Feasibility Study On Photoacoustic Guidance For High-Intensity Focused Ultrasound-Induced Hemostasis

Feasibility study on photoacoustic
guidance for high-intensity focused

ultrasound-induced hemostasis
Van Phuc Nguyen
Jeehyun Kim
Kang-lyeol Ha
Junghwan Oh
Hyun Wook Kang
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Journal of Biomedical Optics 19(10), 105010 (October 2014)
Feasibility study on photoacoustic guidance for

high-intensity focused ultrasound-induced
hemostasis
Van Phuc Nguyen,a Jeehyun Kim,b Kang-lyeol Ha,c Junghwan Oh,a,d and Hyun Wook Kanga,d,*
a
Pukyong National University, Interdisciplinary Program of Marine-Bio, Department of Electrical and Mechanical Engineering, 45 Yongso-ro,
Nam-Gu, Busan 608-737, Republic of Korea
b
Kyungpook National University, School of Electrical Engineering and Computer Science, 80 Daehakro, Bukgu, Daegu 702-701, Republic of Korea
c
Pukyong National University, Department of Physics, 45 Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea
d
Pukyong National University, Center for Marine-Integrated Biomedical Technology (BK 21 Plus), Department of Biomedical Engineering,
45 Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea
Abstract. The feasibility of photoacoustic imaging (PAI) application was evaluated to map punctured blood
vessels thermally treated by high-intensity focused ultrasound (HIFU) for hemostasis. A single-element
HIFU transducer with a central frequency of 2.0 MHz, was used to induce thermal hemostasis on the punctured
arteries. The HIFU-treated lesion was imaged and localized by high-contrast PAI guidance. The results showed
that complete hemostasis was achieved after treatment of the damaged blood vessels within 25 to 52 s at the
acoustic intensity of 3600 Wcm2 . The coagulation time for the animal artery was 20% longer than that of
the phantom possibly due to a lower Youngs modulus. The reconstructed PA images were able to distinguish
the treated area from the surrounding tissue in terms of augmented signal amplitudes (up to three times).
Spectroscopic studies demonstrated that the optimal imaging wavelength was found to be 700 nm in order
to reconstruct high-contrast photoacoustic images on HIFU-treated lesions. The proposed PAI integrated
with HIFU treatment can be a feasible application to obtain safe and rapid hemostasis for acute arterial bleeding.
2014 Society of Photo-Optical Instrumentation Engineers (SPIE) [DOI: 10.1117/1.JBO.19.10.105010]
Keywords: high-intensity focused ultrasound; photoacoustic imaging; hemostasis; coagulation; laser.

Paper 140350PRR received Jun. 4, 2014; revised manuscript received Sep. 15, 2014; accepted for publication Sep. 23, 2014; published online Oct. 29, 2014.
For a few decades, a variety of investigations on bleeding control (i.e., hemostasis) methods have clinically been performed
with clamping, suturing, and therapeutic devices such as laser
coagulation, plasma argon, etc.1,2 The primary purpose of
these devices is to achieve rapid hemostasis with less complication. However, these methods have still been performed in
an invasive manner (i.e., open surgery) and can hardly treat
the damaged internal organs that are deeply located in a body.
In particular, no clamping can be accessible to damaged tissue
components in microsizes such as the carotid artery, veins, and
nerves.3 Additionally, the current treatment approaches can
merely entail superficial treatments on damaged areas with
minimal coagulation in an axial direction.
Recently, high-intensity focused ultrasound (HIFU) has been
investigated as a noninvasive or minimally invasive thermal
therapeutic method to control acute hemorrhage and to treat
tumors deeply located inside the body.47 Ultrasound-induced
tissue coagulation is a technique that can obtain hemostasis in
an almost noninvasive manner without excessive heating and
adverse ion radiation effect on adjacent tissue, which were
often observed in laser or argon plasma applications. In previous
studies on acute hemorrhage control, HIFU was used to induce
the rapid temperature increase within the focused acoustic field,
leading to irreversible tissue coagulation and cell necrosis in
the treated region and achieving hemostasis for the targeted
tissues.8,9 Due to relatively lower acoustic intensities, the peripheral area around the targeted lesions could still remain less
damaged or undamaged. HIFU technology has also been
applied to a large number of benign and malign solid tumors
such as prostate cancer, liver, kidney, bone, and brain as well
as dissolution for ischemic stroke.1012 Specifically, HIFU technique could control hemostasis for severely damaged blood vessels, eventually reducing the risk of hemorrhage and treatment.13
In order to improve the efficacy of HIFU treatment and to
precisely deliver acoustic energy to the targeted lesion, the treatment procedure still needs to be monitored and evaluated in real
time. Lately, HIFU treatment has been carried out with imaging
guidance modalities such as magnetic resonance imaging (MRI)
and ultrasound imaging.14,15 However, MRI scanners are still
expensive and bulky systems with lengthy scanning time.
Ultrasound imaging systems are relatively cost-effective and
able to readily locate the injured tissue as well as to obtain faster
hemostasis than visual inspection (i.e., 25 s for Doppler ultrasound guidance versus 125 s for visual inspection).5,16 However,
ultrasound imaging still suffers from the lack of accuracy and
specificity as well as low image contrast particularly for
noninvasive localization and treatment evaluation.17 Thus, to
overcome the limitations of the current imaging systems, a
photoacoustic imaging (PAI) approach can be used as an
alternative imaging guidance modality to monitor and evaluate
the process of thermal therapeutics. PAI is a nonionizing,
*Address all correspondence to: Hyun Wook Kang, E-mail: wkang@pknu.ac.kr
0091-3286/2014/$25.00 2014 SPIE
Introduction
Journal of Biomedical Optics
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Nguyen et al.: Feasibility study on photoacoustic guidance for high-intensity focused ultrasound-induced hemostasis
noninvasive, cost-effective, and hybrid imaging technique that

