Ultrasound in Med. & Biol., Vol. 34, No. 8, pp.

1281–1291, 2008
Copyright © 2008 World Federation for Ultrasound in Medicine & Biology
Printed in the USA. All rights reserved
0301-5629/08/$–see front matter

doi:10.1016/j.ultrasmedbio.2007.12.020

● Original Contribution

EFFECTS OF ACOUSTIC INSONATION PARAMETERS ON

ULTRASOUND CONTRAST AGENT DESTRUCTION

CHIH-KUANG YEH and SHIN-YUAN SU

Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University,
Hsinchu, Taiwan

(Received 8 April 2007, revised 15 December 2007, in final form 21 December 2007)

Abstract—Ultrasound contrast agents (UCAs) are used to enhance the acoustic backscattered intensity of blood
and thereby assist the assessment of blood perfusion. Characterization of UCA destruction provides important
information for the design of contrast-assisted perfusion imaging. High-speed optical observation of single
microbubble destruction during acoustic insonation has been performed in previous studies. The results
identified that pressure, center frequency and transmission phase have significant effects on the fragmentation
threshold. We proposed an acoustic-based experiment method to demonstrate the relationship between different
acoustic exposure conditions and the degree of UCA destruction. The method also provides a simple and
convenient way to determine the microbubble destruction threshold. The experiments introduced three in-
sonation parameters, including acoustic pressure (0 to 1 MPa), pulse frequency (1, 2.25, 5 and 7.5 MHz) and pulse
length (1 to 10 cycles). The term of surviving percentage (SP) was proposed to represent the ratio of UCA
backscattered power with and without acoustic insonation. The results showed that the SP decreased with
decreasing pulse frequency, but with increasing transmission acoustic pressure and pulse length. In addition,
there was an exponential relationship between SP and acoustic pressure, and thus the UCA destruction pressure
threshold could be predicted from the fitted exponential curve. The results also show that the degree of UCA
destruction was not related to mechanical index (MI). Potential applications of this method include UCA
high-resolution destruction/replenishment imaging model, microbubble cavitation, sonoporation in drug delivery
and gene therapy. (E-mail: ckyeh@mx.nthu.edu.tw) © 2008 World Federation for Ultrasound in Medicine &
Biology.

Key Words: Ultrasound contrast agents, Surviving percentage, Mechanical index.

INTRODUCTION mitted pulse. The nonlinear UCA imaging modalities

Ultrasound contrast agents (UCAs) are used to enhance
echoes backscattered from blood and thereby assist mea-
surements of blood flow. UCAs are typically shell-en-
capsulated microbubbles whose solubility in the blood is
sufficiently low to ensure that they reach the left ventri-
cle of the heart without significant losses. One of the
most important characteristics of UCAs is their nonlinear
oscillation, which has been investigated in many exper-
imental and theoretical studies (Chang et al. 1995; Fors-
berg et al. 2000; Goldberg et al. 2001; Lotsberg et al.
1996; Shi et al. 1999, 2000). Nonlinear oscillations allow
tissue and UCA echoes to be distinguished based on their
center frequency or response to the phase of the trans-
Address correspondence to: Chih-Kuang Yeh, Ph.D., Depart-
ment of Biomedical Engineering and Environmental Sciences, National
Tsing Hua University, 101, Section 2, Kuang-Fu Road, Hsinchu, Tai-
wan 30013, ROC. E-mail: ckyeh@mx.nthu.edu.tw
developed to date include phase-inversion, subharmonic
and ultraharmonic methods (Lotsberg et al. 1996; Shi et
al. 1999; Simpson et al. 1999).
Another important characteristic of UCAs is that
they can be fragmented by suitable acoustic excitation.
Expanding the microbubble radius on the order of 300%
will cause the microbubble to collapse rapidly and sep-
arate into small fragments (Bouakaz et al. 2005; Chen et
al. 2002; Chomas et al. 2001a). It is well known that
UCAs can be destroyed reliably using ultrasound pulses
with center frequencies ⬍5 MHz, and Chomas et al.
(2001b) found that decreasing the center frequency re-
duces the threshold pressure required for microbubble
destruction. Some studies have applied the fragmentation
property of UCAs to estimate blood perfusion. Wei et al.
(1998) proposed a well-known blood perfusion evalua-
tion model, the microbubble destruction/replenishment
technique, in which contrast agents are destroyed by

