ARTICLE IN PRESS

Ultrasound in Med. & Biol., Vol. 00, No. 00, pp. 112, 2018
Copyright © 2018 World Federation for Ultrasound in Medicine & Biology. All rights reserved.
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https://doi.org/10.1016/j.ultrasmedbio.2018.11.001

Original Contribution

EVALUATION OF BRAIN TUMOR IN SMALL ANIMALS USING PLANE WAVE-BASED

POWER DOPPLER IMAGING

TAGEDPJINGJING XIA,* YI YANG,* CHENWENBAO HU,* RUI MENG,* QIUJU JIANG,*,y RONG LIU,*,y YANYAN YU,*,y
ZONGHAI SHENG,*,y FEI YAN,* LIJUAN ZHANG,* ZHIFENG SHI,z HAIRONG ZHENG,*,y and WEIBAO QIU*,yTAGEDEN
* Paul C. Lauterbur Research Center for Biomedical Imaging, Shenzhen Institutes of Advanced Technology, Chinese Academy of
Sciences, Shenzhen, China; y Shenzhen Key laboratory of Ultrasound Imaging and Therapy, Shenzhen, China; and z Shanghai
Pituitary Tumor Center, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China
(Received 15 January 2018; revised 27 September 2018; in final from 3 November 2018)

Abstract—Precisely evaluating the characteristics of a glioma tumor in vivo is challenging when performing sur-
gical resection clinically. The infiltration characteristics of a tumor make precise resection difficult because of
uncertainties about the surrounding vasculature and the relationships with functional structures. Magnetic reso-
nance imaging is routinely used to distinguish the area of a glioma, but it cannot resolve details of the vascular
network around or inside the tumor. Ultrasound imaging is a real-time imaging modality that has been applied
clinically in intra-operative surgery, and the sensitivity of flow measurements in the brain is improved by ultra-
fast plane wave imaging. This study applies a plane wave-based power Doppler imaging method to visualize the
blood flow distribution in glioma models in vivo. This new imaging method makes it possible to delineate the flow
structure of a glioma tumor in the brain of a small animal. The tumor can be distinguished from normal brain tis-
sue, and different sections of the tumor contain different flow structures. The normalized blood flow intensities
(mean § standard deviation) within regions of interest were 0.33 § 0.13, 0.72 § 0.15, 0.36 § 0.23 and 0.06 § 0.07
for the type I normal rat, type I glioma rat, type II normal rat and type II glioma rat, respectively. Quantification
analysis verified the feasibility of using this plane wave-based Doppler imaging method to evaluate brain tumors
in small animals. (E-mail: [email protected]) © 2018 World Federation for Ultrasound in Medicine & Biology.
All rights reserved.
Key Words: Glioma, Plane wave ultrasound, Power Doppler imaging, Flow measurement, In vivo imaging.

INTRODUCTION resection with avoiding new deficits or neurologic

sequelae. Moreover, relapse is possible if the tumor tis-
Gliomas constitute about 80% of all malignant brain
sue is not removed thoroughly (Chamberlain 2011).
tumors (Goodenberger and Jenkins 2012). These are
Therefore, imaging guidance providing information
highly vascular tumors that cause major morbidity and
about vessel orientations and tumor boundaries is crucial
mortality because of their extreme aggressiveness and
during the surgery to avoid both major brain damage and
strong infiltration (Huse and Holland 2010). Although
recurrence.
surgical resection is considered the optimal treatment,
Magnetic resonance imaging (MRI) is a routine
surgery is particularly challenging because of the com-
method used to provide an overview of the intracranial
plex anatomy of such tumors, including the surrounding
contents and details of a glioma tumor by pinpointing
vasculature and the relationship to functional structures
the structures and their connections to nearby tissues
(Sanai et al. 2010). The tumor may be located within or
(Zhou et al. 2011). It has been applied both pre-opera-
adjacent to neural areas providing critical language and
tively and intra-operatively to determine the macro-
motor functions (Sanai and Berger 2010; Sanai et al.
scopic outline of a glioma to aid surgical resection
2008), which makes it difficult to balance extensive
(Hatiboglu et al. 2009; Hygino da Cruz et al. 2011).
Address correspondence to: Weibao Qiu, Paul C. Lauterbur
However, although MRI allows the area of a glioma to
Research Center for Biomedical Imaging, Shenzhen Institutes of be distinguished, it cannot be used to resolve the
Advanced Technology, Chinese Academy of Sciences, 1068 Xueyuan details of the infiltration and vascular network inside
Avenue, Nanshan, Shenzhen 518055, China. E-mail:
[email protected]
or around the tumor. The image resolution of a 3-T

