is
reprinted from Oudeman et al. (2016) with permission.

2501
2502 Ultrasound in Medicine & Biology
the study of the dynamic changes of muscle architec-
ture during contraction. Its main limitation is the lim-
ited field of view that may impede the visualization
of longer fascicles. Using image mapping algorithms
to merge sequences of scans into composite images,
EFOV ultrasound overcomes this limitation in pas-
sive or constant force conditions, although care must
be taken to avoid bias related to transducer misalign-
ments when covering larger areas of interest. 3-DUS
further expands the potential of EFOV ultrasound
in that it acquires information in three dimensions
and, thus, facilitates the reconstruction of arbitrary
sectional images. Image acquisition (and reconstruc-
tion) is more complex and time consuming, thus pre-
cluding the study of dynamic architecture changes.
DTI may provide exquisite 3-D images of muscle
architecture and cover large areas of interest and, as
opposed to ultrasound-based techniques, is not lim-
ited by tissue depth. In addition, the study of water
diffusion may allow for conclusions about muscle tis-
sue microstructure that may be indicative of the mus-
cle status in health, injury or disease. At the same
time, it represents the most complex, time consuming
and expensive of all currently available techniques
and cannot be applied to the study of muscle
contraction. Hence, the appropriate methodological
approach must be carefully selected in dependency of
the scientific questions or clinical demands to be met.
Acknowledgments—We thank Professor Martijn Froeling for kindly
granting us permission to reprint the diffusion tensor image of the
lower leg (Fig. 4A).
CONFLICT OF INTEREST DISCLOSURE
The authors declare that there was no conflict of interest in the
writing of this article.
REFERENCES
Adkins AN, Franks PW, Murray WM. Demonstration of extended
field-of-view ultrasound’s potential to increase the pool of muscles
for which in vivo fascicle length is measurable. J Biomech
2017;63:179–185.
Aeles J, Lichtwark GA, Lenchant S, Vanlommel L, Delabastita T, Van-
wanseele B. Information from dynamic length changes improves
reliability of static ultrasound fascicle length measurements. PeerJ
2017;5:e4164.
Ahtiainen JP, Hoffren M, Hulmi JJ, Pietikainen M, Mero AA, Avela J,
Hakkinen K. Panoramic ultrasonography is a valid method to mea-
sure changes in skeletal muscle cross-sectional area. Eur J Appl
Physiol 2010;108:273–279.
Barber L, Barrett R, Lichtwark G. Validation of a freehand 3 D ultra-
sound system for morphological measures of the medial gastrocne-
mius muscle. J Biomech 2009;42:1313–1319.
Basser PJ, Mattiello J, LeBihan D. Estimation of the effective self-dif-
fusion tensor from the NMR spin echo. J Magn Reson B
1994;103:247–254.
Benard MR, Becher JG, Harlaar J, Huijing PA, Jaspers RT. Anatomical
information is needed in ultrasound imaging of muscle to avoid
Volume 44, Number 12, 2018
potentially substantial errors in measurement of muscle geometry.
Muscle Nerve 2009;39:652–665.
Blazevich AJ, Gill ND, Zhou S. Intra- and intermuscular variation in
human quadriceps femoris architecture assessed in vivo. J Anat
2006;209:289–310.
Bolsterlee B, Gandevia SC, Herbert RD. Effect of transducer orienta-
tion on errors in ultrasound image-based measurements of human
medial gastrocnemius muscle fascicle length and pennation. PLoS
One 2016;11 e0157273.
Brennan SF, Cresswell AG, Farris DJ, Lichtwark GA. In vivo fascicle
length measurements via B-mode ultrasound imaging with single
vs dual transducer arrangements. J Biomech 2017;64:240–244.
Budzik JF, Le Thuc V, Demondion X, Morel M, Chechin D, Cotten A.
In vivo MR tractography of thigh muscles using diffusion imaging:
Initial results. Eur Radiol 2007;17:3079–3085.
Chan VWS, Abbas S, Brull R, Morrigl B, Perlas A. Ultrasound imaging
for regional anesthesia. A practical guide. 2nd ed Toronto, ON:
Toronto Publishing; 2008.
Cronin NJ, Carty CP, Barrett RS, Lichtwark G. Automatic tracking of
medial gastrocnemius fascicle length during human locomotion. J
Appl Physiol (1985) 2011;111:1491–1496.
Damon BM, Ding Z, Anderson AW, Freyer AS, Gore JC. Validation of
diffusion tensor MRI-based muscle fiber tracking. Magn Reson
Med 2002;48:97–104.
Damon BM, Buck AK, Ding Z. Diffusion-Tensor MRI based skeletal
muscle fiber tracking. Imaging Med 2011;3:675–687.