can provide high optical absorption with high ultrasound resolution.18,19 Additionally, PAI imaging is capable of generating
structural information of hemoglobin or melanin as well as functional information such as total hemoglobin concentration or
blood flow with strong contrast and high spatial resolution.20,21
Compared to conventional imaging modalities, the primary advantage of PAI is to image the targeted tissue in more than 5 cm
depth along with spatial resolutions from 2 to 500 m22,23 and to
monitor the temperature profile in the thermal lesion generated
by HIFU.24 Therefore, the integration of high-contrast PAI with
HIFU therapy can be a feasible image-guided tool to facilitate
acute hemostasis in clinical applications.
In this study, the feasibility of HIFU application was investigated in terms of treating acute hemorrhage in tissue and
incorporating high-contrast PAI guidance in order to accurately
localize the treated lesions after HIFU application and thus to
minimize undesirable thermal injury. Acoustic fields of the
HIFU transducer in water and its thermal effects on tissue
were initially characterized with a hydrophone and thermocouple. The temporal development of the coagulation process was
measured and evaluated as a function of acoustic intensity in
phantom and animal tissues. After HIFU treatments, histological
analysis was performed and compared with photoacoustic (PA)
images on the targeted tissue.
2
2.1
Materials and Methods

Tissue Samples
Two groups of samples were used for HIFU-induced hemostasis

testing: phantom (N 25) and animal femoral arteries (N 5).
The phantom arteries used plastic tubes of an outer diameter of
2 mm, and the animal ones (i.e., outer diameter of up to 2 mm)
along with heparinized blood samples were procured from
chickens at a local slaughter house. Prior to the experiments,
an incision hole of 0.5 mm in diameter was punctured with
a needle on each artery sample to induce bleeding during
thermal hemostasis testing. A syringe pump (55-5920, Harward
Apparatus, Holliston, Massachusetts) was used to perfuse
blood through each sample at a constant flow rate of 20 mlh

in order to mimic physiological conditions. At the onset of
bleeding through the hole, high ultrasonic energy was immediately applied to the bleeder until the hole was covered with
coagulated blood (i.e., coagulum) due to thermal denaturation.
The complete hemostasis was confirmed in light of no leakage
throughout the artery sample as well as visual observation. In
turn, the time for completion of hemostasis, termed as coagulation time in this study, was evaluated under various conditions.
During HIFU application, each sample was immerged in a water
bath at 38C similar to in vivo environments. Degassed water
was used to prevent any cavitation in the bath and to provide
the impedance matching between HIFU transducer and target
tissue. In the case of animal tissue testing, each post-HIFU
treated tissue was imaged with a digital microscope (1.3 M
Dino-Lite pro, Anmo Electronics, New Taipei City, Taiwan)
to evaluate any collateral damage to the surrounding tissue.
From the captured images, the extent of the thermally denatured
lesion was circumferentially measured (N 5) with imaging
software (Image J, National Institutes of Health, Bethesda,
Maryland). In addition, histological analysis was post-experimentally performed to confirm hemostatic responses of the targeted tissue. The tested blood vessel samples were fixed in 10%
formalin prior to standard hematoxylin & eosin (H&E) staining.
Each sample was cut into a thickness of 4 m at the center of the
HIFU-treated area, and high resolution (100) digital images of
the cross-sectioned tissues were captured with a Leica microscope system (Leica DM 500, New York Microscope Company,
Hicksville, New York).
2.2
HIFU Application
Figure 1 depicts a schematic diagram of experimental setups for

HIFU-induced hemostasis. A piezoelectric HIFU transducer
(H-148, Sonic Concept, Woodinville, Washington) with a focal
length of 51.7 mm yielded ultrasound waves at a central frequency of 2.0 MHz. Continuous ultrasound waves were generated with a function generator (DDF 3010, Shanghai MCP,
Shanghai, China) and then amplified by an RF amplifier
(525LA, ENI, Rochester, New York). A blood-perfused sample
Fig. 1 Experimental setup of high-intensity focused ultrasound (HIFU)-induced thermal coagulation on

artery with assistance of photoacoustic imaging (PAI).
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was positioned underneath the transducer and then exposed to

