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Effects of droplet size and perfluorocarbon boiling point on the frequency

dependence of acoustic vaporization threshold

Article  in  The Journal of the Acoustical Society of America · February 2019

DOI: 10.1121/1.5091781

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Effects of droplet size and perfluorocarbon boiling point on the
frequency dependence of acoustic vaporization threshold
Mitra Aliabouzar, Krishna N Kumar, and Kausik Sarkara)
Department of Mechanical and Aerospace Engineering, The George Washington University,
Washington, DC 20052, USA

(Received 10 December 2018; revised 30 January 2019; accepted 4 February 2019; published
online 28 February 2019)
Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenic
microbubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are used
for therapeutic and diagnostic applications. This study systematically investigated the effect of
excitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds of
lipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane
(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smaller
than 3 lm and larger than 10 lm—in a tubeless setup. This study found that the ADV threshold
increases with frequency for the lowest boiling point liquid, PFP, for both large and small size
droplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization for
small size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The large
PFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFH
droplets, ADV threshold decreases with frequency that could possibly be due to the superharmonic
focusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-
olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholds
C 2019 Acoustical Society of America.
indicating that phase transition precedes inertial cavitation. V
https://doi.org/10.1121/1.5091781
[CCC] Pages: 1105–1116

I. INTRODUCTION cores and sizes, determining their ADV as well as inertial

cavitation (IC) thresholds, varying the excitation frequency.
Current commercially available microbubbles (MBs)
Since the pioneering study of Kripfgans et al. (2000), the
are limited to micrometer size range and have relatively
ADV of liquid PFC droplets has been investigated for many
short in vivo half-life due to premature dissolution (Katiyar
diagnostic and therapeutic applications (Fabiilli et al., 2010;
et al., 2009; Sarkar et al., 2009; Sheeran et al., 2013). The
Kopechek et al., 2013; Zhang et al., 2013; Vlaisavljevich
micrometer size distribution restricts them to the vascular
et al., 2015). The ADV threshold has been determined acous-
space and therefore they cannot be used for extravascular
tically as well as by direct optical observation of vaporization
interrogations (Ferrara et al., 2007). Production of MBs in
submicrometer range is challenging and would respond using high speed cameras. A review of the past studies using
poorly to ultrasound stimulation within the diagnostic range. both techniques has been provided in our previous paper
To mitigate these limitations, researchers are investigating (Aliabouzar et al., 2018). As noted there, ADV thresholds
clinical potentials of emulsions of phase shift micro- and obtained in different studies cannot be directly compared due
nanodroplets of volatile perfluorocarbon liquids (PFC) that to differences in the setups, droplet size distribution, and the
can be vaporized in situ into highly echogenic microbubbles method of threshold determination. However, some trends
by external application of ultrasound pulses (Kripfgans have been found to be consistent across experiments, e.g.,
et al., 2000; Fabiilli et al., 2009; Reznik et al., 2011; the ADV threshold decreases with increasing droplet size
Sheeran et al., 2013; Aliabouzar et al., 2018). The vaporiza- (Kripfgans et al., 2004; Fabiilli et al., 2009; Schad and
tion of the liquid core using acoustic waves, termed acoustic Hynynen, 2010; Sheeran et al., 2011), increasing temperature
droplet vaporization (ADV), occurs beyond a threshold peak (Zhang and Porter, 2010; Reznik et al., 2011), and increasing
negative pressure. We have recently determined the ADV number of cycles (Lo et al., 2007; Reznik et al., 2011).
threshold of an emulsion of perfluoropenetane (PFP) droplets However, ADV threshold showed an increasing trend with
as a function of frequency by investigating the scattered increasing excitation frequency when observed by optical
responses from the system (Aliabouzar et al., 2018). Here, means (Kripfgans et al., 2002; Kripfgans et al., 2004;
we extend the investigation to systems with different droplet Sheeran et al., 2013) and an opposite decreasing trend when
determined by echogenicity-based techniques (Kripfgans
et al., 2000; Schad and Hynynen, 2010; Williams et al.,
2013). In our previous article, we noted that the contradictory
a) results obtained in the literature have been ascribed by their
Present address: The George Washington University, 800 22nd Street
North West, Suite 3000, Washington, DC 20052, USA. Electronic mail: authors to limitations of the optical setup (Kripfgans et al.,
[email protected] 2004) as well as insufficient distinction between IC and