Damon BM, Heemskerk AM, Ding Z. Polynomial fitting of DT-MRI
fiber tracts allows accurate estimation of muscle architectural
parameters. Magn Reson Imaging 2012;30:589–600.
Damon BM, Froeling M, Buck AK, Oudeman J, Ding Z, Nederveen AJ,
Bush EC, Strijkers GJ. Skeletal muscle diffusion tensor-MRI fiber track-
ing: Rationale, data acquisition and analysis methods, applications and
future directions. NMR Biomed 2017;30:e3563.
Ema R, Wakahara T, Miyamoto N, Kanehisa H, Kawakami Y. Inhomo-
geneous architectural changes of the quadriceps femoris induced by
resistance training. Eur J Appl Physiol 2013;113:2691–2703.
Esposito A, Campana L, Palmisano A, De Cobelli F, Canu T, Santar-
ella F, Colantoni C, Monno A, Vezzoli M, Pezzetti G, Manfredi
AA, Rovere-Querini P, Del Maschio A. Magnetic resonance
imaging at 7 T reveals common events in age-related sarcopenia
and in the homeostatic response to muscle sterile injury. PLoS
One 2013;8:e59308.
Fan RH, Does MD. Compartmental relaxation and diffusion tensor
imaging measurements in vivo in lambda-carrageenan-induced
edema in rat skeletal muscle. NMR Biomed 2008;21:566–573.
Farris DJ, Lichtwark GA. UltraTrack: Software for semi-automated
tracking of muscle fascicles in sequences of B-mode ultrasound
images. Comput Methods Programs Biomed 2016;128:111–118.
Fornage BD. The case for ultrasound of muscles and tendons. Semin
Musculoskeletal Radiol 2000;4:375–391.
Franchi MV, Atherton PJ, Reeves ND, Fluck M, Williams J, Mitchell
WK, Selby A, Beltran Valls RM, Narici MV. Architectural, func-
tional and molecular responses to concentric and eccentric loading in
human skeletal muscle. Acta Physiol (Oxford) 2014;210:642–654.
Franchi MV, Longo S, Mallinson J, Quinlan JI, Taylor T, Greenhaff
PL, Narici MV. Muscle thickness correlates to muscle cross-sec-
tional area in the assessment of strength training-induced hypertro-
phy. Scand J Med Sci Sports 2017;28:846–853.
Freitas SR, Marmeleira J, Valamatos MJ, Blazevich A, Mil-Homens P.
Ultrasonographic measurement of the biceps femoris long-head
muscle architecture. J Ultrasound Med 2018;37:977–986.
Friederich JA, Brand RA. Muscle fiber architecture in the human lower
limb. J Biomech 1990;23:91–95.
Fukunaga T, Ichinose Y, Ito M, Kawakami Y, Fukashiro S. Determina-
tion of fascicle length and pennation in a contracting human muscle
in vivo. J Appl Physiol (1985) 1997;82:354–358.
Haxton HA. Absolute muscle force in the ankle flexors of man. J Phys-
iol 1944;103:267–273.
Heemskerk AM, Sinha TK, Wilson KJ, Ding Z, Damon BM. Quantita-
tive assessment of DTI-based muscle fiber tracking and optimal
tracking parameters. Magn Reson Med 2009;61:467–472.
 Muscle Architecture Assessment: Strengths, Shortcomings and New Frontiers
M. V. FRANCHI et al.
Heemskerk AM, Sinha TK, Wilson KJ, Ding Z, Damon BM. Repeat-
ability of DTI-based skeletal muscle fiber tracking. NMR Biomed
2010;23:294–303.
Herbert RD, Gandevia SC. Changes in pennation with joint angle and
muscle torque: In vivo measurements in human brachialis muscle. J
Physiol 1995;484(Pt 2):523–532.
Herbert RD, Clarke J, Kwah LK, Diong J, Martin J, Clarke EC, Bilston
LE, Gandevia SC. In vivo passive mechanical behaviour of muscle
fascicles and tendons in human gastrocnemius muscle-tendon units.
J Physiol 2011;589:5257–5267.
Heron MI, Richmond FJ. In-series fiber architecture in long human
muscles. J Morphol 1993;216:35–45.
Hiblar T, Bolson EL, Hubka M, Sheehan FH, Kushmerick MJ.
Three dimensional ultrasound analysis of fascicle orientation in
human tibialis anterior muscle enables analysis of macroscopic
torque at the cellular level. Adv Exp Med Biol 2003;538:635–
644 discussion 645.
Hug F, Goupille C, Baum D, Raiteri BJ, Hodges PW, Tucker K. Nature
of the coupling between neural drive and force-generating capacity in
the human quadriceps muscle. Proc Biol Sci 2015;282 20151908.
Huijing PA. Architecture of the human gastrocnemius muscle and some
functional consequences. Acta Anat (Basel) 1985;123:101–107.