focused acoustic fields. The HIFU transducer was immerged in a
water tank with the beam axis directed toward an incision hole to
yield acoustic fields within a sample. In order to achieve the
precise position of a focal spot, a three-dimensional plastic conical cap was designed, fabricated, and mounted on the HIFU
transducer. In turn, the focus of the transducer was simply
aligned with the location of the targeted hole. For phantom
experiments, the acoustic intensities at the focus ranged from
1100 to 4600 Wcm2 to identify the appropriate dosage for
HIFU treatment on animal tissue. Coagulation time, defined
as the time required for complete hemostasis, was measured
and compared as a function of acoustic intensity (N 5 per
condition). Based upon the phantom results, the acoustic intensity of 3600 Wcm2 was then selected and used for animal
artery experiments due to potentially rapid coagulation along
with minimal thermal injury. Additionally, the peak temperature
variations at the focus were monitored by a thermocouple
(GT307, Gilwoo Company, Seoul, Republic of Korea) to
identify any correlation with the measured coagulation times.
Students t-tests were performed for statistical analysis and
p-value <0.05 represents a statistically significant difference.
Prior to ex vivo tissue experiments, a HIFU transducer was
evaluated in terms of spatial distribution and intensity of acoustic fields. A 0.3-mm (in diameter) needle hydrophone (Precision
Acoustics, Dorset, United Kingdom) was positioned at the
center of the HIFU transducer to measure pressure waveforms.
The output signals were recorded and converted into acoustic
parameters (i.e., acoustic pressure and intensity). The free-field
acoustic pressure at the focal point measured by the hydrophone
was calculated as follows:
pt
Vt
;
(1)
where pt (MPa) is the temporal acoustic pressure, Vt (mV)

is the voltage generated by the acoustic pressure incident on the
active element of the hydrophone, and (mVMPa) is the
hydrophone sensitivity at the acoustic working frequency.
The acoustic intensity at the focal point was then estimated
as follows:
P2eff
;
c
(2)
where Peff is the effective value of pt. In addition to the hydrophone measurements, a Schlieren system was employed to
qualitatively visualize the two-dimensional (2-D) beam shape
of the HIFU transducer near the focal point and to compare
it with the results obtained by the needle hydrophone. The
detailed experimental setup of Schlieren imaging has been introduced elsewhere.25
2.3
Photoacoustic Imaging
To validate coagulated lesions after thermal treatment with

HIFU, all the treated samples were imaged with a PAI system.
Figure 1 illustrates a schematic diagram of the PAI system used
for 2-D imaging. As a light source, a tunable ( 680 to
2500 nm) OPO laser (Surlite OPO Plus, San Jose, California)
was used and pumped by a Q-switched Nd:YAG (Surelite II,
Continuum, San Jose, California) laser with a pulse duration
of 5 ns at 10 Hz. In brief, the laser light from OPO was
perpendicularly reflected at the prism and then spread through

a spherical conical lens, resulting in the ring-shaped pattern. The
ring-shaped light was propagated through a custom-built optical
condenser and was focused onto an artery sample. Upon laser
irradiation, each specimen induced the PA signals, which were
detected by a spherically focused single-element 5.0 MHz ultrasound transducer (V308, Panametrics, Waltham, Massachusetts)
with a focal length of 2.54 cm. In order to reduce the attenuation
of acoustic fields, a targeted sample was immerged in a degassed
water container, and undesirable air bubbles were removed
before conducting PAI. The laser beam was coaligned with
the focal spot of an imaging ultrasound transducer in degassed
water, and for 2-D imaging, each sample in the tissue holder was
moved along x- and y-directions on a 2-D translation stage. The
received PA signals were filtered and amplified by a low-noise
amplifier (5072 PR, Olympus, Waltham, Massachusetts), which
in turn, were converted into digital signals and recorded by
a digital oscilloscope (TDS 5040, Tektronix, Beaverton,
Oregon). The recorded data were used to reconstruct 2-D images
of the HIFU-treated artery samples. The axial and transverse
resolutions of the current PAI system were 144 and 590 m,
respectively. All the reconstructed images were used to estimate
image contrast, defined as the difference between the color of
the coagulated area and its background. Thus, the contrast was
determined by fractional variations in the mean signal amplitudes extracted from regions of interest (ROI) as follows:26
Contrast
CB A
BG
A
BG ;
A
(3)
CB and A
BG are the mean signal amplitudes from the
where A
coagulated blood and adjacent background, respectively. Due
to functional limitations of the current laser system, near-IR
wavelengths (i.e., from 700 to 900 nm with an increment of
50 nm) were merely employed for PAI and tested on the thermally treated samples to identify the spectroscopic effects of
wavelength on image reconstruction and tissue differentiation.
Results
Figure 2(a) illustrates characterization of the focused ultrasound

waves produced by an HIFU transducer. Both axial (blue line)
and longitudinal (red line) distributions of the normalized acoustic field were measured with a hydrophone as a function of radial
distance. According to Fig. 3(a), the transducer generated a focal
spot in an elliptical shape (i.e., 10 mm in axial and 2 mm in
lateral). The Schlieren image [i.e., inlet in Fig. 2(a)] also confirmed the elliptical HIFU beam at the focus shown as an intersecting point between two white dotted lines. In the Schlieren
image, vertical and horizontal lines represented the axial and
longitudinal axes, respectively. The corresponding acoustic
intensity of the HIFU transducer was also estimated and plotted
as a function of acoustic pressure used in the current experiments [Fig. 2(b)]. The intensity increased almost linearly
with the applied pressure, ranging from 1100 to 4600 Wcm2 .
Figure 3 represents the temporal response of complete thermal hemostasis with HIFU in phantom and animal artery samples. Both coagulation time and peak temperature rise were
measured with the phantom samples at various acoustic intensities [Fig. 3(a)]. HIFU exposure was implemented on a total of
25 samples (N 5 per acoustic intensity), and all of the bleeders successfully experienced complete hemostasis. At the lowest
acoustic intensity of 1100 Wcm2 , the thermal coagulation time
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Fig. 2 Characterization of focused ultrasound waves: (a) normalized acoustic pressure in axial (blue)/
longitudinal (red) axes and corresponding Schlieren image (inlet) and (b) acoustic intensity as function of
acoustic pressure.
Fig. 3 Quantitative evaluations on HIFU treatment on targeted artery: (a) coagulation time (red) and
temperature rise (black) from phantom artery at various acoustic intensities and (b) comparison of
coagulation time between phantom and animal arteries (I 3600 Wcm2 ).
was estimated to be almost 2 min and substantially decreased