1281
1282 Ultrasound in Medicine and Biology
excitation with lower-frequency pulses, and the subse-
quent flow of intact UCAs into the sample volume is
monitored over time to produce a local estimate of the
blood flow velocity. This model has been used to study
UCA replenishment in rat tumors (Chomas et al. 2001c)
and other disease models (Wei et al. 2001; Yeh et al.
2004).
The UCA destruction process can also be exploited
to improve the contrast between blood and tissue in
images. Frinking et al. (2001) suggested applying a re-
lease burst to rupture the microbubbles, and then distin-
guishing the microbubbles from the background tissue
by measuring the decorrelation between the echo signals
before and after the excitation. Furthermore, the destruc-
tion process provides potential application in acoustic
cavitation as well as new techniques for drug delivery
and gene therapy (Bloch et al. 2004a).
Theoretical and experimental methods have been
developed for investigating UCA fragmentation (Bloch
et al. 2004b; Chomas et al. 2001a; Porter et al. 2006;
Smith et al. 2007). Characterizing microbubble destruc-
tion provides important information for the design of
contrast-assisted perfusion imaging systems. The dy-
namic motion of microbubbles during acoustic in-
sonation has been observed optically (Bouakaz et al.
2005; Chomas et al. 2000, 2001b). Images captured by a
high-speed camera system have shown the destruction of
single microbubbles during the ultrasound compression
phase. The experimental results suggested that the rest-
ing microbubble radius, acoustic excitation pressure,
pulse center frequency and pulse length determine the
probability of fragmentation. Increasing the pressure and
the pulse length, and decreasing the resting radius and
pulse frequency are correlated with an increased proba-
bility of UCA fragmentation.
Commercial UCAs are generally delivered through
a patient IV and circulated through the body. Because a
large number of microbubbles pass simultaneously
through the imaging sample volume, the threshold for
microbubble destruction will be affected by variability in
bubble scattering and shadowing effects (Yeh et al.
2003). Thus, an index is needed that accurately predicts
the degree of microbubble destruction induced by expo-
sure to a diagnostic ultrasound field. In this study, we
used an acoustic-based experimental method and an ex-
ponential model to predict the degree of microbubble
destruction. The method provides a simple way to deter-
mine the microbubble destruction threshold in common
laboratory experiments. Different acoustic exposure con-
ditions were considered under clinically realistic diag-
nostic ultrasound fields, including pulse center frequen-
cies from 1–7.5 MHz, acoustic excitation pressures up to
1 MPa and pulse lengths from 1–10 cycles.
Volume 34, Number 8, 2008
Most clinical investigators consider the value of the
mechanical index (MI) to be an indicator of exposure
conditions related to UCA destruction (Forsberg et al.
2005; Hirokawa et al. 2002). The MI is proportional to
the peak negative pressure of the excitation pulse, and
increasing the pulse pressure increases the probability of
microbubble fragmentation. However, minimizing the
excitation pulse pressure required to induce fragmenta-
tion is desirable in clinical applications. Forsberg et al.
(2005) evaluated the MI as a predictor of exposure con-
ditions related to the destruction of UCAs in B-mode
images obtained using a clinical scanner. They con-
cluded that the MI index does not predict the degree of
microbubble destruction and hence should not be used by
itself to define the exposure conditions for destroying
microbubble contrast agents. Because the MI relates to a
nonthermal mechanism that may produce bioeffects
(O’Brien 2007), we also investigated the relationship
between the MI and the degree of microbubble destruc-
tion in this study.
This paper is organized as follows. We first describe
the setup for the acoustic-based experiments used to
estimate the degree of microbubble destruction. Then,
the microbubble destruction results under different
acoustic exposure conditions are presented. An exponen-
tial mathematical model is introduced to predict the
relationship between the degree of microbubble destruc-
tion and the acoustic pressure. The relationship between
the MI and the degree of microbubble destruction is
discussed and, finally, conclusions are drawn.
MATERIALS AND METHODS
Experimental setup
An acoustical experimental setup was designed to
determine the microbubble destruction threshold under
different insonation conditions. Three acoustic in-
sonation parameters were used: (i) pulse pressures vary-
ing from 0 –1 MPa; (ii) pulse lengths of 1, 3, 5 and 10
cycles; and (iii) pulse center frequencies of 1, 2.25, 5 and
7.5 MHz. Figure 1 shows a block diagram of the exper-
imental system. The setup consists of two types of sin-
gle-element transducers: (i) a 25-MHz spherically-fo-
cused transducer (model V324, Panametrics, Waltham,
MA, USA) responsible for imaging and (ii) four lower-
frequency spherically-focused transducers with center
frequencies of 1, 2.25, 5 and 7.5 MHz used for micro-
bubble destruction. The specifications of the transducers
are summarized in Table 1.
The 25-MHz transducer was fixed at 45 degrees
relative to and near the outlet of a 200-␮m inner diameter
cellulose tube (Spectrum Labs, Laguna Hills, CA, USA),
with the focal region of the transducer aligned onto the
tube. The wall thickness of the tube is 16 ␮m and the
 Microbubbles destruction ● C.-K. YEH and S.-Y. SU 1283