1
 ARTICLE IN PRESS
2 Ultrasound in Medicine & Biology Volume 00, Number 00, 2018

MRI is usually about 1 mm (Geerligs et al. 2017), METHODS

which is not high enough to evaluate the tumor in a
Imaging strategies
small animal. In addition, MR-based cerebral blood
Figure 1 illustrates our experimental setup and data
volume (CBV) mapping has good spatial resolution but
acquisition scheme. The imaging data were acquired by
limited temporal resolution and raises practical issues
a Verasonics Vantage system (Verasonics, Redmond,
in the laboratory environment, and intra-operative MRI
WA, USA) comprising a full-channel electronic module
is expensive and time consuming (Gerganov et al.
for plane wave ultrasound transmission and reception. A
2011).
linear-array transducer with a center frequency of
Ultrasound is a safe, real-time and economic imag-
15 MHz (128 elements, 0.1-mm pitch; L15-128, Ver-
ing modality that has been applied intensively in clinical
mon, France) was connected to the data acquisition sys-
practice (Chen et al. 2016). Traditional gray-scale imag-
tem via a customized adaptor board. To obtain images of
ing using intra-operative ultrasound is limited mainly to
the cerebral blood flow, the probe was fixed on a stage
localization and characterization of pathologic features,
and real-time B-mode imaging was used to guide the
providing information for use when determining the
imaging of the appropriate imaging sections. Raw radio-
extent of tumor resection (Renner et al. 2005; Selbekk
frequency (RF) data were saved on the hard disk of a
et al. 2013). Although traditional ultrasound can provide
computer for further image processing.
little information on vascularization in the brain, con-
For the scan scheme, we developed a plane wave
trast-enhanced (CEUS) ultrasound has been used to
imaging sequence of 11 tilt angles for coherence com-
detect tumor angiogenesis in the brain (Ellegala et al.
pounding covering a total angular range of 21˚ to scan
2003; Orringer et al. 2010). This is a complementary
the entire brain. We transmitted three pulses at each
method for recognizing blood vessels before or during
plane wave transmission and received the echoes three
tumor resection, but the perfusion time of the contrast
times for averaging to reduce background noise. The
agent needs to be allowed for.
averaged signal was considered as the final echo for
The recently developed method of plane wave-
the delay-and-sum beamforming process to form an
based power Doppler ultrasound flow imaging has been
image at one tilt angle. The combined frame rate was
used for visualizing brain vessels by transmitting and
therefore
1000 frames/s.
receiving parallel data with all the array elements syn-
chronously rather than conventional line-by-line scan-
ning (Mac
e et al. 2011, 2013; Urban et al. 2015). Very
Preparation of the animal model
high frame rate ultrasound imaging data (>10,000
All animal procedures were approved by the Shenz-
frames/s) (Tanter and Fink 2014) were acquired to
hen Institutes of Advanced Technology, Chinese Acad-
enable high-sensitivity power Doppler imaging process-
emy of Science (Approval No. SIAT-IRB-150529-YGS-
ing (Montaldo et al. 2010). The sensitivity of flow mea-
QWB-A0088-02). In vivo studies were conducted on
surement was significantly improved by compounding
female 7-wk-old Sprague-Dawley rats that weighed
multiple imaging planes with titled angles. This elevated
approximately 200 g. The C6 glioma model was used as
sensitivity allowed functional ultrasound to track the
spatiotemporal dynamics of the activation within the rat Linear transducer
brain after whisker stimulation or during induced epilep-
Skull window
tic seizures (Mac
e et al. 2011). It was also reported to be
Sprague-Dawley rat fixed on
sensitive enough to detect odor-evoked stimulation 3D stereotaxic apparatus
(Osmanski et al. 2012) and, remarkably, to map func-
tional connectivity in the living rat brain with a spatio- Y
Coronal plane of the brain
temporal resolution much higher than that of functional
Z X
MRI (fMRI) (Osmanski et al. 2014).
In the present study, we took advantage of the high
sensitivity of plane wave-based Doppler ultrasound flow
imaging to visualize the blood flow distribution of a gli-
oma tumor in a small animal for the first time, yielding a 300 frames compounded