with increasing intensity down to 30 s. On the other hand,
the temperature rise (T) was initially around 16 K and linearly
increased up to 38 K (at 3600 Wcm2 ), becoming almost saturated afterwards (i.e., T 40 K; p-value 0.35). Figure 3(b)
compares the complete coagulation times between phantom and
Fig. 4 Histological images of animal artery (H&E staining; 100):

(a) control tissue and (b) HIFU-treated animal artery. Note that a
black dotted circle indicates the position of punctured artery sealed
by thermally-induced coagulum (52 s at 3600 Wcm2 ).
animal blood vessels at the acoustic intensity of 3600 Wcm2 .

The percent difference between the two samples was 24%,
indicating that almost 160% slower hemostasis occurred with
the animal sample in comparison with the phantom (i.e., 40
10 s for phantom artery versus 64 20 s for animal artery).
In order to explore the efficacy of HIFU treatment on animal
artery, histological analysis stained with H&E was performed as
shown in Fig. 4. A control image [Fig. 4(a)] shows a cross-sectional area of the animal blood vessel without any punctured
lesion. Figure 4(b) demonstrates that the puncture was initially
made at 7 oclock in the blood vessel area and thermally treated
with HIFU at 3600 Wcm2 for 52 s. From the region of interest,
the punctured vessel wall was sealed by coagulum, which
extruded through the media and adventitia and partially covered
the outer surface of the artery. Thus, the histology image in
Fig. 4(b) confirmed that the complete hemostasis was achieved
with the HIFU thermal treatment.
Figure 5 exhibits top-view images of coagulated artery samples after HIFU treatment. A dotted circle indicated the position
of the HIFU-treated area on the phantom artery at the acoustic
intensity of 3600 Wcm2 for 40 s [Fig. 5(a)]. The incision hole
was sealed with blood coagulum (i.e., black color inside the
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Fig. 5 PAI mapping of tissues after HIFU coagulation: photographs of (a) phantom and (c) animal
arteries (I 3600 Wcm2 ) and corresponding PA images of (b) phantom and (d) animal arteries
acquired at 700 nm wavelength.
sample) and whitish surface discoloration (i.e., on the sample

surface) after thermal treatment, which eventually prevented
additional bleeding from the punctured sample. Figure 5(b)
presents a PA image of the same sample in Fig. 5(a) taken
by a PAI system ( 700 nm) after the complete hemostasis.
The coagulated region in a white dotted circle demonstrated
a two times higher contrast in the amplitude of PA signals as
the untreated region along with a signal-to-noise ratio (SNR) of
4 dB. It was confirmed that the thickness of the sample in
Fig. 5(b) was equivalent to that of the sample in Fig. 5(a).
Figure 5(c) shows the HIFU-treated animal artery at the acoustic
intensity of 3600 Wcm2 for 52 s. The lesion (i.e., 1 mm in
diameter) in a black dotted circle evidenced the complete
thermal hemostasis, covering the artery surface with black
coagulum. Based upon the acquired images, the extent of

coagulative necrosis in the peripheral area was found to be
0.5 0.2 mm. In Fig. 5(d), a PA image of the same animal
artery was taken at the 700-nm wavelength. Similarly to
Fig. 5(b), the coagulated area showed distinctively higher contrast in the blood vessel image. Both sample images in Figs. 5(c)
and 5(d) represented the equivalent sample thickness of 2 mm.
To quantitatively characterize a PA image of HIFU-treated
animal artery in Fig. 5(d), the PA maximum amplitude projection at three different locations in the image was measured and
compared for characteristics such as background, blood vessel,
and coagulated regions as shown in Fig. 6(a) (i.e., 0.228 0.031
for coagulated region versus 0.006 0.001 for background and
0.095 0.013 for blood vessel). Due to significant contrast
Fig. 6 Quantitative characterization of PA image: (a) comparison of PA image contrasts acquired at

700 nm and (b) spectroscopic evaluation on PA signal amplitudes.
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variations, PAI was able to differentiate the HIFU-induced

coagulation area from background and other untreated areas.
The image contrast from the coagulated region showed approximately 37 and 1.4 times higher than those from the background
and blood vessel, respectively. To further identify the optimal
PAI wavelength for precisely detecting thermally coagulated
lesions after HIFU treatment, the PA amplitudes of the coagulated areas in the animal arteries were measured and compared at
various incident laser wavelengths (from 700 to 900 nm by
50 nm increment). All the tissue samples were treated at the
acoustic intensity of 3600 Wcm2 for 52 s. Apparently, the
spectroscopic measurements showed that the wavelength of
700 nm achieved the maximum light absorption by the coagulated area [Fig. 6(b)] in comparison with other wavelengths.
Beyond 700 nm, the PA amplitude significantly decreased
with the wavelength.
Discussion
The current study presented the feasibility of a noninvasive