J. Acoust. Soc. Am. 145 (2), February 2019 0001-4966/2019/145(2)/1105/12/$30.00 C 2019 Acoustical Society of America
V 1105
ADV; experimentally measured IC threshold always The encapsulating shell contained lipids: 1,2-dipalmi-
increased with excitation frequency, the reason being the lon- toyl-sn-glycero-3-phosphatidylcholine (DPPC), 1,2-dipalmi-
ger continuous duration of negative pressure during the rare- toyl-sn-glycero-3-phosphate, sodium salt (DPPA), and 1,
factional half cycle (Apfel and Holland, 1991). We 2-dipalmitoyl-sn-glycero-3phosphoethanolamine-N-[methox-
acoustically determined the ADV threshold of a system of y(polyethylene glycol)-5000] (MPEG5000 DPPE). All the
PFP droplets in a tubeless setup using scattered (fundamental, lipids were purchased from Avanti Polar lipids (Alabaster,
sub-, and second harmonic) responses. We found the ADV AL). A total lipid concentration of 2 mg/ml was maintained
threshold for our system determined at room temperature to in the solution and the lipids were mixed in a molar ratio of
increase with increasing frequency (Aliabouzar et al., 2018). 11:82:7 (DPPA: DPPC: MPEG5000DPPE) in a solution of
The ADV threshold decreasing with increasing excitation fre- phosphate buffered saline (PBS), propylene glycol and glyc-
quency is nominally at odds with classical nucleation theory erol (8:1:1). These lipid components are similar to the ones
(CNT), and has been argued to be resulting from heteroge- used in Definity (Lantheus Medical Imaging, Inc., N.
neous nucleation or droplet deformation (Kripfgans et al., Billerica, MA, USA) microbubbles (Cheng, 2007). We fol-
2004) or nonlinear propagation and super-harmonic focus- lowed an established lipid solution preparation protocol used
ing (Li et al., 2014; Shpak et al., 2014; Miles et al., 2016). by Hui et al. (2017). Briefly, to prepare the lipid solution,
However, Shpak et al. (2014) noted that super-harmonic propylene glycol was heated to 10  C above the gel-to-liquid
focusing was only effective for larger droplets, as for transition temperature of the main phospholipid (DPPC) prior
smaller droplets one would need higher frequency of activa- to contacting with the lipid blend. When the propylene glycol
tion, where the attenuation of higher harmonics would also reached the desired temperature, lipids were added in the
be higher, effectively reducing the nonlinear distortion order of the least solubility—first DPPA, followed by DPPC,
phenomenon. and then MPEG5000 DPPE. Lipid solution was heated and
Here, we applied our acoustic methodology to droplets mechanically mixed for an hour (until the solution is clear).
of different average sizes and different liquid cores. Meanwhile PBS and glycerol, heated to the same temperature
Droplets of three different PFC liquid cores—perfluoropen- separately, were added to the lipid blend dropwise. The mix-
tane (PFP), perflurohexane (PFH), and perfluorooctyle bro- ture was later transferred into a preheated water bath (52  C)
mide (PFOB)—were investigated. The experiments were for another hour of bath sonication (35 kHz, VWR, West
performed at 37  C, above the bulk boiling point of PFP, but Chester, PA). The lipid solution was stored at 4  C and used
below those of the PFH and PFOB. For the highest bulk boil- within one week of preparation. Lipid-coated PFC droplets
ing point liquid, PFOB, experiments were also performed at were prepared according to an established protocol
43  C. The first objective of this study was to explore the (Kripfgans et al., 2000). Briefly, 2 ml of the lipid solution
effects of droplet size and the boiling point of PFC liquids on was added into a 3-ml glass vial (Wheaton industries Inc.,
the ADV and IC thresholds. The second objective was to Millville, NJ). The vial was sealed with a rubber stopper
investigate the frequency dependence of the ADV and IC (Wheaton Industries Inc.) and crimped. To remove the
thresholds of emulsions of different liquid cores (different trapped air in the headspace and minimize generation of air
boiling points and surface tensions) and droplet sizes. We bubbles, the vial was connected to a house vacuum pump for
used the scattered responses—peak-to-peak, fundamental, 30 s. Following that, 500 ll of PFC liquid was injected into
sub-, and second harmonic, as well as integrated power spec- the vial. Droplets of various PFC liquids were fabricated
trum in the low frequency range (500–900 kHz) for estima- using a Vial Mixer (Bristol Myers Squibb, North Billerica,
tion of ADV and IC thresholds, respectively. MA) shaken at 4800 rmp for 30 s. The vials were stored in
the fridge and were used on the same day of preparation.
II. EXPERIMENTAL METHODOLOGY Since PFCs are denser than water (Table I), larger drop-
lets would preferentially sediment to the bottom. To remove
A. Synthesis and characterization of droplets
extremely large droplets (>30 lm in diameter), we diluted
In this study, we have used three different PFC liquids: 2 ml of the droplet suspension (directly from the vial) with
PFP, PFH, and PFOB (99% wt. purity, FluoroMed, Round 13 ml degassed deionized (DI) water in a 15 ml centrifuge
Rock, TX, USA) as the liquid core. The properties of these tube (column length of 12 cm) and kept it in the fridge in a
PFC liquids are given in Table I (Cusco and Trusler, 1995; vertical position for a predetermined time. The predeter-
Hall et al., 2000; Strohm and Kolios, 2011). Note that the mined times were calculated according to Stokes equation: 5
surface tension listed is at the interface of the liquid and its min for PFP droplets, 4 min for PFH droplets, and 3 min for
vapor. PFOB droplets, allowing droplets larger than 30 lm in
TABLE I. Physical properties of PFC liquids used in this study.

Chemical Molecular Bulk boiling Surface Density Speed of sound Compressibility

PFC name Formula weight (g/mol) point (  C) tension (mN/m) (kg/m3) 25  C (m/s) 25  C (ms2/Kg)

Perfluoropentane (PFP) C5F12 288.03 28–30 9.5 1590 477 2.77  109
Perfluorohexane (PFH) C6F14 338.04 58–60 12.23 1648 548 2.02  109
Perfluorooctyle bromide (PFOB) C8BrF17 498.96 142  C 18 1930 630 1.3  109

1106 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.
FIG. 1. (Color online) Light microscopic images of diluted (a) larger than 30 lm, (b) between 10 and 25 lm, (c) smaller than 4 lm PFH droplets. Size distribu-
tion and concentrations of (d) large and (c) small groups of PFC droplets determined using qMicro and qNano systems. The scale bar is 50 lm.

diameter to settle (Feshitan et al., 2009). Following that, the 1 ml was removed to get rid of potential bubbles and the mid-
top 13 ml of the diluted droplet suspension was transferred dle 5 ml was transferred into a 50 ml tube to be used for the
into a new 15 ml centrifuge tube for the differential centrifu- ADV experiments.
gation process and the bottom 2 ml containing extremely The size distribution and the concentration of small droplet
TM
large droplets (>30 lm) was discarded. group were determined using a qNano (Izon Science ,
Droplets were then size isolated for diameters from 10 to Cambridge, MA). The droplet emulsion was diluted in PBS
25 lm (large droplet group) and smaller than 3 lm (small mixed with 0.03% Tween to ease passage of particles. For
droplet group) following a differential centrifugation technique these measurements, we used three different elastomeric
detailed in the literature (Feshitan et al., 2009; Mercado et al., Poly-urethane nanopores (NP) that covered a size range of
2016). To collect droplets of size from 10 to 25 lm, the droplet 200–1000 nm (NP800), 1000–4000 nm (NP2000), and
suspension was centrifuged at 50 g for 2 mins for PFP and 4000–10 000 nm (NP4000). Before and after each measure-
PFH droplets (40 g was used for PFOB since they are denser) ment, NPs were calibrated using appropriate plain polystyrene
using a bench type centrifuge (Eppendorf, NY). The top 3 ml particles (provided by Izon Company) of known size and num-
was discarded to get rid of potential bubbles formed during the ber. In our case, the applied stretching values were kept
mechanical agitation procedure. The bottom 2 ml containing between 45.5–47 mm. The voltage was set between 0.2 and
the target large droplets was transferred into a 50 ml tube to be 0.5 V to keep the current approximately 125 nA. For each sam-
used for ADV experiments. To isolate droplets smaller than ple, 1000 particles were counted and analyzed using the qNano
3 lm, the middle 8 ml of the suspension was transferred into a software. For the large droplet group, qMicro with micropore-
new tube for the second cycle of centrifugation at 60 g for membrane MP25 (size range of 5–25 lm) was used. For larger
1 min for PFP, PFH, and PFOB droplets. Similarly, the top droplets, we also performed light microscopy image analysis

TABLE II. Average diameter of the droplets using qNano and qMicro measurements.