highly resolved neurovascular map of the glioma. The 11 angles per frame

flow images were optimized with Tukey window and

dynamic receive aperture technology. In vivo small ani-
Cerebral blood flow map
mal experiments were performed to illustrate the perfor-
mance of the proposed new method in evaluating glioma Fig. 1. Framework of this study, illustrating the principle
tumors. underlying plane wave Doppler imaging.
 ARTICLE IN PRESS
Brain tumor evaluation in small animals
J. XIA et al. 3

an orthotopic brain tumor. The rats were injected with method, and can efficiently remove clutter signals and
C6 cells in the right brain 3 mm laterally and 1 mm preserve blood flow signals even at low frequencies by
axially from the bregma point (Valable et al. 2007). A removing the proper eigen-components (Osmanski et al.
burr hole with a diameter of 1 mm was drilled, and 10 2014).
mL of cell suspension (106 cells/mL) was injected into We define the 300-IQ data packet as a 3-D matrix S
the brain within 10 min using a micro-injection driven (x, z, t), where (x, z) is the physical coordinates of the
by a Hamilton syringe (KD Scientific, Holliston, MA, pixel, and t is time. The mathematical expression of the
USA). The needle was slowly removed 5 min after the SVD filter (extracting the blood signal from S(x, z, t), con-
injection, and then the burr hole was sealed with bone verting the 3-D blood signal Sblood(x, z, t) to a 2-D matrix
wax and the scalp was sutured. MR images were Sblood(k, t) and applying the SVD decomposition) is
obtained before the experiment to ensure successful
X
N
tumor formation. If the tumor had been generated suc- Sblood ðx; z; t Þ ¼ ei  Bi ðx; zÞ  Si ðt Þ ð1Þ
cessfully, ultrasound imaging was performed 0.51 d i ¼ 1
after the MRI scan. There were 50 rats in total, and 28 of
them were selected for the study (16 rats failed without The matrix Bi(x, z) will give the spatial distribution
tumor, and 6 rats died during the skull surgery). of the signal Si(t) for the corresponding singular value of
For the ultrasound imaging experiment, the rat was ei ði ¼ 1; 2; . . . ; NÞ. The asterisk denotes the multipli-
shaved and then fixed on a 3-D stereotaxic apparatus cation. Based on the previous research, the tissue move-
(68028, RWD, Shenzhen, China) after anesthetization ments generate coherent signals while the vascular
with 10% chloral hydrate. However, although ultrasound signals coming from the randomly moving red blood cells
can propagate deep within tissues, cerebral US remains are uncorrelated. The larger singular values correspond to
limited by its poor penetration through the skull. An the tissue signal, and the intermediate singular values cor-
attenuation of 6.9 dB/cm at 15 MHz has been observed respond to the blood signal; the last few singular values
in previous studies (Fry and Barger 1978; Larrat et al. represent the noise signal (Song et al. 2016). Then, we
2010; Pinton et al. 2012), which renders small variations used the low-order and high-order singular value thresh-
in blood flow indistinguishable from noise. Therefore, to olds (90 and 270, respectively) to remove the markedly
avoid ultrasound attenuation by the skull, the skull was low and high singular values and extract the blood signal.
drilled over an area of approximately 0.8 £ 1.2 cm Additionally, a Butterworth high-pass filter with a
(bilateral window), with the dura remaining intact, and 25-Hz cutoff frequency was employed to reduce the
was sprayed with saline water for cleaning out and cool- noise further. Finally, we calculated the square of the fil-
ing during the drilling process. The whole brain could tered IQ data and averaged over the total 300 matrix to
then be imaged through the drilled window by an ultra- form one blood flow map. We thus acquired information
sound transducer with the aid of a coupling agent. on cerebral coronal blood flow, including the vascular
structure of the glioma tumor.
Ultrasound imaging processing To improve image quality, we also proposed the
All data were post-processed offline using MAT- dynamic receive aperture imaging technology for the
LAB (MathWorks, Natick, MA, USA). Delay-and-sum receive beamformer. We found that the sound field near
beamforming and coherence compounding yielded a the aperture fluctuates strongly, resulting in a interfer-
frame ensemble in the time domain, with 300 com- ence effect. Fortunately, these effects can be avoided by
pounded frames (the total acquisition time was 0.3 s) using smaller apertures in the near field (Harris 1978).
used as an ensemble to form a cerebral blood flow Compared with the beamformer using the boxcar win-
image, as illustrated in Figure 1. In detail, we first dows function, the use of dynamic aperture can maintain
obtained the beamformed RF data at each transmission a certain ratio between the effective aperture size and
angle with delay-and-sum beamforming and coherently the focusing depth. The equation, based on a boxcar win-
summed the RF data of all 11 titled angles to form one dow function, is
frame matrix. Second, we took 300 time-continuous 8  