HIFU therapeutic method for complete hemostasis in both phantom and animal arteries. Typically, blood coagulation requires
a temperature increase up to 343 K induced by physical
interactions of acoustic energy with soft tissues.8 Similarly,
high-acoustic intensity rapidly localized and increased the temperature (up to T 38 K within 30 s at 4600 Wcm2 ) within
the focal point in tissue as shown in Fig. 3(a). Thus, the relatively faster temperature development eventually resulted in
more efficient hemostasis in the phantom and animal blood vessels. Evidently, the temperature increase is linearly associated
with the applied acoustic intensity as well as tissue properties
(acoustic and thermal) as follows:27
2
I;
c
(4)
where dBm is the acoustic absorption coefficient at the

center frequency. Then, given the properties for the blood sample (i.e., 1060 kgcm3 and 3 dBm),28 the estimated
temperature rises at 1100 and 1700 Wcm2 can be 17 and
26 K, respectively; accordingly, the theoretical values showed
a good agreement with the temperature increase measured in
the current study as shown in Fig. 3(a). However, at the higher
acoustic intensities (i.e., 3600 and 4600 Wcm2 ), the temperature increase became saturated at T 40 K. The saturation
behavior could be primarily associated with the thermal insulation effect of the solidified coagulum on the punctured site.
Once thermal denaturation was initiated, coagulum could
begin to form throughout the phase change and eventually
cover the bleeding area. Upon complete hemostasis, the most
thermal energy would be deposited in the generated coagulum,
entailing the formation of multiple microbubbles on the target
surface, which was reported by Clarke et al.29 According to their
study, the aggregation of microbubbles would result in the formation of an acoustic barrier to the ultrasound beam on the target surface. Then, most of the incident acoustic energy could be
scattered and/or partially absorbed by the bubble layer to entail
inefficient energy coupling. As the temperature underneath the
barrier would start to decrease due to the increased attenuation,
a thermocouple would be able to merely measure the temperature change beneath the coagulated area up to T 40 K. Thus,
it is conceived that the saturation behavior occurred due to the
consequent temperature decrease by the surface bubble barrier
during HIFU thermal therapy. In fact, the current study was able
to observe the generation of a group of macrobubbles on the
coagulum surface during the HIFU treatment. Accordingly,
the acoustic intensity of 3600 Wcm2 might be the threshold
intensity to obtain complete hemostasis under the current conditions. Further studies will perform temperature measurements
at various points in tissue to map the spatial distribution of thermal energy and to identify the optimal acoustic intensity for
rapid coagulation.
Post-experimental measurements confirmed the significant
extent of collateral injury of 0.5 0.2 mm. It is conceivable
that the thermal injury could result from both spatial and temporal effects of HIFU treatments. Compared to the diameter of
the punctured hole, the HIFU beam was relatively larger (i.e., 10
by 2 mm in Fig. 2), which could have covered even the healthy
tissue regions. However, there was a trade-off between collateral
damage and completion of hemostasis. According to our preliminary studies, complete coagulation was difficult to achieve
with the smaller size of the HIFU beam due to higher light
intensity along with concentrated temperature increase as well
as limited beam alignments. Particularly, the accurate positioning of the smaller beam size on the targeted tissue area
extensively prolonged the entire testing time. Thus, the wider
distribution of the HIFU beam was more applicable and practical to readily accomplish complete hemostasis as well as fast
beam alignments in spite of the inevitably considerable thermal
injury.
The current study found that the complete hemostasis
required a significant amount of treatment time (i.e., 64 20 s)
for animal tissue. Based upon the treatment time, the spatial
extent of heat diffusion could roughly be calculated to be 3 mm
by using the following equation:30
ztherm
p
4 t;
(5)
where ztherm (m) is the time-dependent thermal penetration

depth, m2 s is the thermal diffusivity (e.g., 14 107 m2 s
for soft tissue), and t (s) is the heat accumulation time. However,
it should be noted that the theoretical depth was six times thicker
than that measured in the current study (i.e., 0.5 0.2 mm).
The thinner coagulative necrosis was conceivably associated
with convective heat transfer due to blood perfusion at 20 mlh
and the degased water environment maintained at 38C. Vaezy
et al. reported that a significant amount of heat was carried
downstream during HIFU application, consequently protecting
the blood vessel wall from thermal injury and simultaneously
protracting the thermal treatment time to reach the targeted
temperature.31 Therefore, additional quantitative investigations
with animal tissue are still necessary to identify the optimal
treatment dose and conditions in light of beam size, coagulation
time, and minimal degree of thermal injury. In an effort to evaluate the thermal treatment time, t (s), required for complete
hemostasis of tissue, the Arrhenius rate equation was also used
as follows:32
t eT c T 0 T c ;
(6)
where is the Arrhenius integral, is the rate constant (i.e.,