Small droplets Large droplets

Liquid core Diameter (nm) Concentration (particles/ml) Diameter (lm) Concentration (particles/ml)

PFP 947 6 45 6.1 109 11.99 6 3.7 8.6  104

PFH 860 6 100 3.4 109 14.21 6 3.8 1.8  105
PFOB 954 6 134 2.36 109 15.44 6 3.4 1.18  105

J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al. 1107
 FIG. 2. (Color online) (a) Schematic
representation (horizontal plane show-
ing the transducers confocally aligned
is shown in top view). (b) Picture of
the experimental setup used for deter-
mining ADV threshold. The droplet
emulsion flows from an immersed
metal tube [see (b)] 5 mm above the
outer diameter of the transducers.

techniques which were in good agreement with qMicro results receiving transducers were confocally positioned at right
(data not shown). Figures 1(a)–1(c) show light microscope angles (Fig. 2). All the transducers were calibrated in a free
(AmScope FMA050, MA at 10) images of isolated and field using a calibrated capsule hydrophone (HGL0200,
diluted PFH droplets of size 30 lm, 10–25 lm, and smaller dynamic range 1–20 MHz, Onda, Sunnyvale, CA).
than 3 lm, respectively. Size measurement results with qMicro For calibration, the transducer and the hydrophone were
and qNano are shown in Figs. 1(d) and 1(e), respectively. submerged in a water tank filled with degassed DI water.
Weighted average diameters of both small and large size drop- The pressure amplitudes at the focus of the transducers were
lets of different PFC liquids are summarized in Table II. measured and recorded with the capsule hydrophone at low
Droplet suspension was diluted 200 X and 50 X for driving voltages—from 0.15 to 0.6 MPa peak negative pres-
small and large suspensions, respectively, prior to the ADV sures. For higher peak negative pressures [from 0.6 to
threshold determination experiments. The droplet suspension 4 MPa; Mechanical Index (MI) for the highest value is 2.6],
was then checked optically (microscope) and acoustically to avoid hydrophone damage due to cavitation, nonlinear
(see Sec. II B below) for any bubble presence. numerical modeling based on the Khokhlov-Zabolotskaya-
Kuznetsov (KZK) equation was used. We have verified this
B. Experimental setup for ADV and IC thresholds calibration method by plotting the axial pressure distribution
determination of a 2.25 MHz transducer (from the transducer surface up to
a distance of 35 mm) at 450 kPa using the hydrophone and
For ADV experiments, typically the droplet suspension is
compared with the KZK simulations. The results were in
passed through the focal volume of a transducer in a flow good agreement (data not shown). This calibration technique
setup. In our previous work, we have shown that the focal vol- has also been verified by Bessonova and Wilkens (2013) and
ume is small (4.21 mm3 at 2.25 MHz) and it decreases with Canney et al. (2008) using both fiber-optic hydrophone
increasing frequency (Aliabouzar et al., 2018). Similar to that measurements and KZK predictions.
work, here we have also used a tube-less setup to avoid wall For ADV experiments, an arbitrary/function generator
effects (e.g., beam-diffraction, standing waves, reflection, and (Agilent, 33250A, Santa Clara, CA) was utilized to generate an
scattering from the wall) and any potential complex acoustic 8-cycle sinusoidal pulse at a pulse repetition frequency (PRF)
field at the focus. Figures 2(a) and 2(b) show a representation of 100 Hz. This signal was then amplified using a 55 dB power
of our experimental setup for ADV threshold determination. A amplifier (A-150, ENI, Rochester, NY) and used to excite the
chamber (80  60  60 mm) made of polycarbonate was filled transmitting transducer. The scattered signal was passively
with degassed DI water. To perform ADV studies, we received by the receiving broadband transducer connected to
employed three spherically focused immersion transducers a pulser/receiver (5800, Panametrics-NDT, Waltham, MA) in
(Panametrics Transducer, Olympus NDT Corporation, a receiving mode with a 20 dB gain (HP filter: 100 kHz, LP
Waltham, MA), each having an element diameter of 1.27 cm filter 35 MHz). The amplified signals were then fed into
and a focal length of 3.05 cm as transmitters (f-number ¼ 2.4). the oscilloscope (Tektronix, MDO3024, Beaverton, OR) to
Transducers with central frequencies of 2.25 MHz (Model view them in real time. Signals were acquired directly from
V306; 6 dB: 1.46–3.2 MHz), 10 MHz (Model V311; 6 dB: the oscilloscope using MATLAB (Mathworks, Natick, MA).
6.96–13.16 MHz), and 15 MHz (Model V319; 6 dB: Fifty voltage-time radio frequency (RF) traces were acquired
10.10–18.97 MHz) were used as transmitters at their central in a sample mode and stored for further processing.
frequencies. A broadband, cylindrically focused transducer Measurement for each setting was repeated five times.
(Sonic Concepts, Bothell, WA, USA) with an active diameter To acoustically confirm that the freshly prepared droplet
of 17.5 and 50 mm geometric focus was used as the suspension was bubble-free, 20 ll of suspension was dis-
receiver. This transducer had a flat frequency response persed in the solution chamber and then was excited by ultra-
between 10 kHz and 20 MHz. The transmitting and sound at an excitation pressure of 150 kPa, which was below

1108 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.
FIG. 3. (Color online) (a) RF trace of control and large PFP droplet signal at 0.35 MPa (below vaporization), 0.95 MPa (above vaporization without IC), and
3 MPa (above vaporization with IC); (b) corresponding fast Fourier transform from the control and the PFP droplet suspension at 2.25 MHz.