>
compounded frames as an ensemble and applied Hilbert < 1; n  pitchx=z
fnumber =2
>
WðnÞ ¼   n ¼ 0 » N1
transfer to each frame matrix to obtain 300-continuous- >
>  
IQ data matrix. In the third step, a high-pass spatiotem- : 0; n  pitchx=z > fnumber =2
poral filter based on singular value decomposition
ð2Þ
(SVD) was applied to the 300-IQ data packet in the time
domain to adaptively suppress slow-time clutters of
low-frequency tissue signals. This adaptive SVD filter is where pitch means the distance between two adjacent
a robust and reliable orthogonal matrix decomposition elements, and (x, z) are the physical coordinates of the
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4 Ultrasound in Medicine & Biology Volume 00, Number 00, 2018

pixel. So n  pitchx represents the size of the effective RESULTS

aperture. The fixed value of fnumber is set to 1.75 in this
Image quality analysis
study. Window functions can suppress the amplitude of
By using a set of tilt-angle plane wave illuminations
grating lobes and reduce their energy (Harris 1978). In
and coherently summing the acquired set of images,
fact, the boxcar window is not the optimal choice for
compound ultrasound images were produced with better
improving image quality. Optional window functions,
resolution and lower noise compared with conventional
including the boxcar window, Hanning window, Ham-
images, while preserving a kilohertz frame rate. To
ming window and Tukey window, were compared in
ensure the best visualization of the micro-blood flow sig-
this study. The imaging performance of different win-
nal, the display dynamic range of the images were
dow functions was quantitatively analyzed.
adjusted carefully to an optimal value. Figure 2 illus-
To a certain extent, this proposed method can
trates the effect of dynamic receive aperture technology
improve the signal-to-noise (SNR) and contrast-to-noise
on blood flow images from a coronal view of an in vivo
of the Doppler images.
normal rat brain. It is clear that Figure 2b provides a bet-
ter visualization of the Doppler image. Quantitative
MRI data acquisition SNR values have been calculated as ratios of mean blood
Because MRI is a non-invasive medical imaging signal (indicated by the ROIs within the red-lined box)
tool and has been frequently used for the diagnosis of and standard deviation of the background noise (indi-
glioma tumors, here we compare the proposed imaging cated by the ROIs inside the green-lined box). The SNR
method with a traditional MRI system. Twelve hours values of the two images are 26.2 and 41.3 dB, respec-
before the ultrasound imaging experiments, we first tively, with 15.1-dB enhancement.
scanned the rat to confirm the existence of gliomas and Figure 3 illustrates the effect that different window
delineated the outline structure in the rat brain using functions have on rat cerebral blood flow images in the
T2-weighted MRI (MAGNETOM Trio, Siemens, Ger- coronal view (Fig. 3a). Figure 3b compares the weighted
many). A standard 3-T clinical scanner equipped with a curves of different window functions. We then plotted in