Ihnatsenka B, Boezaart AP. Ultrasound: Basic understanding and learn-
ing the language. Int J Shoulder Surg 2010;4:55–62.
Infantolino BW, Gales DJ, Winter SL, Challis JH. The validity of
ultrasound estimation of muscle volumes. J Appl Biomech
2007;23:213–217.
Klimstra M, Dowling J, Durkin JL, MacDonald M. The effect of ultra-
sound probe orientation on muscle architecture measurement. J
Electromyogr Kinesiol 2007;17:504–514.
Kwah LK, Pinto RZ, Diong J, Herbert RD. Reliability and validity of
ultrasound measurements of muscle fascicle length and pennation in
humans: A systematic review. J Appl Physiol 2013;114:761–769.
Le Bihan D, Breton E. Imagerie de diffusion in-vivo par resonance
magnetique nucleaire. C R Acad Sci 1985;93:27–34.
Lichtwark G. Ultrasound technology for examining the mechanics of
the muscle, tendon, and ligament. In: M€uller B, Wolf SI,
Brueggemann GP, Deng Z, McIntosh A, Miller F, Selbie WS,
(eds). Handbook of human motion. Cham: Springer; 2017. p. 1–20.
Lieber RL, Friden J. Functional and clinical significance of skeletal
muscle architecture. Muscle Nerve 2000;23:1647–1666.
Lin EC, Middleton WD, Teefey SA. Extended field of view sonography
in musculoskeletal imaging. J. Ultrasound Med 1999;18:147–152.
Lopata RG, van Dijk JP, Pillen S, Nillesen MM, Maas H, Thijssen JM,
Stegeman DF, de Korte CL. Dynamic imaging of skeletal muscle
contraction in three orthogonal directions. J Appl Physiol (1985)
2010;109:906–915.
Mayfield DL, Lichtwark GA, Cronin NJ, Avela J, Cresswell AG. Doublet
potentiation in the triceps surae is limited by series compliance and
dynamic fascicle behavior. J Appl Physiol (1985) 2015;119:807–816.
McMillan AB, Shi D, Pratt SJ, Lovering RM. Diffusion tensor MRI to
assess damage in healthy and dystrophic skeletal muscle after
lengthening contractions. J Biomed Biotechnol 2011;2011 970726.
Mori S, Zhang J. Principles of diffusion tensor imaging and its applica-
tions to basic neuroscience research. Neuron 2006;51:527–539.
Nana R, Zhao T, Hu X. Single-shot multiecho parallel echo-planar
imaging (EPI) for diffusion tensor imaging (DTI) with improved
signal-to-noise ratio (SNR) and reduced distortion. Magn Reson
Med 2008;60:1512–1517.
Narici MV. Human skeletal muscle architecture studied in vivo by non-
invasive imaging techniques: Functional significance and applica-
tions. J Electromyogr Kinesiol 1999;9:97–103.
Narici MV, Roi GS, Landoni L, Minetti AE, Cerretelli P. Changes in
force, cross-sectional area and neural activation during strength
training and detraining of the human quadriceps. Eur J Appl Physiol
Occup Physiol 1989;59:310–319.
Narici MV, Binzoni T, Hiltbrand E, Fasel J, Terrier F, Cerretelli P. In vivo
human gastrocnemius architecture with changing joint angle at rest and
during graded isometric contraction. J. Physiol 1996;496(Pt 1):287–297.
Narici M, Franchi M, Maganaris C. Muscle structural assembly and
functional consequences. J Exp Biol 2016;219:276–284.
2503
Noorkoiv M, Nosaka K, Blazevich AJ. Assessment of quadriceps mus-
cle cross-sectional area by ultrasound extended-field-of-view imag-
ing. Eur J Appl Physiol Occup Physiol 2010;109:631–639.
Okamoto Y, Kunimatsu A, Kono T, Kujiraoka Y, Sonobe J, Minami M.
Gender differences in MR muscle tractography. Magn Reson Med
Sci 2010;9:111–118.
Oudeman J, Nederveen AJ, Strijkers GJ, Maas M, Luijten PR, Froeling
M. Techniques and applications of skeletal muscle diffusion tensor
imaging: A review. J Magn Reson Imaging 2016;43:773–788.
Ponrartana S, Ramos-Platt L, Wren TA, Hu HH, Perkins TG, Chia JM,
Gilsanz V. Effectiveness of diffusion tensor imaging in assessing
disease severity in Duchenne muscular dystrophy: Preliminary
study. Pediatr Radiol 2015;45:582–589.
Raiteri BJ, Cresswell AG, Lichtwark GA. Three-dimensional geometri-
cal changes of the human tibialis anterior muscle and its central
aponeurosis measured with three-dimensional ultrasound during
isometric contractions. PeerJ 2016;4:e2260.