the minimum ADV thresholds found in this study. The cross-section of a bubble is several orders of magnitude
scattered response was found to be comparable with that of higher than that of a liquid droplet of the same size (Medwin
the background noise confirming the suspension to be bubble and Clay, 1997). The acoustic behavior of gas bubbles is
free. Note that the current setup was found capable of detect- mainly dominated by the resonance. Around the resonant
ing acoustical signals from even a very small amount (4 ll) frequency the effects of backscattering and absorption are
of a microbubble suspension dispersed in the same chamber. much enhanced (Sarkar and Prosperetti, 1994). Furthermore,
A syringe pump (KD Scientific Inc, Holliston, MA) was used upon vaporization, the droplet undergoes volumetric expan-
to inject the droplet suspension into the solution chamber with sion of about 125 fold, as shown by Rapoport et al. (2009)
a flow rate and particle velocity of 20 ll/s and 2 mm/s, respec- and Sheeran et al. (2011), resulting in a sudden jump in their
tively. The syringe exit was connected to a stainless steel tube scattered response. Here, we plotted peak-to-peak, funda-
(inner diameter 2.9 mm). The tube exit was immersed in the mental, and non-linear components, such as sub or second
solution chamber and mounted approximately 5 mm above harmonics (when present) as the ultrasound excitation ampli-
the outer diameter of the transducers. The distance was suffi- tude was progressively increased.
cient so that the steel tube would not generate any signal. The For IC studies, experiments were done in the same
steel tube was attached to the manual linear translation stage setup. Inertial cavitation is accompanied by a broadband
(Newport Corporation, Irvine, CA) having a precision of noise. The IC threshold here was defined as the amplitude at
1 lm in all three directions. The dimensions of the focal which the integrated power calculated in the low frequency
region for a given frequency has been calculated by the for- range, from 500 to 900 kHz, increased from the baseline.
mulae from the transducer manual (Olympus, 2006). We Note that the low frequency emission has been used as IC
ensured that the stream of droplets passed through the inter- criterion in several past studies (Giesecke and Hynynen,
secting focal volume by first injecting a stream of propylene 2003; Fabiilli et al., 2009; Moncion et al., 2016). Figure 3
glycol (higher acoustic impedance of 1.61 MRayl compared represents raw RF data along with the corresponding fre-
to that of water 1.48 MRayl) into the solution chamber via the quency spectrum for the control (base lipid solution) and the
steel tube to determine the proper position of the tube. When droplet signals at three different excitation amplitudes of
the steel tube was properly mounted so that the stream of pro- 0.35 MPa (below ADV), 0.95 MPa (above ADV/without IC),
pylene glycol passed through the focal volume, we obtained a and 3 MPa (above ADV with IC) at 2.25 MHz for large PFP
strong signal. The positioning procedure was repeated before droplets. The droplet signal at 0.35 MPa was very close to
each experiment. All experiments were performed at 37  C. the control. For an excitation pressure 0.95 MPa, above the
PFOB droplets, due to higher bulk boiling point of 142  C, ADV threshold, the droplet signal became substantially
did not experience ADV at 37  C; the experiments for PFOB higher than the control. This sharp increase can also be noted
droplets were repeated at 43  C. Degassed DI water and the in the corresponding frequency spectrum. We found that the
droplet suspensions were kept in water bath heated to the peak-to-peak voltage, fundamental and nonlinear compo-
desired temperature and allowed to reach equilibrium. To nents (if present) all increased from the baseline readings
determine the ADV threshold, the excitation pressure was and continued to be high for all the subsequent amplitudes.
increased in steps of 200 kPa for each excitation frequency For an objective definition of the threshold values, data were
from 100 kPa to 4 MPa. segmented between, before, and after a sudden increase and
then a piece-wise linear fit was performed on each segment
C. Criterion for deciding the acoustic droplet using OriginLab (OriginPro8, OriginLab, Northampton,
vaporization and inertial cavitation thresholds
MA). The ADV as well as IC thresholds were defined as the
Liquid droplets are poor scatterers of ultrasound while peak negative pressure corresponding to the intersection of
microbubbles are far stronger scatterers. The scattering the first two segmental lines of the piece-wise linear fit. Such

J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al. 1109
methods of threshold determination have been used in sev- The signal from the droplet suspension was comparable
eral past studies (Radhakrishnan et al., 2013; Radhakrishnan to the control signal at excitation amplitudes below 0.4 MPa
et al., 2016; Moncion et al., 2017). In view of the stochastic for PFP droplets and 2.3 MPa for PFH droplets in the plots.
nature, the ADV threshold values, although determined here In Fig. 4(a), the intersection of the first two segments of the
as in the previous investigations through a quantitative piece-wise linear fit (ADV threshold) happens at 0.4 MPa
method, are to be understood as a representative of the val- (R2 ¼ 0.99) and 2.3 MPa (R2 ¼ 0.98) for PFP and PFH drop-
ues of excitation where vaporization occurs, rather than a lets, respectively. This was using the peak-to-peak data. The
precise single value, with no vaporization below that range. fundamental component shown in Fig. 4(b) resulted in very
similar values of the threshold—0.39 MPa (R2 ¼ 0.99) and
D. Statistical analysis 2.27 MPa (R2 ¼ 0.98) for PFP and PFH droplets, respec-
tively. Note that the experiments were performed at 37  C.
All the scattered responses were presented as the mean of We did not observe any vaporization for PFOB droplets at
the data sets acquired at each setting and their corresponding 37  C (data not shown) for the excitation pressures (up to
standard deviations. All experiments at each driving pressure 4 MPa) studied here. Even at 43  C, PFOB droplets failed to
and frequency were repeated five times. A Kolmogorov- vaporize [response is same as control in Figs. 4(a) and 4(b)].
Smirnov test was performed on the data to ensure a normal Note that the scattered fundamental response from PFP drop-
distribution. Finally, the ADV threshold values from different lets post ADV seemed to saturate at higher amplitude, a phe-
criteria (scattered responses) were averaged. nomenon also seen previously in ADV studies of PFP
droplets (Reznik et al., 2011). Similar saturation was also
III. RESULTS AND DISCUSSION observed in experimental measurement as well as numerical
prediction of scattered responses from coated microbubbles
A. Acoustic droplet vaporization at 2.25 MHz
(Sarkar et al., 2005; Paul et al., 2010; Paul et al., 2013; Paul
Figures 4(a), 4(b), 4(c), and 4(d) plot the peak-to-peak et al., 2014).
voltage, fundamental, sub, and second harmonic responses, Using a similar procedure on the sub- and second-
respectively, from the droplet suspensions of PFP, PFH, and harmonic components [Figs. 4(c) and 4(d)] for the large
PFOB from the large size distribution group at the excitation droplet suspension, we obtained the same threshold values of
frequency of 2.25 MHz. 0.41 [(R2 ¼ 0.93) and 0.36 (R2 ¼ 0.98) for PFP droplets as in

FIG. 4. (Color online) (a) Peak-to-peak voltage, (b) fundamental, (c) subharmonic, and (d) second harmonic components of the scattered response for large
droplets with PFP, PFH, and PFOB (at 43  C) liquid cores at the excitation frequency of 2.25 MHz.