custom-designed three-axis balanced-torque head gradi- Figure 3(c, d) the lateral normalized intensity profiles in
ent coil was used. In MRI, “T2” refers to the transverse the red-lined box in Figure 3a processed under different
relaxation time, and different tissues have different window functions of three selected vessels. The intensity
T2 values during an MRI scan, with a longer T2 result- profiles in Figure 3c were normalized separately, and the
ing in a higher signal intensity. The normal left rat brain intensities in Figure 3d were normalized uniformly. The
and normal right rat brain appear almost identical in red curve represents the profile in the absence of a win-
T2-weighted images, whereas these can differ in appear- dow function. It can be concluded that window functions
ance in a rat with glioma. The area of a glioma would can suppress the grating lobes. In the meantime, the
appear at a higher intensity than the other parts of the Tukey window (light blue curve) and boxcar window
brain, making it possible to determine the existence and (blue curve) provide better resolution at full width at
properties of the glioma tumor (Pirzkall et al. 2001). half maximum (normalized intensity = 0.5), as indicated
by the black arrows in Figure 3c. In contrast, the second
and third peaks of the Tukey window are higher than the
Histopathology
boxcar window as indicated by the black arrows in
Immediately after acquiring the cerebral blood flow
Figure 3d. Considering the resolution and signal inten-
information, we performed transcardial perfusion sur-
sity, the Tukey window can provide the best imaging
gery (Gage et al. 2012) and prepared the brain sections.
performance. Therefore, it has been used in the follow-
When we were slicing the brain along the coronal plane,
ing tests.
the target area was approximately 0.5 to 1.5 mm axi-
ally from the bregma point. The thickness of each sec-
tion was 30 mm. We used hematoxylin and eosin (HE) Flow imaging in a glioma tumor
to stain the coronal sections, and observed them at the The high sensitivity of this imaging method is uti-
cellular level under an electron microscope (Olympus, lized to acquire the blood flow distributions both inside
Japan). The glioma area appeared darker than the healthy and around the glioma tumor. Figure 4(a, c) are images
areas (Nakamura et al. 2004), because the glioma of a rat model that had been injected with C6 cells for
nucleus is larger and more likely to be hyperchromatic, 22 d. The B-mode images in Figure 4a (dynamic
and there are a large number of cells. These observations range = 50 dB) reveal a hyper-echoic area that is clearly
represent direct evidence of the existence of glioma. different from the contralateral part of the brain.
 ARTICLE IN PRESS
Brain tumor evaluation in small animals
J. XIA et al. 5

a b

1mm

Fig. 2. Effect of dynamic receive aperture technology on Doppler imaging. (a) Beamforming without dynamic aperture.
(b) Beamforming with dynamic aperture. The signal-to-noise value of (b) is 15.1 dB higher than that of (a).