Fig. 1. Experimental arrangement for the threshold of bubble destruction measurements.

attenuation effect in either imaging transducers or de- maximum transmitted pressure of the 1-MHz transducer
struction transducers is negligible. The four low-fre- was 0.7 MPa, and the 7.5-MHz transducer was 0.8 MPa.
quency destruction transducers were fixed approximately The destruction pulses were transmitted with a pulse
5 cm upstream from the 25-MHz transducer on the inlet repetition frequency (PRF) of 50 Hz, and the primary and
side of the tube, perpendicular to the tube, and the focus secondary radiation forces were insignificant and hence
of the destruction transducer was also located inside the could be ignored (Dayton et al. 1997). The radiation
tube. Acoustically absorbent rubber was placed at the force is related to the flow and bubble concentration in
bottom of the water tank to minimize reflections. the acoustic filed. The microbubbles suffer less influence
A digital-to-analog board (model TE5300, Tabor of radiation force under lower PRF insonation pulses. In
Electronics, Tel Hanan, Israel) was used to generate 1-, each combination of frequency, pulse duration and
2.25-, 5- and 7.5-MHz tone-burst pulses comprising 1, 3, acoustic pressure amplitude, there were 10 independent
5 and 10 cycles, and these drove the transducers via a repetitions experiments and the total exposure duration
radiofrequency (RF) power amplifier (model 150A100B, was 6 s.
AR, Souderton, PA, USA). The peak negative pressures After a delay of 5 s after excitation with the lower-
generated by the four destruction transducers were cali- frequency pulses, the 25-MHz transducer was operated
brated to an accuracy of 0.1 MPa using a wideband in a pulse/echo mode to detect the presence of bubbles in
hydrophone (model HNP-0400, ONDA, Sunnyvale, CA, the tube. The signals were received by a pulser/receiver
USA) before performing the experiments. Note that the (model 5900PR, Panametrics) operating in the pulse/
echo mode via a diode limiter/transformer diplexer cir-
cuit at a PRF of 500 Hz. The peak-negative pressure
Table 1. Transducer characteristics level of the 25-MHz transducer was 0.24 MPa at the
focus. The passband of the pulser/receiver hardware filter
Element Focal –6 dB
Model Frequency Size Length Bandwidth (%) was from 10 –50 MHz. The energy of the excitation
imaging pulse was 4 ␮J, and the receiver gain was 26 dB.
V303 1 12.7 18.5 78.2 A preamplifier connected upstream of the pulser/receiver
V304 2.25 25.4 71.3 74.9
V308 5 19.1 72.1 59.4 (model AU-1114-BNC, Miteq, Hauppauge, NY, USA)
V321 7.5 19.1 35.6 94.1 amplified the RF signal by 31 dB, which was then
V324* 25 6.4 12.7 50.6 digitized by a 120-MHz, 14-bit analog-to-digital board
Unless otherwise noted, the unit of measure is in millimeters. (model PCI-9820, ADLINK Technology, Taipei, Tai-
* Imaging transducer. wan) and stored in M-mode (i.e., motion mode) format
1284 Ultrasound in Medicine and Biology Volume 34, Number 8, 2008

on a personal computer (Hewlett-Packard, Palo Alto,

CA, USA). MATLAB (The MathWorks, Natick, MA,
USA) was used to analyze the data off-line. M-mode
format means that each vertical line corresponds to the
received signal along the axial direction. Such signals are
obtained repetitively at a rate defined by the PRF and are
stacked along the horizontal axis.
A commercial contrast agent (Definity, Bristol-My-
ers Squibb Medical Imaging, N. Billerica, MA, USA)
was used at a concentration of 6 ␮L/10 mL water (3
mL/5 L blood volume). Definity microbubbles have di-
ameters ranging from 1.1–3.3 ␮m and a maximum par-
ticle size of about 20 ␮m, with over 98% of them ⬍10
␮m. Note that each microliter of the Definity suspension
contains the 1.2 ⫻ 1010 and 4,800 microspheres before
and after dilution, respectively. The dose of Definity
usually given to human patients is 10 ␮L/kg by IV bolus
injection. A syringe pump maintained the flow velocity
of the UCAs solution through the tube at 8.9 mm/s, Fig. 2. (a) RF backscattered signal. (b) RF M-mode image. (c)
corresponding to a flow rate of 1 mL/h. Postwall-filtered envelop-detected M-mode image. (d) Sum of the
Because the lateral beamwidth of the 25-MHz trans- postwall-filtered power over the time interval from 0–0.6 s in (c).
ducer is approximately 120 ␮m, there were fewer than 15
microbubbles within the sample volume. The backscat-
tered signal power received by the 25-MHz transducer (i.e., – 6dB bandwidth of 19 to 32 MHz) used in this
was proportional to the number of microbubbles within study. In other words, nonlinear excitation is negligible
the sample volume at the pressures (i.e., 0.24 MPa at for the 25-MHz imaging transducer. To consider the
focus), pulse lengths (i.e., 4 to 5 cycles) and frequencies condition of first-pass concentration of microbubbles in