1110 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.
FIG. 5. (Color online) (a) Peak-to-peak, (b) fundamental, (c) sub, and (d) second harmonic components for suspensions of small droplets with various PFC
cores at 2.25 MHz.

Figs. 4(a) and 4(b)]. However, PFH droplets did not generate above 2.9 MPa. However, at this high excitation, second har-
any subharmonic response. Note that PFH droplets, with monic response can also be due to nonlinear propagation of
boiling point higher than the operating temperature, have ultrasound in water (Clay and Medwin, 1977) rather than from
been observed by optical means to undergo repeated vapori- bubble activities. Specifically, for PFOB droplets at 43  C, we
zation and re-condensation in response to irradiation by see an increase in second harmonic response, although, as
near-infrared laser pulses (Hannah et al., 2016). B-mode already noted above, these droplets did not register any
observations of similar phenomenon (Asami and Kawabata, increase in other scattered components. In our previous study,
2012) also reported a transient vaporization of PFH droplets we investigated the contribution of nonlinear propagation in
compared to PFP droplets. The transient nature of the vapori- water by recording the second harmonic response from a non-
zation or re-condensation and therefore not enough time for vaporizing liquid propylene glycol stream. The effect of non-
sustained oscillation could be the reason for the lack of subhar- linear propagation was not pronounced for lower amplitudes
monic response. PFH is a stronger liquid (i.e., one with higher (below 2.5 MPa) but it slowly built up at higher amplitudes at
boiling point and surface tension, see Table I) compared to this excitation frequency (Aliabouzar et al., 2018).
PFP, and therefore requires substantially higher amplitude In our previous study (Aliabouzar et al., 2018), analyz-
(2.3 MPa peak negative pressure compared to 0.4 MPa peak ing scattered responses at different frequency components—
negative pressure for PFP at this frequency) for vaporization. fundamental, sub-, and second-harmonic—resulted in simi-
At 2.3 MPa peak rarefactional pressure, the peak compres- lar but slightly different ADV thresholds. There, we mea-
sional pressure is also much higher (4.25 MPa), which might sured different scattered components performing separate
be another reason to favor re-condensation of PFH droplets. experiments using different narrowband transducers as
Note that, at the ambient temperature of 37  C, PFP is super- receiver. In contrast, here we used a broadband transducer
heated according to their bulk boing point while PFH is not. and the different frequency components gave rise to virtually
Incidentally, repeated vaporization and re-condensation offer same ADV threshold values for both sizes and all excitation
potential applications where extended imaging or drug deliv- frequencies (see also below).
ery is required (Asami and Kawabata, 2012). Figure 4(d) dis- In Fig. 5, we plot peak-to-peak, fundamental, sub-,
plays the second harmonic generation at 2.25 MHz. Similar to and second-harmonic responses for the small droplets of
other scattered responses, second harmonic also rose at PFP and PFH droplets at 2.25 MHz. Similar to the large
0.36 MPa for PFP. For PFH droplets, this jump was noticeable droplets the procedure applied to the peak-to-peak,

J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al. 1111
FIG. 6. (Color online) (a) Peak-to-peak and (b) fundamental responses of suspensions of large droplets, (c) peak-to-peak and (d) fundamental responses from
small droplets at the excitation frequency of 10 MHz.

fundamental, sub-, and second-harmonic data resulted in B. Acoustic droplet vaporization at 10 MHz
very similar vales of threshold for PFP small droplets:
Figures 6(a) and 6(b) plot the peak-to-peak voltage and
0.52 MPa (R2 ¼ 0.97), 0.56 MPa (R2 ¼ 0.99), 0.5 MPa
the fundamental response from large PFP and PFH droplet
(R2 ¼ 0.99), and 0.49 MPa (R2 ¼ 0.99), respectively. No
groups as functions of peak negative pressure at 10 MHz
vaporization was observed for small PFH and PFOB (data
excitation frequency. At this frequency, thresholds of vapori-
not shown) droplets (up to 4 MPa). For PFOB, we also
zation were 0.58 (R2 ¼ 0.99) [Fig. 6(a)] and 0.56 (R2 ¼ 0.92)
repeated the experiments at 43  C and did not record any
[Fig. 6(b)] for large PFP and 1.52 (R2 ¼ 0.91) [Fig. 6(a)] and
vaporization (data not shown).
1.58 MPa (R2 ¼ 0.97) [Fig. 6(b)] for large PFH droplets. For
Comparing Figs. 4 and 5, one can notice the effect of
small PFP droplets, ADV threshold was recorded as
droplet size on average ADV threshold for each droplet sus-
1.58 MPa (R2 ¼ 0.93) and 1.66 MPa (R2 ¼ 0.99) in Figs. 6(c)
pension at 2.25 MHz. The ADV threshold increased from
and 6(d), respectively. We did not detect any vaporization
0.39 MPa for large PFP to 0.52 MPa for small PFP droplets.
for small PFH droplet group at this excitation frequency (up
For PFH, while large droplets recorded 2.28 MPa, no vapori-
to 4 MPa) similar to 2.25 MHz. Similarly, no vaporization of
zation was observed for small droplets.
either small or large PFOB droplets (even at 43  C) was
The threshold of vaporization for small PFP droplets at
detected at this frequency either (data not shown). At the
37  C (0.52 MPa) was lower compared to the one we reported
excitation frequency of 10 MHz, we did not detect subhar-
in the previous study at room temperature (1.05 MPa) as can
monic response from the droplet suspensions at the vaporiza-
be expected (Zhang and Porter, 2010; Reznik et al., 2011).
tion threshold, as was also the case in our previous paper,
Higher boiling point PFC liquids such as PFH and PFOB
possibly due to the fact that subharmonic response from a
have not been explored as much due to the low efficiency of
bubble is frequency and threshold dependent (Katiyar and
vaporization. Zhang and Porter (2010) observed no vaporiza-
Sarkar, 2011).
tion for H-PFP (2H, 3H-perfluoropentane; boiling point of
56  C) droplets having an average diameter of 690 6 233 lm
C. Acoustic droplet vaporization at 15 MHz
at the excitation frequency of 2 MHz up to pressure values as
high as 8 MPa. Moncion et al. (2017) reported a threshold At the excitation frequency of 15 MHz, Fig. 7(a) recorded
value of 2.2 MPa for double emulsions of size 13.9 6 0.04 lm ADV vaporization thresholds of 1.09 MPa (R2 ¼ 0.99),
made of PFH liquid at 2.5 MHz. 1.12 MPa (R2 ¼ 0.98) and 2.8 MPa (R2 ¼ 0.87) for large PFP,

1112 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.
FIG. 7. (Color online) (a) Peak-to-peak, (b) fundamental response from large PFC droplets, (c) peak-to-peak, and (d) fundamental response from small PFC
droplets at the excitation frequency of 15 MHz.

