C H A P T E R
10
Imaging
Roger E. Price
Small Animal Cancer Imaging Research Facility,
U.T.MD Anderson Cancer Center, Houston,TX, USA
Introduction
Advances in imaging technology now allow high resolution
noninvasive and nondestructive imaging of mice with
body weights of less than 30 g by magnetic resonance
(MR) at 50 ␮m resolution in vivo and 10 ␮m in vitro,
by computed tomography (CT) at 25 ␮m in plane res-
olution, and positron emission tomography (PET) at
1–2 mm resolutions (Johnson et al., 1993; Budinger
et al., 1999; Paulus et al., 2000, 2001; Balaban and
Hampshire 2001). Similar technological advances are also
being made in optical, ultrasound, and radiographic
imaging. Imaging modalities also uniquely allow func-
tional information to be obtained and viewed in the con-
text of anatomic information. Judicious exploitation of
the unique benefits of each imaging modality and the
combination of information from multiple imaging
modalities will permit expeditious economical pheno-
typing of animal models for mutations, pathologic
changes, disease progression, and response to therapy.
Monitoring tissue characteristics using non-
destructive imaging methods makes it feasible to con-
duct longitudinal studies on laboratory animals. The
ability to acquire multiple data points and monitor dis-
ease evolution within an individual animal can reduce
The Laboratory Mouse
Copyright 2004 Elsevier
ISBN 0-1233-6425-6
IMAGING
the number of animals required to compensate for bio-
logical and inter-individual variability in order to
obtain significantly relevant data in experimental ani- 167
mal investigations. Noninvasive imaging methods can
also be used for accurate determination of optimal
ANATOMY AND DEVELOPMENTAL BIOLOGY
times for interventional treatments or for tissue harvest
and pathology, thus further reducing the number of
animals required in animal research. In addition to the
moral implications of reducing the number of animals
required in research studies, the financial saving result-
ing from the reduced animal purchase and mainte-
nance costs and the reduced space and personnel
required to house and care for research animals, further
mitigates the cost of performing the imaging studies.
Magnetic resonance imaging is a versatile, fast,
non-destructive imaging technique that is capable of
providing high resolution images with excellent soft
tissue contrast and detailed information on tissue char-
acteristics, anatomy, and function. The inherent
isotropic three-dimensional nature of MR imaging
allows detailed analysis of individual organs and retro-
spective studies through any arbitrary plane ( Johnson
et al., 1993). Recent availability of increasing magnetic
field and gradient strengths are producing dramatic
improvements in signal-to-noise ratios and spatial and
temporal resolution (Allport and Weissleder 2001).
All rights of production in any form reserved
 The images in this chapter were acquired using a
4.7 T, 40 cm bore Bruker Biospec Avance MR scanner
(Bruker Instruments, Karlsruhe, Germany) in the
Small Animal Cancer Imaging Research Facility at
The University of Texas M.D. Anderson Cancer
Center Houston, Texas, USA. All images were acquired
using a 6 cm actively shielded gradient system (950
mT/m, 110 ␮s rise time, and 8636 T/m/s slew rate)
and a 25 mm linear volume resonator. Three regions of
the animals were imaged (head, thorax, and abdomen).
Axial, T1-, T2-, and proton density-weighted images
were acquired from each region. The axial head images
covered a 2.5 cm square field-of-view (FOV), while the
sagittal head and axial thoracic, abdominal, and pelvic
images had an FOV of 3 cm. Data was acquired at
512 ⫻ 256 data matrices then zero-filled to 512 ⫻ 512
before Fourier Transform, resulting in an isotropic in
plane resolution of 59 ␮m for all axial images except
those of the head, which were 49 ␮m resolution. All
slices were 1 mm thick and separated by 0.25 mm
gaps. The T1-weighted images (TE/TR ⫽ 10.3/905
IMAGING
ms) were acquired in 1 h, while the T2-weighted images
(TE/TR ⫽ 55/2200 ms) required 2.5 h. The proton
168
ANATOMY AND DEVELOPMENTAL BIOLOGY
Figure 10.1 Midline sagittal images of head.
1. Cervical Vertebra II (Axis) 6. Basisphenoidal Bone
2. Cervical Vertebra I (Atlas) 7. Basilar Portion of Occipital Bone
3. Cerebellum 8. Pituitary
4. Cerebrum 9. Trachea
5. Presphenoid Bone 10. Epiglottis
density weighted images (TE/TR ⫽ 10.3/5000 ms)
were acquired in 1.5 h. Magnetic resonance images
were displayed with the Image Analysis software
Analyze® (CN Software, Southwater, UK) on a Silicon
Graphics O2 computer for presentation.
Images of the head and thoracic regions were
acquired using adult female Hsd:ICR (CD-1®) mice
(Charles Rivers Laboratories, Inc., Wilmington,
Massachusetts, USA) and images of the abdomen and
pelvic region of male mice were acquired from adult
NOD.CB17-Prkdc scid/J mice (The Jackson Laboratory,
Bar Harbor, Maine, USA). T2-weighted images of the
head were acquired on an unconscious animal which
was immobilized using isoflurane inhalation anesthesia.
For the remainder of the images of the head and other
regions of the body, the animals were euthanized with
CO2 and imaged dead (to eliminate motion artifacts).
Following imaging, the mice were left in the sleds in
which they were imaged and quick frozen in a ⫺70⬚C
freezer until rigid. Once frozen the imaged regions were
cut in to transverse blocks of tissue for the axial images
and into a sagittal block for the sagittal images of the
head on a band saw and embedded in OCT embedding
11. Basihyoideum
12. Salivary Glands
13. Nasal Cavity/Nasal Pharynx
Figure 10.2 Midline sagittal T1-weighted MR images with graphic slice locations for each imaged region.