Fig. 3. Postwall-filtered power as a function of acoustic pressure for a 1-MHz transducer with (a) 1-, (b) 3-, (c) 5- and
(d) 10-cycle transmitted pulses.
 Microbubbles destruction ● C.-K. YEH and S.-Y. SU 1285

Fig. 4. Postwall-filtered power as a function of acoustic pressure for a 2.25-MHz transducer with (a) 1-, (b) 3-, (c) 5-
and (d) 10-cycle transmitted pulses.

vivo being higher, the concentrations used in these de-
struction threshold experiments were higher than those
normally used clinically. The destruction threshold de-
pended on the concentration of the microbubbles, and
microbubbles at high concentrations tend to be much less
susceptible to destruction by ultrasound (Klibanov et al.
1998).
Signal processing
One M-mode image comprising 512 samples (i.e.,
3.2 mm) was acquired using 3,000 firing pulses (i.e., total
6 s) for each exposure setting and divided into 10 groups
of 300 pulses for statistical analysis. The acquired data
were first filtered with a second-order high-pass Butter-
worth filter with a cutoff frequency of 20 Hz over a slow
time index to remove the stationary echoes from the wall
of the tube, as well as wall reverberation echoes that
appeared directly below the tube. Figure 2a shows the
acquired RF signal, where the two dashed-line boxes
indicate the signals backscattered from the front and rear
walls of the tube. Figure 2b and c show the RF and
postwall-filtered envelop-detected M-mode images, re-
spectively. Note that envelop-detected image means to
filter out the carrier frequency component and to keep
only the magnitude of the remaining signal.
The postwall-filtered power, defined as the square
of the envelop-detected signal amplitude, was used to
assess the microbubble destruction threshold in our ex-
periments. The microbubbles were considered to be com-
pletely destroyed when the echo power received from the
imaging pulses was ⬍5% of that recorded before micro-
bubble destruction. The ideal received power would be
zero when the microbubbles were destroyed completely,
but there was always a low residual signal because of
background noise. Because microbubble echoes are de-
tected far above their linear resonance frequencies, and
the microbubbles are small compared with the wave-
length at 25 MHz, we assume that they behaved as
Rayleigh scatterers. Thus, the integrated backscattered
signal power was proportional to the number of intact
microbubbles within the tube. The postwall-filtered
power was first integrated over a time interval from
0 – 0.6 s, as shown in Fig. 2d, and the resulting curve was
integrated over depth. The integration was performed
over a region windowed in depth to reject duplicate
echoes because of reverberation. For consistency, the
same windowed region was used for all process.
To describe the degree of microbubble destruction,
we also considered the surviving percentage (SP) index,
which was defined as the ratio of integrated backscat-
tered power estimates with and without destruction pulse
insonation. Another parameter referred to destruction
1286 Ultrasound in Medicine and Biology
Fig. 5. Postwall-filtered power as a function of acoustic pressure for a 5-MHz transducer with (a) 1-, (b) 3-, (c) 5- and
(d) 10-cycle transmitted pulses.
percentage (DP) index was defined as 100% minus the
SP index. When the insonation destruction pulse was off
(i.e., no microbubble fragmentation), the estimated inte-
grated imaging power was normalized to 1 and the DP
index was 0%. All estimates of the integrated backscat-
tered power were normalized to this value.
EXPERIMENTAL RESULTS
Results for different insonation parameters
Figures 3 through 6 show the relationships between
DP values and acoustic excitation pressure (from 0 to 1
MPa) for 1-, 2.25-, 5- and 7.5-MHz pulses, respectively.
The hollow-circle symbols in Fig. 3a– d show the post-
wall-filtered normalized backscattered power integrated
over the pulse index (slow time in Fig. 2d) for destruc-
tion pulses comprising 1, 3, 5 and 10 cycles. The in-
sonation acoustic pressure was varied from 0 – 0.7 MPa
for each pulse length. The error bars in the figures show
one standard deviation estimated from 10 datasets re-
corded under each experimental condition.
Figure 3 shows that the pressure thresholds for
⬎90% UCA destruction were 0.5, 0.5, 0.4 and 0.1 MPa
for the 1-, 3-, 5- and 10-cycle pulses, respectively, indi-
cating that the pressure threshold for microbubble de-
struction decreases with increasing pulse length. Figures
4 – 6 have the same format as Fig. 3. The results show
Volume 34, Number 8, 2008
that the UCA destruction pressure threshold decreases
with decreasing pulse frequency and increasing pulse
length. More than 90% of the UCAs were destroyed at
0.1, 0.4, 0.6 and 0.7 MPa for 1-, 2.25-, 5- and 7.5-MHz
10 cycle–long destruction pulses, respectively.
Model for predicting the destruction pressure threshold
The results shown in Figs. 3– 6 indicate the presence
of an exponential relationship between the DP estimates
and insonation acoustic pressure. Therefore, we pro-
posed the following exponential model (i.e., f(x)) to fit
the discrete mean DP values weighted by the correspond-
ing reciprocal of standard deviations as a function of
pressure (i.e., weighted curve fitting):,
f(x) ⫽ (1 ⫺ aexp ⫺bx) ⫻ 100% (1)
where x is the pressure, a is the extrapolated DP value for
an acoustic pressure of zero (i.e., the f(x) intercept point
on the DP axis) and b is the intrinsic rate of decrease of
the exponential model. All nonlinear fits were performed
using a nonlinear least-squares algorithm (lsqnonlin
function in MATLAB). The fitting results are shown as
solid lines in Figs. 3– 6. The coefficient of determination
(R2) between the fitted curves and the original DP esti-
mates were all ⬎0.86. The high coefficients of determi-
nation indicate that our proposed model provides a good
 Microbubbles destruction ● C.-K. YEH and S.-Y. SU 1287