PFH, and PFOB (only at 43  C) droplets using the peak-to- of ADV threshold on the frequency of excitation. We have
peak response. Corresponding values from fundamental recorded an increasing trend of vaporization threshold with
response were again very similar—1.06 MPa (R2 ¼ 0.97), frequency for both small and large PFP (the most volatile
1.1 MPa (R2 ¼ 0.98) and 2.77 MPa (R2 ¼ 0.91), respectively liquid considered here with the bulk boiling point of 29  C
[Fig. 7(b)]. The small droplets expectedly showed a higher and surface tension of 9.5 mN/m) droplets. The increasing
value of ADV threshold—2.57 MPa (R2 ¼ 0.96) for PFP drop- trend of the ADV threshold with increasing frequency can be
let [Fig. 7(c)]. This one was from the peak-to-peak response, reasoned as arising from longer duration of continuous nega-
which was again very similar to the one obtained from funda- tive pressure that the droplets experience at lower frequency,
mental is 2.55 MPa (R2 ¼ 0.93) [Fig. 7(d)]. No vaporization which in turn would increase the probability of nucleation.
was detected for small PFH and PFOB droplets. According to homogenous nucleation theory, liquid under
Matsuura et al. (2009) reported an ADV threshold of negative pressure is metastable and given sufficient time
4.7 MPa for PFH droplets at the excitation frequency of 18 vapor bubbles nucleate and expand (Church, 2002). As noted
MHz in a cellulose tube positioned at the focus of the linear before, ADV increasing with frequency of excitation has
array. Note that direct comparison cannot be made due to the been reported in our previous paper for PFP droplets at
differences in droplet sizes, and experimental setup. 20  C as well as by others (Kripfgans et al., 2004; Martin
However, the higher value in their study might partially be et al., 2012; Sheeran et al., 2013).
due to increased attenuation across the wall of the cellulose However, here we also observed an opposite trend for a
tube. higher boiling point PFC liquid, PFH—ADV threshold
decreases with frequency of excitation for large droplet
group as shown in Fig. 8. PFOB large droplets did not vapor-
D. Frequency dependence of ADV threshold on size
ize at lower frequencies in the range of excitations (<4 MPa)
and boiling point of PFCs
considered here, but registered a threshold of 2.8 MPa at
As noted in the Introduction, previous studies of the fre- 15 MHz (also at 43  C). Note that PFH and PFOB are stron-
quency dependence of ADV thresholds of PFC liquid drop- ger liquids, i.e., with higher boiling points and surface ten-
lets have been contradictory. Here, we have combined the sions than PFP. They require much higher peak rarefactional
effects of size and liquid core properties on the dependence pressures for vaporization where we expect to have

J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al. 1113
 excitation frequency depending on size and droplet core. It
shows that both size and material properties of the liquid
core, such as boiling point and surface tension, play a critical
role in defining the trend of vaporization with frequency of
excitation. The average thresholds for the emulsions of dif-
ferent sizes and liquid cores are summarized in Fig. 8. Note
that the MI calculated for this system ranged from 0.26
(0.39 MPa at 2.25 MHz for large PFP droplets) to 1.51
(2.27 MPa at 2.25 MHz for large PFH droplets).

E. Inertial cavitation
For the IC threshold measurements, we plot the inte-
grated power spectrum within 500–900 kHz frequency inter-
val (Fig. 9). Similar to ADV threshold detection, piece-wise
FIG. 8. (Color online) ADV thresholds of PFC droplets as a function of fre- linear fitting was performed on the data points to find the IC
quency of excitation for PFP small and large (37  C), PFH large (37  C), and threshold. We have plotted the integrated power-frequency
PFOB large (43  C) groups. spectrum for both small and large groups of PFP droplets at
2.25 MHz [Fig. 9(a)], 10 MHz [Fig. 9(b)], and 15 MHz [Fig.
significant amount of nonlinearity. Therefore, superharmonic 9(c)]. IC thresholds are as follows: at 2.25 MHz, 1.6 MPa
focusing possibly took place at these high amplitudes for (R2 ¼ 0.96) for small PFP and 1.12 MPa (R2 ¼ 0.91) for large
large PFH and PHOB droplets. Superharmonic focusing has PFP droplets; at 10 MHz, 2.77 MPa (R2 ¼ 0.86) for small
been optically shown to be pronounced for larger droplets PFP and 2.09 MPa (R2 ¼ 0.84) for large PFP droplets; at
(radius >4 lm), higher frequencies and higher amplitudes 15 MHz: 3.47 MPa (R2 ¼ 0.7) for small PFP and 2.57 MPa
(Shpak et al., 2014). It is noteworthy that we see both (R2 ¼ 0.86) for large PFP droplets. At each frequency, as
trends—ADV threshold increasing and decreasing with expected, IC threshold for larger droplets are higher than

FIG. 9. (Color online) IC threshold at (a) 2.25 MHz, (b) 10 MHz and (c) 15 MHz for small as well as large PFP droplets, and (d) comparison of IC and ADV
for small and large PFP droplets at various frequencies.