244.8 for blood flow), T c is the critical temperature (i.e.,
327.7 K for microvascular blood flow), and T 0 is the coagulation temperature (i.e., 323.15 K).32 Since thermal coagulation is
associated with irreversible conformational changes in blood
(i.e., 1), the estimated heating interval can be calculated
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to be 62.4 s, which is comparable to the experimentally measured time (i.e., 64 20 s). However, the current study defined
the coagulation time as the moment when hemostasis was
achieved through visual inspection. Both post-experimental
histology analysis and PAI imaging merely confirmed coagulation with 1. Accordingly, further information on the temperaturetime relation during coagulation will be required in
order to precisely assess the initiation of coagulation events
and to validate the end-point of HIFU treatment.
Figure 3(b) exhibited that the coagulation time measured for
animal artery was 160% longer than that for the phantom one at
3600 Wcm2 (i.e., 64 20 s for animal artery versus 40 10 s
for phantom artery). It should be noted that the phantom artery
made of silicon has an order of Youngs modulus higher
than that of animal tissue (i.e., 2.07 MPa for phantom artery33
versus 0.19 MPa for animal tissue).34 In fact, the hole in the
phantom artery was often deformed and became a <0.5-mm
wide, elongated cut whereas the animal artery almost maintained the original shape of the needle-drilled hole. Accordingly,
the geometrically altered hole in the phantom artery could have
been instrumental in facilitating the thermal coagulation process
during HIFU treatment. Moreover, since the wall of the animal
blood vessel was less uniform and thinner than that of the phantom tube (i.e., 1 mm for phantom artery versus 0.5 mm for
animal artery), structural differences could have contributed
to the longer coagulation time owing to the relatively shorter
pathway of coagulation along the incised hole.
The acquired PA images in Fig. 5 demonstrated that thermal
lesions were well defined with higher contrast, compared to the
surrounding areas. Image contrast typically represents the
amplitude of the acquired PA signal, which is related to the optical energy locally absorbed by tissue chromophores (i.e., oxygenated hemoglobin of blood and water). Thus, the enhanced
contrast indicated strong light absorption by the volumetric tissue specifically exposed to HIFU (Fig. 5). Besides, the increased
PA amplitudes acquired from the HIFU-treated lesions agreed
well with the findings from the previous study.35 It was reported
that the thermomechanical properties of tissue, expressed as a
Grneisen coefficient, could contribute to augment PA signals
in coagulated tissue, in that the coefficient for coagulated blood
was 65% higher than that for the native sample (i.e., 0.06 for
noncoagulated and 0.09 for coagulated blood).36 The Grneisen
coefficient, , is related to the initial acoustic pressure, p0 , upon
light absorption, which can be expressed as p0 a ,
where a cm1 and Jcm2 denote optical absorption
coefficient and light fluence, respectively.37 Black et al. reported
that the light absorption coefficient of clotted blood was higher
than one in the native state (i.e., a 8 cm1 for clotted blood
versus 1 cm1 for native blood at 700 nm).38 Under the same
fluence, it is conceivable that both Grneisen and absorption
coefficients can primarily determine the degree of acoustic transients as well as the quality of PA imaging. In fact, the bright
areas in the acquired PA images were validated to correspond to
local accumulation of solid coagulum after HIFU-induced denaturation (Fig. 5). Accordingly, the coagulated blood with the
higher Grneisen as well as optical absorption coefficients
could promote stronger acoustic transients and result in more
distinctive image contrast.
Spectroscopic studies on PA signal amplitudes confirmed
that the degree of light absorption by thermally treated tissue
at shorter optical wavelengths was higher than that at longer
wavelengths due to relatively stronger PA amplitudes, which
Fig. 7 Schematic diagram of HIFU thermal treatment integrated

with fiber-based PAI for real-time monitoring.
was consistent with ONeills report. Their study showed that

the normalized PA intensity at HIFU-treated bovine muscle
was 2.5 times strong at shorter wavelengths than longer wavelengths (i.e., 0.4 at 700 nm versus 0.16 at 900 nm).39 It
was noted that the absorption coefficients of bovine muscle
were similar to those of deoxyhemoglobin between 700 and
900 nm.40,41 Thus, coagulated blood could strongly absorb
the incident laser light, consequently entailing higher PA contrast in reconstructed images. Furthermore, the spectroscopic
PA results demonstrated the strongest PA signal amplitude measured at 700 nm [Fig. 7(b)], which possibly attributed to more
light absorption specifically by the coagulated tissue. Therefore,
the stronger acoustic contrast at 700 nm led up to a 90% higher
SNR of 14.4 dB between coagulated blood and background,
whereas 11.4, 7.9, 9.2, and 11.8 dB were estimated at 750,
800, 850, and 900 nm, respectively.
Although the current study successfully demonstrated the
PAI detection of thermal lesions induced by HIFU coagulation,
experimental limitations still remain for the sake of practical
application. During the experiments, the coagulated areas for
hemostasis were imaged with PAI followed by HIFU application
due to complexities of the current system. Thus, the 2-D images
hardly contained any temporal information for exhibiting the
thermal coagulation process as well as capturing the moment
for the complete hemostasis. A thermocouple was also used
and inserted to measure the maximum temperature in the treated
tissue (Fig. 1). Thus, in-depth considerations upon the integrated
probes would be critical for real-time monitoring thermal treatment and precluding any postbleeding events. In fact, PAI
assessment of blood coagulation can be feasible only if all
the signal measurements are performed real-time and coaxially
with HIFU treatment. Figure 7 shows a schematic diagram of
fiber-based PAI transducers that is coaxially integrated with
the HIFU transducer. Due to coalignment with the equivalent
focal length, the designed transducer would readily identify
the treated area as well as capture PA signals in situ.
Presently, the follow-up studies with the combined probes are
underway to identify the optimal treatment dosage, to detect
the PA signals during/after HIFU treatment, to calibrate 1-D
information with temperature elevation, and to acquire 2-D
images as a validation tool for complete hemostasis.24 The
change and rate of change in PAI amplitudes will specifically
be assessed and correlated to visual observation of hemostasis
and histological outcome. In turn, the temperature-time relation
of coagulation with PAI can reliably elucidate the relevance
of PAI in determining thermal damage and its relationship to
105010-7
October 2014
Vol. 19(10)
hemostasis, given the data of blood and blood vessel for