Fig. 6. Postwall-filtered power as a function of acoustic pressure for a 7.5-MHz transducer with (a) 1-, (b) 3-, (c) 5- and
(d) 10-cycle transmitted pulses.

description of the relationship between UCA destruction
and the acoustic pressure. In the following analyses, the
fitted model was interpolated to predict the degree of
UCA destruction under different insonation conditions.
The estimates of a and b obtained by fitting all the
experimental data shown in Figs. 3– 6 using eqn (1) are
listed in Table 2. From the results in Table 2, the b
estimates generally increase with increasing pulse length
and decrease with increasing the pulse frequency for
each identical pulse length condition. In other words, the
b estimate increasing is correlated with an increased
probability of UCA fragmentation.
Based on the model fitting results, the acoustic
pressure thresholds for DPs of 95%, 90%, 80% and 50%
for different pulse lengths and frequencies are shown in
Fig. 7a– d, respectively. Note that the error bars in these
bar charts are the 95% confidence intervals on the thresh-
olds taken from those fits at the indicated values of DP.
In Fig. 7a, the pressure threshold of a DP of 95% de-
creases with increasing pulse length for each pulse fre-
quency, and a lower pulse frequency results in a lower
destruction pressure threshold for each pulse length. In
Fig. 7a, the pressure thresholds for the pulse lengths of 3-
and 5-cycle in 2.25-MHz pulse is lower than that in

Table 2. The a, b and R2 estimates obtained from the exponential model with weighted curve fitting
Pulse Frequency Pulse Length 1-cycle 3-cycle 5-cycle 10-cycle

1 MHz a 0.967 0.967 0.955 0.998

b 5.379 5.931 6.622 21.759
R2 0.895 0.915 0.860 0.921
2.25 MHz a 0.955 0.958 0.973 0.985
b 4.603 6.120 7.632 10.129
R2 0.932 0.887 0.903 0.922
5 MHz a 1.019 1.015 1.026 0.989
b 2.734 3.220 3.480 3.875
R2 0.957 0.966 0.952 0.980
7.5 MHz a 1.019 0.984 1.008 1.077
b 2.329 2.599 3.002 2.939
R2 0.882 0.905 0.942 0.929
1288 Ultrasound in Medicine and Biology
Fig. 7. Microbubbles destruction pressure thresholds, with destruction percentage (DP) of (a) 95%, (b) 90%, (c) 80%
and (d) 50%.
1-MHz pulse. The possible reason is that the transient
response of the microbubble will be much faster in the
resonant case than off-resonance such that the radial
oscillation of the microbubble is expected to be large,
even for a short pulse (e.g., 3 to 5 cycles).
Degree of microbubble destruction vs. MI
The MI was derived by Apfel and Holland (1991) to
predict the likelihood of transient cavitation in diagnostic
ultrasound pressure fields. The formation and growth of
microbubbles occurs during the negative (i.e., rarefac-
tional) displacement of the pressure wave. The MI is
defined as the peak rarefactional pressure derated by 0.3
dB/cm/MHz (Pr(0.3)) divided by the square root of the
center frequency (fc) and thus is independent of both the
pulse length and PRF. Therefore, the MI predicts that
cavitation is less likely to occur at higher frequencies:
Pr(0.3)
MI ⫽ . (2)
兹fc
In this study, we used the measured value of peak
negative pressure to calculate the mechanical index
(MIe, where “e” indicates “experimental”). Figure 8a– d
shows the relationship between MIe estimates and DP
Volume 34, Number 8, 2008
values for pulse lengths of 1, 3, 5 and 10 cycles, respec-
tively. Figure 8 shows that, as the DP decreases, the MIe
estimate decreases for each combination of pulse fre-
quency and pulse duration. Further, the theoretical data
supporting the MI show that, at the threshold for inertial
cavitation, the ratio given in eqn (2) is nearly a single,
constant value. The results in Fig. 8a– d for different
pulse lengths do not support this prediction and thus the
results should not be interpreted as supporting the use of
the MI as the microbubbles destruction threshold. Be-
cause the MIe estimates should be independent of the
pulse frequency and corresponding pressure threshold
for an identical DP condition, the degree of UCA de-
struction is not related to the MI estimate.
DISCUSSION AND CONCLUSIONS
When microbubbles pass through a finite width
beam with a higher flow rate, the higher PRF of
destruction pulse number is required to ensure that the
bubbles within the sample volume were destroyed
completely. However, the higher PRF value of de-
struction pulse will induce the primary and secondary
radiation forces to affect the bubbles destruction.
Smith et al. (2007) demonstrated that the relationship
 Microbubbles destruction ● C.-K. YEH and S.-Y. SU 1289