Rana M, Hamarneh G, Wakeling JM. 3-D fascicle orientations in tri-
ceps surae. J Appl Physiol (1985) 2013;115:116–125.
Rutherford OM, Jones DA. Measurement of fibre pennation using ultra-
sound in the human quadriceps in vivo. Eur J Appl Physiol Occup
Physiol 1992;65:433–437.
Saotome T, Sekino M, Eto F, Ueno S. Evaluation of diffusional anisot-
ropy and microscopic structure in skeletal muscles using magnetic
resonance. Magn Reson Imaging 2006;24:19–25.
Seger JY, Arvidsson B, Thorstensson A. Specific effects of eccentric
and concentric training on muscle strength and morphology in
humans. Eur J Appl Physiol Occup Physiol 1998;79:49–57.
Selva Raj I, Bird SR, Shield AJ. Ultrasound measurements of skel-
etal muscle architecture are associated with strength and func-
tional capacity in older adults. Ultrasound Med Biol
2017;43:586–594.
Seymore KD, Domire ZJ, DeVita P, Rider PM, Kulas AS. The effect of
Nordic hamstring strength training on muscle architecture, stiff-
ness, and strength. Eur J Appl Physiol 2017;117:943–953.
Simpson CL, Kim BDH, Bourcet MR, Jones GR, Jakobi JM. Stretch
training induces unequal adaptation in muscle fascicles and thick-
ness in medial and lateral gastrocnemii. Scand J Med Sci Sports
2017;27:1597–1604.
Sinha S, Sinha U. Reproducibility analysis of diffusion tensor indices
and fiber architecture of human calf muscles in vivo at 1.5 Tesla in
neutral and plantarflexed ankle positions at rest. J Magn Reson
Imaging 2011;34:107–119.
Sinha U, Sinha S, Hodgson JA, Edgerton RV. Human soleus muscle
architecture at different ankle joint angles from magnetic resonance
diffusion tensor imaging. J Appl Physiol 2011;110:807–819.
Sinha U, Csapo R, Malis V, Xue Y, Sinha S. Age-related differences in
diffusion tensor indices and fiber architecture in the medial and lat-
eral gastrocnemius. J Magn Reson Imaging 2015;41:941–953.
Taylor DG, Bushell MC. The spatial mapping of translational diffusion
coefficients by the NMR imaging technique. Phys Med Biol
1985;30:345–349.
Ticinesi A, Meschi T, Narici MV, Lauretani F, Maggio M. Muscle
ultrasound and sarcopenia in older individuals: A clinical perspec-
tive. J Am Med Dir Assoc 2017;18:290–300.
Treece GM, Prager RW, Gee AH, Berman L. Fast surface and volume
estimation from non-parallel cross-sections, for freehand three-
dimensional ultrasound. Med Image Anal 1999;3:141–173.
Treece GM, Prager RW, Gee AH, Berman L. Surface interpolation
from sparse cross sections using region correspondence. IEEE
Trans Med Imaging 2000;19:1106–1114.
Treece GM, Gee AH, Prager RW, Cash CJ, Berman LH. High-
definition freehand 3-D ultrasound. Ultrasound Med Biol
2003;29:529–546.
Vernillo G, Pisoni C, Sconfienza LM, Thiebat G, Longo S.
Changes in muscle architecture of vastus lateralis muscle
after an Alpine snowboarding race. J Strength Cond Res
2017;31:254–259.
Wakeling JM, Jackman M, Namburete AI. The effect of external com-
pression on the mechanics of muscle contraction. J Appl Biomech
2013;29:360–364.
2504 Ultrasound in Medicine & Biology
Weng L, Tirumalai AP, Lowery CM, Nock LF, Gustafson DE, Von
Behren PL, Kim JH. US extended-field-of-view imaging technol-
ogy. Radiology 1997;203:877–880.
Wickiewicz TL, Roy RR, Powell PL, Edgerton VR. Muscle architec-
ture of the human lower limb. Clin Orthop Relat Res
1983;179:275–283.
Yanagisawa O, Kurihara T, Okumura K, Fukubayashi T. Effects of
strenuous exercise with eccentric muscle contraction: Physiological
Volume 44, Number 12, 2018
and functional aspects of human skeletal muscle. Magn Reson Med
Sci 2010;9:179–186.
Zaraiskaya T, Kumbhare D, Noseworthy MD. Diffusion tensor imaging
in evaluation of human skeletal muscle injury. J Magn Reson Imag-
ing 2006;24:402–408.
Zhang J, Zhang G, Morrison B, Mori S, Sheikh KA. Magnetic reso-
nance imaging of mouse skeletal muscle to measure denervation
atrophy. Exp Neurol 2008;212:448–457.