the Arrhenius integral. In addition, the current testing utilized
heparinized blood for the purpose of easy sample maintenance,
which might adversely have protracted the time required to
achieve the complete hemostasis possibly by 2 to 10 times
longer, in comparison with a nonheparinized blood sample.8
Furthermore, the current study used transparent media (i.e.,
degased pure water) for PAI to minimize any optical scattering
effects that could accompany weak signal generation with low
image contrast, lateral distribution of photons, and shallow
depth for volumetric imaging. Thus, in vivo dorsal chambers in
a rat model will be used to examine any biochemical and
optical scattering effects of vascularized samples on the
coagulation process as well as image reconstruction at various
wavelengths. Finally, the ultimate research goal is to develop
a portable image-guided HIFU system for rapid hemostasis
with minimal collateral damage to tissue. Further investigations will continue to focus on design and development of a
rechargeable MOSFET-based RF amplifier with a hand-held
HIFU transducer that can be compatible with an optical or
ultrasound imaging system.
Conclusion
The current study presented the feasible application of HFU for

thermal hemostasis assisted by visualization of the nonionizing
PAI modality under ex vivo conditions. Acoustic energy was
successfully applied to achieve complete hemostasis in artery
along with minimal damage to the peripheral tissue. PAI was
able to provide information on the location of HIFU-treated
lesions, indicating the great potential to provide feedback on
HIFU thermal treatment. The systematic integration of PAI
and HIFU will be conducted to achieve the real-time monitoring
of the hemostasis process on the injured blood vessels in vivo.
The proposed technique can be a feasible tool to facilitate
blood coagulation and ensure the safety of HIFU treatment with
a portable system.
Acknowledgments
This research was supported by Basic Science Research
Program through the National Research Foundation of Korea
(NRF) funded by the Ministry of Education, Science and
Technology (NRF-2012R1A1A1012965). The authors would
like to appreciate Mr. Trung Hau Nguyen for his help on
discussion.
References
1. J. K. Barton, D. P. Popok, and J. F. Black, Thermal analysis of
blood undergoing laser photocoagulation, IEEE J. Sel. Top. Quantum
Electron. 7(6), 936943 (2001).
2. R. R. Krueger and E. E. Almquist, Argon laser coagulation of blood
for the anastomosis of small vessels, Lasers Surg. Med. 5(1), 5560
(1985).
3. R. E. Rumbaut, Platelet-vessel wall interactions in hemostasis and
thrombosis, Morgan & Claypool Publishers LLC, San Rafael,
California (2010).
4. S. Vaezy et al., Liver hemostasis with high-intensity ultrasound: repair
and healing, J. Ultrasound Med. 23(2), 217225 (2004).
5. S. Vaezy and V. Zderic, Hemorrhage control using high intensity
focused ultrasound, Int. J. Hyperthermia 23(2), 203211 (2007).
6. U. Maestroni et al., High-intensity focused ultrasound for prostate
cancer: long-term follow up and complications rate, Adv. Urol.
2012, 14 (2012).
7. R. Ramanathan and R. J. Leveillee, Ablative therapies for renal