Fig. 8. Experimental mechanical index (MIe) of microbubbles destruction thresholds with (a) 1-, (b) 3-, (c) 5- and (d)
10-cycle transmitted pulses.

between the PRF number of destruction pulse and
rapid fragmentation threshold of echogenic liposomes
is dependent. The lower-frequency pulses with higher
pulse number may lower a acoustic pressure threshold.
To further explore the degree of microbubble destruc-
tion with different PRF values of destruction pulse, we
did another similar experiment with PRF values from
250 Hz–16 kHz in 2.25-MHz destruction pulse. Note
that all experimental conditions were the same as
previous ones except the contrast agent was changed
to SonoVue microbubbles (Bracco Diagnostics, Inc.,
Milan, Italy). The results between DP estimates and
PRF are shown in Fig. 9. Figure 9a and b show the
results with acoustic pressures of 2.25-MHz destruc-
tion pulse of 0.2 and 0.4 MPa, respectively. The ‘Œ’
and ‘●’ symbols denote the pulse length of 1- and 3-cycle,
respectively. The results show that increasing pulse PRF
number increases the degree of microbubble destruction.

Fig. 9. Degree of microbubble destruction with different values of PRF from 250 Hz to 16 kHz under the conditions of
2.25-MHz destruction pulse with acoustic pressures of (a) 0.2 and (b) 0.4 MPa.
1290 Ultrasound in Medicine and Biology
In this study, we have used an acoustic-based ex-
perimental method to demonstrate the relationship be-
tween ultrasound exposure conditions and the degree of
microbubble destruction. Our proposed method repre-
sents a simple and easy way to determine the micro-
bubble destruction threshold in laboratory experiments.
The destruction threshold is determined based on a group
of microbubbles within a sample volume (and not just a
single bubble) and thus more closely replicates real clin-
ical conditions. The experimental results presented here
suggest that the acoustic excitation pressure, pulse center
frequency and pulse length affect the probability of mi-
crobubble fragmentation: increasing the pressure and the
pulse length and decreasing the pulse frequency are
correlated with an increased probability of fragmenta-
tion. These conclusions are consistent with those drawn
from optical observations (Bouakaz et al. 2005; Chomas
et al. 2000, 2001b). Note that fewer microbubbles were
observed to float up (because of buoyancy effect) and
accumulate at the top of the tube in such experiment
arrangement. To resolve the problem, the tube would be
held vertically to eliminate the retention of large micro-
bubbles at the top of the tube as a result of floatation.
We used an integrated backscattered power estimate
to represent the number of microbubbles that remain
within the sample volume after applying an insonation
destruction pulse. Theoretically, the intensity of the
backscattered signal is proportional to the relative
change in the microbubble concentration. Yamada et al.
(2005) found that the power of the received signal in
harmonic power Doppler imaging was proportional to
the bubble concentration for a constant applied acoustic
pressure. Schwarz et al. (1993) showed that the pulsed-
wave Doppler audio intensity is proportional to the con-
centration of Levovist over a limited range (i.e., for small
concentrations) and, hence, that the estimated backscat-
tered power is inversely proportional to the degree of
UCA destruction.
We have also used an exponential model with a
weighted curve fitting technique to estimate the DP as a
function of pressure. The high coefficients of determina-
tion (⬎ 0.86) for the fitting results in Figs. 3– 6 indicate
that the model provides a good description of UCA
destruction under different excitation pressures and,
hence, the model can be used to predict the pressure
threshold for UCA destruction.
Figure 8 indicates that there is no such relationship
between microbubble destruction and physical MI,
which is consistent with the conclusions that Smith et al.