IMAGING
169

Figure 10.3 Axial images of head (Slice location A).

ANATOMY AND DEVELOPMENTAL BIOLOGY

1. Eye 4. Nasopharynx 7. Oral Cavity
2. Lacrimal Gland 5. Mandible 8. Endoturbinate
3. Maxilla 6. Tongue 9. Olfactory Bulb of Brain

Figure 10.4 Axial images of head (Slice location B).

1. Temporomandibular Joint 5. Caudal Cerebral Artery 9. Vertical Ramus of Mandible
2. Dorsal Sagittal Sinus 6. Maxillary Vein 10. Linguofacial Vein
3. External Capsule 7. Trigeminal Nerve
4. Lateral Ventricle 8. Nasopharynx
 Figure 10.5 Axial images of head (Slice location C).
1. Dorsal Sagittal Sinus 4. Basisphenoidal Bone 8. Mandibular Lymph Node
2. Corpus Collosum and Internal 5. Trigeminal Nerve 9. Mandibular Salivary Gland
Capsule 6. Nasopharynx
3. Temporal Vein 7. Mandible
IMAGING

170
ANATOMY AND DEVELOPMENTAL BIOLOGY

Figure 10.6 Axial images of head (Slice location D).

1. Inner Ear Portion of Petros Temporal 3. Superficial Temporal Vein 6. Sublingual Salivary Gland
Bone 4. Tympanic Membrane 7. Submandibular Salivary Gland
2. External Ear Canal 5. Maxillary Vein

Figure 10.7 Axial images of thorax (Slice location E).

1. Brown Fat 5. Rib 9. Sternebra
2. Scapula 6. Lung 10. Thoracic Lobe of Thymus
3. Thoracic Vertebra 7. External Jugular Vein
4. Thoracic Spinal Cord 8. Cervical Mammary Gland
Figure 10.8 Axial images of thorax (Slice location F).
1. Brown Fat 5. Lung 9. Heart, Interventricular Septum
2. Thoracic Spinal Cord 6. Thoracic Mammary Gland 10. Heart, Papillary Muscle Left Ventricle
3. Rib 7. Thymus 11. Sternebra
4. Trachea 8. Heart, Left Ventricle 12. Heart, Right Ventricle

IMAGING
171

ANATOMY AND DEVELOPMENTAL BIOLOGY

Figure 10.9 Axial images of thorax (Slice location G).
1. Mammary Gland 5. Vertebra 9. Posterior Vena Cava
2. Epaxial Muscle 6. Lung 10. Hepatic Vein
3. Spinal Cord (Gray Matter) 7. Rib 11. Gall Bladder
4. Spinal Cord (White Matter) 8. Esophagus 12. Liver

Figure 10.10 Axial images of abdomen (Slice location H).