According to the corresponding power Doppler image in accurate reference. Figure 5(a, b) are MRI images of two
Figure 4c (display dynamic range = 45 dB), the tumor glioma rats in the coronal view scanned 0.5 d before ultra-
part in the right brain has a greater flow and a higher sound imaging, and Figure 5 (c, d) are the corresponding
intensity compared with the contralateral area of the plane wave Doppler images of Figure 5(a, b) (display
brain. Moreover, the flow pattern in the tumor area fea- dynamic range = 45 dB), respectively. The
tures more curved segments than normal tissue. glioma tumors in the rats are not easy to locate in Figure 5
Figure 4(b, d) are images of another rat model that (a, b) because of the low resolution of MRI; it is even
had been injected with C6 cells for 26 d. Similarly to more difficult to obtain the flow information. However, in
Figure 4a, the B-mode image in Figure 4d (dynamic the ultrasound power Doppler images in Figure 5(c, d), it
range = 50 dB) illustrates hyper-echoic signals over a is possible to identify the tumor from the blood flow dis-
large area in the right part of the brain. The tumor area is tributions within the tumor region, in which blood flow
larger than for the brain injected for 22 d, which may structures differ from those in normal regions of the brain.
indicate a larger tumor. The corresponding power Dopp-
ler image in Figure 4d (display dynamic range = 45 dB) Different sections of the glioma tumor
differs from that in Figure 4c. The flow pattern is also Figure 6 illustrates nine coronal-view plane wave
quite different from that of the contralateral area of the Doppler images of a glioma tumor in different sections.
brain, and the number and intensity of hyper-echoic sig- We obtained blood flow images of about 95 continuous
nals for blood flow is clearly decreased in a large area, sections along the direction from bregma to lambda. The
which may indicate some changes in the tumor. distance between two adjacent sections is 100 mm. We
Although this region still exhibits hyper-echogenicity, then chose several representative sections: one for every
tumor necrosis might already be present because of an five from section 15 to section 55. As for the blood flow
insufficient blood supply. structure in the tumor area in the right brain, there is a
tendency in this series of section images for the blood
Comparison with MRI flow to appear rich on both sides of the tumor, with
Magnetic resonance scanning was used on glioma necrosis arising gradually in the middle parts. It could be
rats for the first time at about 15 d after injection of C6 inferred that sections 15 to 25 and sections 45 to 55 are
cells to confirm tumor growth. Generally, a 2- to 3-mm near the edges of the tumor. The blood flow intensity is
diameter of tumor in the dorsal view is optimal, and then greater because new blood vessels are needed to support
0.5 d before the ultrafast ultrasound imaging experiment, tumor growth. On the other hand, sections 30 to 40 are
the second MRI scan was conducted to provide a more in the central part of the tumor, and very low blood flow
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6 Ultrasound in Medicine & Biology Volume 00, Number 00, 2018

Fig. 3. Comparison between different window functions. (a) In vivo rat cerebral blood flow image in the coronal view.
(b) Weighted curves of the window functions in the central element. The lateral normalized intensity profile of the three
vessels in the red-lined box were processed under different window functions. (c) Intensity was normalized respectively.
(d) Intensity was normalized uniformly.

was detected because of the insufficient blood supply. structure. The two ROIs in the same images are totally
These observations indicate the feasibility of using the symmetric and the same size.
proposed method to perform vascular imaging of a gli- The results plotted in Figure 7a are normalized
oma tumor. We used V3-D software (Peng et al. 2010) intensities calculated for 13 type I samples and 15 type
to reconstruct the series of sectional images into a 3-D II samples. The normalized intensities were 0.33 §
image in Supplementary Video S1 (online only). 0.13 (mean § SD), 0.72 § 0.15, 0.36 § 0.23 and 0.06
§ 0.07 for the type I normal ROI, type I glioma ROI,
Flow quantification type II normal ROI and type II glioma ROI, respec-
It has been found that the tumor region appears as a tively.
hyper-echoic area in B-mode images and that its blood Before analyzing this statistical result, we also
flow structures can be distinguished from those in the used 25 samples of the normal rat model to calculate
normal regions of the brain, with these differences vary- and normalize the mean flow intensities of the ROIs in
ing with tumor growth (tumor volume). Specifically, the left and right brain. In Figure 7b it is seen that the
compared with normal cerebral blood flow, the tumor intensities in the two ROIs are 0.44 § 0.06 and 0.49 §
flow structures could be divided into two types: (type I) 0.04, respectively. This indicates that the mean intensi-
regions with greater blood flow and (type II) regions ties in the left and right brain of normal rats are nearly
with poor blood flow. We calculated the mean flow equivalent. On the basis of this result, it can be proved
intensities by manually choosing two ROIs (indicated by that the intensities in the tumor area of the type I rat
the red-dashed squares in Fig. 4) within the tumor and model are much higher than those in the normal area,
the normal cerebral region for both types of flow and on the other hand, the intensities in the tumor area
 ARTICLE IN PRESS
Brain tumor evaluation in small animals
J. XIA et al. 7