(2007) drew from the microbubble destruction as de-
tected by changes in video intensity. Some limitations
such as background intensity and logarithmic compres-
sion will influence such video intensity measurements.
However, these do not apply to the present study, which
Volume 34, Number 8, 2008
used RF backscattered signals to estimate the degree of
microbubble destruction.
Predicting the degree of microbubble destruction is
useful in contrast-assisted perfusion imaging techniques.
Wilkening et al. (2000) proposed a flow estimation
model with fractional microbubble destruction based on
UCA time–intensity curves. The method is based on
continuous infusion of microbubbles resulting in a con-
stant UCA concentration in the blood. By a series of fast
high-energy destruction pulses, the UCA concentration is
decreased until a steady state is reached in which one
pulse destroys as much UCA as flows into the imaging
plane during one interpulse interval. Our proposed
acoustic-based experimental method can provide the in-
formation of degree of microbubbles destruction for
making such a flow estimation model feasible.
Another application is in high-resolution destruc-
tion/replenishment perfusion estimation systems. We
previously constructed a high-frequency UCA destruc-
tion/replenishment imaging system that exhibited a spa-
tial resolution of 160 ␮m in two dimensions (Yeh et al.
2004, 2007). That system used a 1-MHz focused trans-
ducer for microbubble destruction and a 25-MHz spher-
ically-focused transducer for pulse/echo imaging. Be-
cause the fractional UCA destruction at 25 MHz under a
specific pressure condition can be predicted using our
proposed method, our previous system could be simpli-
fied to use a single high-frequency transducer. Of course,
this would require modification to the theoretical de-
struction/replenishment model proposed by Wei et al.
(1998). Other potential applications include the induction
of microbubble cavitation and the use of sonoporation
for drug delivery and gene therapy.
Acknowledgements—We thank Mr. S.-Y. Lu for his assistance with the
experiments, and the reviewers for their valuable comments. This work
was supported by National Science Council under grant NSC 94-2320-
B-007-007.
REFERENCES
Apfel RE, Holland CK. Gauging the likelihood of cavitation from
short-pulse, low-duty cycle diagnostic ultrasound. Ultrasound Med
Biol 1991;17:179 –185.
Bouakaz A, Versluis M, de Jong N. High-speed optical observations of
contrast agent destruction. Ultrasound Med Biol 2005;31:391–399.
Bloch SH, Dayton PA, Ferrara KW. Targeted imaging using ultrasound
contrast agents. IEEE Eng Med Biol Mag 2004a;23:18 –29.
Bloch SH, Wan M, Dayton PA, Ferrara KW. Optical observation of
lipid- and polymer-shelled ultrasound microbubble contrast agents.
Appl Phys Lett 2004b;84:631– 633.
Chang PH, Shung KK, Wu SJ, Levene HB. Second harmonic imaging
and harmonic Doppler measurements with Albunex. IEEE Trans
Ultrason Ferroelectr Freq Control 1995;42:1020 –1027.
Chen WS, Matula TJ, Crum LA. The disappearance of ultrasound
contrast bubbles: Observations of bubble dissolution and cavitation
nucleation. Ultrasound Med Biol 2002;28:793– 803.
Chomas JE, Dayton P, May D, Allen J, Klibanov A, Ferrara KW.
Optical observation of contrast agent destruction. Appl Phys Lett
2000;77:1056 –1058.
 Microbubbles destruction ● C.-K. YEH and S.-Y. SU
Chomas JE, Dayton P, Allen J, Morgan K, Ferrera KW. Mechanisms of
contrast agent destruction. IEEE Trans Ultrason Ferroelectr Freq
Control 2001a;48:232–248.
Chomas JE, Dayton P, May D, Ferrara K. Threshold of fragmentation
for ultrasonic contrast agent. J Biomed Opt 2001b;6:141–150.
Chomas JE, Pollard R, Wisner E, Ferrara K. Subharmonic phase-
inversion for tumor perfusion estimation. Proc IEEE Ultrason Symp
2001c;2:1713–1716.
Dayton PA, Morgan KE, Klibanov AL, Brandenburger G, Nightingale
KR, Ferrara KW. A preliminary evaluation of the effects of primary
and secondary radiation forces on acoustic contrast agents. IEEE