1114 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.
that of the smaller droplets, and IC threshold is always Bessonova, O. V., and Wilkens, V. (2013). “Membrane hydrophone mea-
higher than that for ADV [Fig. 9(d)]. For PFH and PFOB surement and numerical simulation of HIFU fields up to developed shock
regimes,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control 60, 290–300.
cases, no IC was recorded for the pressures studied here. Canney, M. S., Bailey, M. R., Crum, L. A., Khokhlova, V. A., and
Moncion et al. (2017) reported an IC threshold value of Sapozhnikov, O. A. (2008). “Acoustic characterization of high intensity
4.8 MPa for double emulsions of PFH droplets of similar focused ultrasound fields: A combined measurement and modeling
approach,” J. Acoust. Soc. Am. 124, 2406–2420.
size (14 lm) at 2.5 MHz within a fibrin gel. An IC threshold
Cheng, K. T. (2007). “Perflutren lipid microspheres,” Molecular Imaging
of 1.5 MPa was reported for both PFP and PFH droplets of and Contrast Agent Database (MICAD), Bethesda, MD.
2 lm in diameter by Giesecke and Hynynen (2003) at Church, C. C. (2002). “Spontaneous homogeneous nucleation, inertial cavi-
2.18 MHz. tation and the safety of diagnostic ultrasound,” Ultrasound Med. Biol. 28,
1349–1364.
Clay, C. S., and Medwin, H. (1977). Acoustical Oceanography: Principles
IV. CONCLUSION and Applications (Wiley, New York).
Cusco, L., and Trusler, J. (1995). “Identification of environmentally accept-
We systematically studied the effect of frequency of able low-sound speed liquids,” Int. J. Thermophys. 16, 675–685.
excitation on the threshold of vaporization by performing Fabiilli, M. L., Haworth, K. J., Fakhri, N. H., Kripfgans, O. D., Carson, P.
ADV experiments on lipid coated droplets of different sizes L., and Fowlkes, J. B. (2009). “The role of inertial cavitation in acoustic
and liquid cores. Progressively increasing the peak negative droplet vaporization,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control 56,
1006–1017.
excitation amplitude, we found a simultaneous sharp change Fabiilli, M. L., Haworth, K. J., Sebastian, I. E., Kripfgans, O. D., Carson, P.
in peak-to-peak, fundamental, sub- and second harmonic L., and Fowlkes, J. B. (2010). “Delivery of chlorambucil using an
scattered responses at a certain value of the excitation, indi- acoustically-triggered perfluoropentane emulsion,” Ultrasound Med. Biol.
cating vaporization. The ADV threshold, where the sharp 36, 1364–1375.
Ferrara, K., Pollard, R., and Borden, M. (2007). “Ultrasound microbubble
change in the scattered responses occurs, was found to contrast agents: Fundamentals and application to gene and drug delivery,”
increase with frequency for both small and large size PFP Ann. Rev. Biomed. Eng. 9, 415–447.
droplets (the most volatile PFC liquid studied here). For Feshitan, J. A., Chen, C. C., Kwan, J. J., and Borden, M. A. (2009).
large PFH droplets—small ones did not vaporize in the exci- “Microbubble size isolation by differential centrifugation,” J. Colloid
Interf. Sci. 329, 316–324.
tation range (up to 4 MPa) considered—the ADV threshold Giesecke, T., and Hynynen, K. (2003). “Ultrasound-mediated cavitation
decreased with frequency. The large PFOB droplets vapor- thresholds of liquid perfluorocarbon droplets in vitro,” Ultrasound Med.
ized only at the highest frequency (15 MHz), indicating the Biol. 29, 1359–1365.
Hall, C. S., Lanza, G. M., Rose, J. H., Kaufmann, R. J., Fuhrhop, R. W.,
same trend. Note that, while the increasing trend of ADV
Handley, S. H., Waters, K. R., Miller, J. G., and Wickline, S. A. (2000).
threshold with frequency is due to increased probability of “Experimental determination of phase velocity of perfluorocarbons:
nucleation, the decreasing trend for larger droplets and Applications to targeted contrast agents,” IEEE Trans. Ultrason.
higher boiling point liquids may be due to superharmonic Ferroelectr. Freq. Control 47, 75–84.
Hannah, A. S., Luke, G. P., and Emelianov, S. Y. (2016). “Blinking phase-
focusing facilitated by the nonlinearity at the higher excita- change nanocapsules enable background-free ultrasound imaging,”
tion in those cases as well as larger droplet radii. These find- Theranostics 6, 1866–1876.
ings show that both size and physical properties of the liquid Hui, P. K., Bishop, J. E., and Madrigal, E. S., Jr., (2017). “Preparation of a
core such as boiling point and surface tension play a critical lipid blend and a phospholipid suspension containing the lipid blend,”
U.S. patent no. 20010003580A1.
role in defining the trend of vaporization with frequency of Katiyar, A., and Sarkar, K. (2011). “Excitation threshold for subharmonic
excitation. IC was measured by calculating the integrated generation from contrast microbubbles,” J. Acoust. Soc. Am. 130,
power frequency spectrum in the range from 500 to 900 kHz. 3137–3147.
IC was observed only for PFP droplets (under peak negative Katiyar, A., Sarkar, K., and Jain, P. (2009). “Effects of encapsulation elas-
ticity on the stability of an encapsulated microbubble,” J. Colloid Interf.
pressures up to 4 MPa) and it increased with the frequency Sci. 336, 519–525.
of excitation and was lower for larger droplet sizes. For all Kopechek, J. A., Park, E., Mei, C.-S., McDannold, N. J., and Porter, T. M.
the frequencies studied here, ADV thresholds were lower (2013). “Accumulation of phase-shift nanoemulsions to enhance MR-
than the IC thresholds. guided ultrasound-mediated tumor ablation in vivo,” J. Healthcare Eng. 4,
109–126.
Kripfgans, O. D., Carson, P. L., and Fowlkes, J. B. (2002). “On the mecha-
ACKNOWLEDGMENTS nism of acoustic droplet vaporization,” in Proceedings of the 2002 IEEE
Ultrasonics Symposium, October 8–11, Munich, Germany, pp. 535–538.
We would like to thank Dr. A. Pal and A. Senthikumar Kripfgans, O. D., Fabiilli, M. L., Carson, P. L., and Fowlkes, J. B. (2004).
from Izon Science Ltd for performing the qMicro size “On the acoustic vaporization of micrometer-sized droplets,” J. Acoust.
measurements. K.S. acknowledges partial support from NSF Soc. Am. 116, 272–281.
Kripfgans, O. D., Fowlkes, J. B., Miller, D. L., Eldevik, O. P., and Carson,
Grant Nos. CBET-1205322, CBET 1603639, and NIH
P. L. (2000). “Acoustic droplet vaporization for therapeutic and diagnostic
R01GM114080, and George Washington University CDRF. applications,” Ultrasound Med. Biol. 26, 1177–1189.
Li, D. S., Kripfgans, O. D., Fabiilli, M. L., Brian Fowlkes, J., and Bull, J. L.
Aliabouzar, M., Kumar, K. N., and Sarkar, K. (2018). “Acoustic vaporiza- (2014). “Initial nucleation site formation due to acoustic droplet vapor-
tion threshold of lipid-coated perfluoropentane droplets,” J. Acoust. Soc. ization,” Appl. Phys. Lett. 104, 063703.
Am. 143, 2001–2012. Lo, A. H., Kripfgans, O. D., Carson, P. L., Rothman, E. D., and Fowlkes, J.
Apfel, R. E., and Holland, C. K. (1991). “Gauging the likelihood of cavita- B. (2007). “Acoustic droplet vaporization threshold: Effects of pulse dura-
tion from short-pulse, low-duty cycle diagnostic ultrasound,” Ultrasound tion and contrast agent,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control
Med. Biol. 17, 179–185. 54, 933–946.
Asami, R., and Kawabata, K. (2012). “Repeatable vaporization of optically Martin, A. L., Seo, M., Williams, R., Belayneh, G., Foster, F. S., and
vaporizable perfluorocarbon droplets for photoacoustic contrast enhanced Matsuura, N. (2012). “Intracellular growth of nanoscale perfluorocarbon
imaging,” in Proceedings of the 2012 IEEE International Ultrasonics droplets for enhanced ultrasound-induced phase-change conversion,”
Symposium (IUS), October 7–10, Dresden, Germany, pp. 1200–1203. Ultrasound Med. Biol. 38, 1799–1810.