a b

c d

1 mm

Fig. 4. Comparison between plane wave B-mode images and plane wave ultrasound Doppler images of brain tissue with
a glioma tumor. (a) B-Mode image from plane wave ultrasonography of a 22-d-old tumor in the right brain. (b) B-Mode
image from plane wave ultrasonography of a 26-d-old tumor in the right brain. (c) Blood flow image of the brain in (a).
(d) Blood flow image of the brain in (b).

of the type II rat model are much lower than those in the tissue in the stained region, consistent with the data
normal area. acquired with other imaging modalities.

Histology comparison
DISCUSSION
Hematoxylin and eosin staining was performed to
enable precise verification of glioma tumor. Figure 8 Precisely evaluating the characteristics of a glioma
compares the (a) MR image, (b) histology image, (c) tumor in vivo is a challenge when performing surgical
B-mode image and (d) plane wave power Doppler image resection clinically. Knowledge of the surrounding vascu-
of a glioma rat. As illustrated in the MR, B-mode and lature of the tumor is necessary to avoid both major brain
flow images, the glioma tumor grows in the right brain damage and recurrence. Ultrasound Doppler imaging
of this rat. Figure 8b is an image of the HE-stained slice method is a well-established tool for flow analysis and
about +1.92 mm from bregma scanned by electron quantification. A recently developed plane wave flow
microscopy panoramic. As we can see, the glioma tumor imaging method exhibits improved sensitivity (Mac
e et
area in the right brain is much darker than the normal al. 2011, 2013). Plane wave-based contrast-enhanced
 ARTICLE IN PRESS
8 Ultrasound in Medicine & Biology Volume 00, Number 00, 2018

a b

c d

1 mm

Fig. 5. Comparison between traditional magnetic resonance images and plane wave ultrasound power Doppler images of
brain tissue with a glioma tumor. (a, b) Magnetic resonance images. (c, d) Blood-flow plane wave ultrasound images of
(a) and (b), respectively.

ultrasound CEUS presents several advantages over tradi- on a glioma tumor (Figs. 4 and 5), which makes it a valu-
tional ultrasound and can provide high-resolution images able tool for tumor characterization. A dynamic receive
of the vascular network of a rabbit kidney (Tremblay- aperture technology was developed to further improve
Darveau et al. 2014). Moreover, multi-frame-rate plane the imaging resolution of power Doppler measurement.
wave CEUS was developed for vasculature imaging in We quantitatively analyzed the effects of different win-
subcutaneous lymphoma and perfusion quantification dow functions on the receive beamformer in Figure 3,
(Leow et al. 2017). In addition, ultrasound localization and concluded that the Tukey window was an optimal
microscopy was utilized to visualize subcutaneous fibro- choice in suppressing the grating lobes and reducing
sarcoma angiogenesis (Lin et al. 2017). The present study their energy. The SNR value of the improved power
is the first to use plane wave ultrasound to obtain flow Doppler image was increased by 15.1 dB, as illustrated
information on a brain glioma tumor in a small animal in Figure 2.
in vivo. The image processing performed in this study
The proposed plane wave-based Doppler imaging revealed that glioma tumor regions exhibited blood flow
method can be used to obtain detailed flow information structures that differed from those in the normal regions
 ARTICLE IN PRESS
Brain tumor evaluation in small animals
J. XIA et al. 9