Trans Ultrason Ferroelectr Freq Control 1997;44:1264 –1277.
Forsberg F, Liu JB, Merton DA, Rawool NM, Johnson DK, Goldberg
BB. Gray scale second harmonic imaging of acoustic emission
signals improves detection of liver tumors in rabbits. J Ultrasound
Med 2000;19:557–563.
Forsberg F, Shi WT, Merritt RB, Dai Q, Solcova M, Goldberg BB. On
the usefulness of the mechanical index displayed on clinical ultra-
sound scanners for predicting contrast microbubble destruction. J
Ultrasound Med 2005;24:443– 450.
Frinking P, Cespedes E, Kirkhorn J, Torp H, de Jong N. A new
ultrasound contrast imaging approach bases on the combination of
multiple imaging pulse and a separate release burst. IEEE Trans
Ultrason Ferroelectr Freq Control 2001;48:643– 651.
Goldberg B, Raichlen J, Forsberg F. Ultrasound Contrast Agents: Basic
Principles and Clinical Applications, ed 2. London: Martin Dunitz,
2001.
Hirokawa T, Nishikage T, Moroe T, Kajima M, Hayashi M, Naito T,
Yamane S, Shiota H. Visualization of uveal perfusion by contrast-
enhanced harmonic ultrasonography at a low mechanical index: A
pilot animal study. J Ultrasound Med 2002;21:299 –307.
Klibanov AL, Ferrara KW, Hughes MS, Wible JH, Wojdyla JK,
Dayton PA, Morgan KE, Brandenburger GH. Direct video-micro-
scopic observation of the dynamic effects of medical ultrasound on
ultrasound contrast microspheres. Invest Radiol 1998;33:863– 870.
Lotsberg O, Hovem JM, Askum B. Experimental observation of sub-
harmonic oscillations in Infoson bubbles. J Acoust Soc Am 1996;
99:1366 –1369.
O’Brien WD. Ultrasound– biophysics mechanisms. Prog Biophys Mol
Biol 2007;93:1–3:212–255.
Porter TM, Smith DAB, Holland CK. Acoustic techniques for assessing
the Optison destruction threshold. J Ultrasound Med 2006;25:1519 –
1529.
1291
Schwarz KQ, Bezante GP, Chen X, Schlief R. Quantitative echo
contrast concentration measurement by Doppler sonography. Ul-
trasound Med Biol 1993;19:289 –297.
Shi WT, Forsberg F, Hall AL, Chia RY, Liu JB, Miller S, Thomenius
KE, Wheatley MA, Goldberg BB. Subharmonic imaging with mi-
crobubble contrast agents: Initial results. Ultrason Imag 1999;21:
79 –94.
Shi WT, Forsberg F. Ultrasonic characterization of the nonlinear prop-
erties of contrast microbubbles. Ultrasound Med Biol 2000;26:93–
104.
Simpson DH, Chien TC, Burn PN. Pulse inversion Doppler: A new
method for detecting nonlinear echoes from microbubble contrast
agents. IEEE Trans Ultrason Ferroelectr Freq Control 1999;46:
372–382.
Smith DAB, Porter TM, Martinez J, Huang S, MacDonald RC,
McPherson DD, Holland CK. Destruction thresholds of echogenic
liposomes with clinical diagnostic ultrasound. Ultrasound Med Biol
2007;33:797– 809.
Wei K, Jayaweera AR, Firoozan S, Linka A, Skyba DM, Kaul S.
Quantification of myocardial blood flow with ultrasound-induced
destruction of microbubbles administered as a constant venous
infusion. Circulation 1998;97:473– 483.
Wei K, Le E, Bin JP, Coggins M, Thorpe J, Kaul S. Quantification of
renal blood flow with contrast-enhanced ultrasound. J Am Coll
Cardiol 2001;37:1135–1140.
Wilkening W, Postert T, Federlein J, Kono Y, Mattrey R, Ermert H.
Ultrasonic assessment of perfusion conditions in the brain and in
the liver. Proc IEEE Ultrason Symp 2000;2:1545–1548.
Yamada S, Komuro K, Mikami T, Kudo N, Onozuka H, Goto K, Fujii
S, Yamamoto K Kitabatake A. Novel quantitative assessment of
myocardial perfusion by harmonic power Doppler imaging during
myocardial contrast echocardiography. Heart 2005;91:183–188.
Yeh CK, Yang MJ, Li PC. Contrast-specific ultrasonic flow measure-
ments based on both input and output time intensities. Ultrasound
Med Biol 2003;29:671– 678.
Yeh CK, Ferrara W, Kruse DE. High-resolution functional vascular
assessment with ultrasound. IEEE Trans Med Imag 2004;23:1263–
1274.
Yeh CK, Lu SY, Chen YS. Microcirculation volumetric flow assess-
ment using high-resolution, contrast-assisted images. IEEE Trans
Ultrason Ferroelectr Freq Control 2008;55:74 – 83.