1. Right Crus of Diaphragm 6. Right Adrenal Gland 11. Portal Vein
2. Spleen 7. Right Kidney 12. Stomach, Pylorus
3. Stomach, Fundus 8. Aorta 13. Duodenum
4. Spinal Cord 9. Caudal Vena Cava 14. Liver
5. Right Psoas Major Muscle 10. Pancreas 15. Jejunum
 Figure 10.11 Axial images of abdomen (Slice location I).
1. Stomach, Fundus 5. Caudal Vena Cava 9. Liver
2. Left Kidney 6. Right Renal Vein 10. Colon
3. Spinal Cord 7. Right Kidney, Cortex 11. Portal Vein
4. Left Psoas Major 8. Right Kidney, Papilla 12. Jejunum
IMAGING

172

Figure 10.12 Axial images of abdomen (Slice location J). Slight differences in the positions and sizes of organs between
ANATOMY AND DEVELOPMENTAL BIOLOGY

tissue block face image and MR images due to movement of caudal abdominal organs during freezing process.
1. Vesicular Gland 4. Left Ilium, Body 8. Right Deferent Duct
2. Descending Colon 5. Transverse Process, Lumbar Vertebra 9. Urinary Bladder
3. Superficial Circumflex Iliac Artery 6. Spinal Cord 10. Abdominal Fat
and Vein 7. Coagulation Gland

Figure 10.13 Axial images of abdomen (Slice location K).

1. Left Deferent Duct 5. Coagulation Gland 10. Right Ilium, Body
2. Left External Oblique Muscle of 6. Vesicular Gland 11. Right Subiliac Lymph Node
Abdomen 7. Descending Colon 12. Urinary Bladder
3. Left Subiliac Lymph Node 8. Left Ilium, Body
4. Duct of Vesicular Gland 9. Spinal Cord
medium for frozen tissue specimens (Tissue-Tek,
Sakura, Torrance, CA). The embedded blocks of tissue
were cryosectioned at ⫺20⬚C through the axial plane
of all three regions of the animal and the sagittal plane
of the head in 250–500 ␮m increments. At each level,
a high-resolution block-face digital image of the surface
of the frozen block of tissue was captured using an
Olympus 4-megapixel Camedia E-10 digital camera
with a macro lens. In all images, Tissue ⫽ block-face
image of frozen tissue block, PW ⫽ proton-weighted MR
image, T1 ⫽ T1-weighted MR image, T2 ⫽ T2-weighted
MR image (Figures 10.1–10.13).
Acknowledgments
The author would like to thank Belinda Rivera, B.S.,
R.V.T. for assistance in animal support and tissue pro-
cessing, Jim Bankson, Ph.D. for acquiring the MR
images, and Ms Sara Amra for cutting the frozen blocks
of tissue for the tissue images. The work described herein
was supported by The University of Texas M.D.
Anderson Cancer Center Support Grant (CA16672)
from the National Institutes of Health, National Cancer
Institute, Department of Health and Human Services.
References
Allport, J.R. and Weissleder, R. (2001). Exp. Hematol. 29,
1237–1246.
Balaban, R.S. and Hampshire, V.A. (2001). ILAR J. 42,
248–262.
Budinger, T.F., Benaron, D.A. and Koretsky, A.P. (1999).
Annu. Rev. Biomed. Eng. 1, 611–648.
Johnson, G.A., Benveniste, H., Black, R.D., Hedlund, L.W.,
Maronpot, R.R. and Smith, B.R. (1993). Magn. Reson. Q.
9, 1–30.
Paulus, M.J., Gleason, S.S., Kennel, S.J., Hunsicker, P.R. and
Johnson, D.K. (2000). Neoplasia 2, 62–70.
Paulus, M.J., Gleason, S.S., Easterly, M.E. and Foltz, C.J.
(2001). Lab. Anim. (NY) 30, 36–45.
Popesko, P., Rajtova, V. and Horak, J. (1992). A Color Atlas
IMAGING
of the Anatomy of Small Laboratory Animals 2. Mouse.
Wolfe Publishing Ltd., London.
Takamasa, I., Hiroshi, Y. and Toshiyuki, H. (2001). A Color
Atlas of the Sectional Anatomy of the Mouse. Adthree, Inc.,
Japan.
173
ANATOMY AND DEVELOPMENTAL BIOLOGY