Section-15 Section-20 Section-25

Section-30 Section-35 Section-40

Section-45 Section-50 Section-55

1mm

Fig. 6. Plane wave-based Doppler images of the glioma at different imaging locations in coronal views: several repre-
sentative sections of 95 continuous section images, and the step between two adjacent sections is 100 mm.

of the brain. In addition, different tumor growth periods signal intensities of the blood flow in the normal and
may be associated with quite different blood flow distri- tumor regions in the same imaging sections and used sta-
butions in tumor regions. The experimental results indi- tistical analyses to identify different positions of the
cated that blood flow was poor in the central part of a tumor.
tumor that had been growing for a long time. The greater In this article, we described a newly proposed
intensity of blood flow in the tumor periphery may method for providing information on vessel orientations
reflect the aggressiveness of the tumor in forming new and distributions. It is an important reference for exten-
blood vessels to support its growth. Moreover, as the sive resection to avoid both major brain damage and
tumor grows, its central part may be affected by necrosis recurrence. Moreover, the imaging method proposed
if blood flow decreases significantly. We compared the here has the potential to be a useful tool in determining

a b
Normalized Intensity

Normalized Intensity

Fig. 7. Normalized intensity of blood flow in the tumor and normal cerebral blood flow. (a) In the rat model of glioma,
blood flow intensity is greater in type I than in type II glioma. (b) Comparison of blood flow in the left and right ROIs of
the normal rat model. ROI = region of interest.
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10 Ultrasound in Medicine & Biology Volume 00, Number 00, 2018

Fig. 8. Mapping the stained brain slice of a glioma rat to the corresponding B-mode image and plane wave Doppler
image. The tumor area is much darker and is consistent with the plane wave Doppler image. (a) Magnetic resonance
image. (b) Histology image. (c) B-Mode image (50 dB). (d) Plane wave power Doppler image.

whether patients with tumor are suited for targeted ther- flow image. A field-programmable gate array and GPU
apy, which would provide the basis for the follow-up for (graphics processing unit) could be used to expedite the
targeting therapy of tumor blood vessels and reducing process by implementing parallel computing (Qiu et al.
the suffering of patients. Multimodality imaging 2013, 2015). Clinical testing during intra-operative sur-
(Kircher et al. 2012) could be implemented by combin- gery in humans could be performed once the system
ing gray-scale B-mode imaging, MRI and the proposed becomes sufficiently fast. Moreover, the glioma tumor
plane wave flow imaging. Registration of local flow images obtained in different sections in coronal and sagit-
images of the tumor region with B-mode images is illus- tal views in Figure 6 indicated the feasibility of establish-
trated in Figure 9(a, c). The details of the vascular net- ing 3-D vascular imaging of a glioma tumor.
work inside the tumor could be resolved. Registration of
ultrasound flow images with MR images is illustrated in
CONCLUSIONS
Figure 9(b, d). Such a combination of imaging modali-
ties would allow full evaluation of the characteristics of This study is the first to characterize the blood flow
the tumor. Combined with other imaging guidance meth- distributions of a brain glioma tumor in a small animal
ods, a more accurate and safe surgery could be con- using plane wave Doppler imaging. Vascular maps of
ducted. the glioma were obtained, and detailed flow information
The ultrasound images acquired in this study were for the whole brain—notably inside and around the
obtained by processing RF data using an offline computer, tumor—could be acquired. This new method of plane
which was a time-consuming process to obtain a single wave ultrasound can be used to evaluate malignant
 ARTICLE IN PRESS
Brain tumor evaluation in small animals
J. XIA et al. 11

a b

c d

1 mm

Fig. 9. Registration of different imaging modalities. (a, b) Registration of ultrasound flow image with the B-mode image
(a) and the magnetic resonance image (b) for a 22-d tumor model. (c, d) Registration the ultrasound flow image with the
B-mode image (c) and the magnetic resonance image (d) for a 26-d tumor model.

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