tumors, Ther. Adv. Urol. 2(2), 5168 (2010).
8. S. Vaezy et al., Use of high-intensity focused ultrasound to control
bleeding, J. Vasc. Surg. 29(3), 533542 (1999).
9. Y. Zhou, Generation of uniform lesions in high intensity focused
ultrasound ablation, Ultrasonics 53(2), 495505 (2013).
10. J. E. Kennedy, High-intensity focused ultrasound in the treatment of
solid tumours, Nat. Rev. 5(4), 321327 (2005).
11. E. R. Cordeiro et al., High-intensity focused ultrasound (HIFU) for
definitive treatment of prostate cancer, BJU Int. 110(9), 12281242
(2012).
12. A. Burgess et al., High-intensity focused ultrasound (HIFU) for dissolution of clots in a rabbit model of embolic stroke, PLoS One 7(8),
e42311 (2012).
13. C. Wright, K. Hynynen, and D. Goertz, In vitro and in vivo high-intensity focused ultrasound thrombolysis, Investig. Radiol. 47(4), 217225
(2012).
14. M. R. Pfeffer et al., Image-guided high-intensity focused ultrasound in
the treatment of cancer, in Image-Guided Cancer Therapy, D. E.
Dupuy, Y. Fong, and W. N. McMullen, Eds., pp. 7999, Springer,
New York (2013).
15. B. E. ONeill et al., MRI-based prediction of pulsed high-intensity
focused ultrasound effect on tissue transport in rabbit muscle,
J. Magn. Reson. Imaging 38(5), 10941102 (2013).
16. R. W. Martin et al., Hemostasis of punctured vessels using Dopplerguided high-intensity ultrasound, Ultrasound Med. Biol. 25(6), 985
990 (1999).
17. S. Vaezy et al., Real-time visualization of high-intensity focused ultrasound treatment using ultrasound imaging, Ultrasound Med. Biol.
27(1), 3342 (2001).
18. C. L. Bayer, G. P. Luke, and S. Y. Emelianov, Photoacoustic imaging
for medical diagnostics, Acoustics Today 8(4), 1523 (2012).
19. S. Mallidi, G. P. Luke, and S. Emelianov, Photoacoustic imaging in
cancer detection, diagnosis, and treatment guidance, Trends Biotechnol.
29(5), 213221 (2011).
20. C. Kim, C. Favazza, and L. V. Wang, In vivo photoacoustic tomography of chemicals: high-resolution functional and molecular optical
imaging at new depths, Chem. Rev. 110(5), 27562782 (2010).
21. C. Kim et al., In vivo molecular photoacoustic tomography of melanomas targeted by bioconjugated gold nanocages, ACS Nano 4(8),
45594564 (2010).
22. G. Ku and L. V. Wang, Deeply penetrating photoacoustic tomography
in biological tissues enhanced with an optical contrast agent, Opt. Lett.
30(5), 507509 (2005).
23. K. H. Song and L. V. Wang, Deep reflection-mode photoacoustic
imaging of biological tissue, J. Biomed. Opt. 12(6), 060503 (2007).
24. P. V. Chitnis et al., Feasibility of optoacoustic visualization of highintensity focused ultrasound-induced thermal lesions in live tissue,
J. Biomed. Opt. 15(2), 021313 (2010).
25. T. Neumann and H. Ermert, Schlieren visualization of ultrasonic wave
fields with high spatial resolution, Ultrasonics 44(Suppl. 0), e1561
e1566 (2006).
26. J. L. Su et al., Photoacoustic imaging of prostate brachytherapy seeds,
Biomed. Opt. Express 2(8), 22432254 (2011).
27. L. Faqi et al., Measuring temperature rise in phantom to determine
high power high-intensity focused ultrasound sound field, in Fourth
Int. Conf. Bioinformatics and Biomedical Engineering, pp. 14
(2010).
28. M. Solovchuk, T. W. Sheu, and M. Thiriet, Simulation of nonlinear
Westervelt equation for the investigation of acoustic streaming and
nonlinear propagation effects, J. Acoust. Soc. Am. 134(5), 39313942
(2013).
29. R. L. Clarke and G. R. ter Haar, Temperature rise recorded during
lesion formation by high-intensity focused ultrasound, Ultrasound
Med. Biol. 23(2), 299306 (1997).
30. M. H. Niemz, Laser-Tissue Interactions Fundamental and
Applications, 3rd ed., Springer_Verlag, Berlin Heidelberg, New
York (1996).
31. S. Vaezy et al., Hemostasis using high intensity focused ultrasound,
Eur. J. Ultrasound 9(1), 7987 (1999).
32. http://fieldp.com/myblog/2008/arrhenius-rate-integrals-in-computerthermal-solutions/.
105010-8
October 2014
Vol. 19(10)
33. N. I. Inc., http://www.newageindustries.com/downloads/catalogs/

NewAge_Silicone_Catalog.pdf.
34. T. Khamdaeng et al., Arterial stiffness identification of the human
carotid artery using the stress-strain relationship in vivo, Ultrasonics
52(3), 402411 (2012).
35. I. V. Larina, K. V. Larin, and R. O. Esenaliev, Real-time optoacoustic
monitoring of temperature in tissues, J. Phys. D 38(15), 26332639
(2005).
36. B. Soroushian, W. M. Whelan, and M. C. Kolios, Assessment of
opto-mechanical behavior of biological samples by interferometry,
Proc. SPIE 7177, 71771X (2009).
37. B. Cox et al., Quantitative spectroscopic photoacoustic imaging:
a review, J. Biomed. Opt. 17(6), 061202 (2012).
38. J. K. Barton et al., Cooperative phenomena in two-pulse, two-color
laser photocoagulation of cutaneous blood vessels, Photochem.
Photobiol. 73(6), 642650 (2001).
39. Y. Sun and B. ONeill, Imaging high-intensity focused ultrasoundinduced tissue denaturation by multispectral photoacoustic method:
an ex vivo study, Appl. Opt. 52(8), 17641770 (2013).
40. W. G. Zijlstra, A. Buursma, and W. P. Meeuwsen-van der Roest,

Absorption spectra of human fetal and adult oxyhemoglobin, de-oxyhemoglobin, carboxyhemoglobin, and methemoglobin, Clin. Chem.
37(9), 16331638 (1991).
41. J. J. Xia et al., Characterizing beef muscles with optical scattering and
absorption coefficients in VIS-NIR region, Meat. Sci. 75(1), 7883
(2007).
Van Phuc Nguyen received the masters degree in mechatronics
engineering from Pukyong National University, Busan, Republic of
Korea, in 2012. He is currently working as a PhD candidate in the
Bio-Therapeutics Laboratory under the supervision of professor
Hyun Wook Kang. His research interests are developing an integration of high intensity focused ultrasound (HIFU) and photoacoustic
imaging system in real-time and mobile applications.
Biographies of the other authors are not available.
105010-9
October 2014
Vol. 19(10)

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