J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al. 1115
Matsuura, N., Williams, R., Gorelikov, I., Chaudhuri, J., Rowlands, J., Reznik, N., Williams, R., and Burns, P. N. (2011). “Investigation of vapor-
Hynynen, K., Foster, S., Burns, P., and Resnik, N. (2009). “Nanoparticle- ized submicron perfluorocarbon droplets as an ultrasound contrast agent,”
loaded perfluorocarbon droplets for imaging and therapy,” in Proceedings Ultrasound Med. Biol. 37, 1271–1279.
of the 2009 IEEE International Ultrasonics Symposium (IUS), September Sarkar, K., Katiyar, A., and Jain, P. (2009). “Growth and dissolution of an
20–23, Roma, Italy, pp. 5–8. encapsulated contrast microbubble,” Ultrasound Med. Biol. 35,
Medwin, H., and Clay, C. S. (1997). Fundamentals of Acoustical 1385–1396.
Oceanography (Academic Press, New York). Sarkar, K., and Prosperetti, A. (1994). “Coherent and incoherent-scattering
Mercado, K. P., Radhakrishnan, K., Stewart, K., Snider, L., Ryan, D., and by oceanic bubbles,” J. Acoust. Soc. Am. 96, 332–341.
Haworth, K. J. (2016). “Size-isolation of ultrasound-mediated phase Sarkar, K., Shi, W. T., Chatterjee, D., and Forsberg, F. (2005).
change perfluorocarbon droplets using differential centrifugation,” “Characterization of ultrasound contrast microbubbles using in vitro
J. Acoust. Soc. Am. 139, EL142–EL148. experiments and viscous and viscoelastic interface models for
Miles, C. J., Doering, C. R., and Kripfgans, O. D. (2016). “Nucleation pressure encapsulation,” J. Acoust. Soc. Am. 118, 539–550.
threshold in acoustic droplet vaporization,” J. Appl. Phys. 120, 034903. Schad, K. C., and Hynynen, K. (2010). “In vitro characterization of perfluo-
Moncion, A., Arlotta, K. J., Kripfgans, O. D., Fowlkes, J. B., Carson, P. L., rocarbon droplets for focused ultrasound therapy,” Phys. Med. Biol. 55,
Putnam, A. J., Franceschi, R. T., and Fabiilli, M. L. (2016). “Design and 4933–4947.
characterization of fibrin-based acoustically responsive scaffolds for tissue Sheeran, P. S., Matsunaga, T. O., and Dayton, P. A. (2013). “Phase-transi-
engineering applications,” Ultrasound Med. Biol. 42, 257–271. tion thresholds and vaporization phenomena for ultrasound phase-change
Moncion, A., Lin, M., O’Neill, E. G., Franceschi, R. T., Kripfgans, O. D., nanoemulsions assessed via high-speed optical microscopy,” Phys. Med.
Putnam, A. J., and Fabiilli, M. L. (2017). “Controlled release of basic Biol. 58, 4513–4534.
fibroblast growth factor for angiogenesis using acoustically-responsive Sheeran, P. S., Matsunaga, T. O., and Dayton, P. A. (2014). “Phase change
scaffolds,” Biomaterials 140, 26–36. events of volatile liquid perfluorocarbon contrast agents produce unique
Olympus, N. (2006). “Ultrasonic transducers technical notes,” Technical acoustic signatures,” Phys. Med. Biol. 59, 379–401.
Brochure: Olympus NDT, Waltham, MA, 2.2. Sheeran, P. S., Wong, V. P., Luois, S., McFarland, R. J., Ross, W. D.,
Paul, S., Katiyar, A., Sarkar, K., Chatterjee, D., Shi, W. T., and Forsberg, F. Feingold, S., Matsunaga, T. O., and Dayton, P. A. (2011).
(2010). “Material characterization of the encapsulation of an ultrasound “Decafluorobutane as a phase-change contrast agent for low-energy extra-
contrast microbubble and its subharmonic response: Strain-softening inter- vascular ultrasonic imaging,” Ultrasound Med. Biol. 37, 1518–1530.
facial elasticity model,” J. Acoust. Soc. Am. 127, 3846–3857. Shpak, O., Verweij, M., Vos, H. J., de Jong, N., Lohse, D., and Versluis, M.
Paul, S., Nahire, R., Mallik, S., and Sarkar, K. (2014). “Encapsulated micro- (2014). “Acoustic droplet vaporization is initiated by superharmonic
bubbles and echogenic liposomes for contrast ultrasound imaging and tar- focusing,” Proc. Natl. Acad. Sci. 111, 1697–1702.
geted drug delivery,” Comput. Mech. 53, 413–435. Strohm, E. M., and Kolios, M. C. (2011). “Sound velocity and attenuation
Paul, S., Russakow, D., Rodgers, T., Sarkar, K., Cochran, M., and Wheatley, measurements of perfluorocarbon liquids using photoacoustic methods,”
M. A. (2013). “Determination of the interfacial rheological properties of a in Proceedings of the 2011 IEEE International Ultrasonics Symposium
Poly(DL-lactic acid)-encapsulated contrast agent using in vitro attenuation (IUS), October 18–21, Orlando, FL, pp. 2368–2371.
and scattering,” Ultrasound Med. Biol. 39, 1277–1291. Vlaisavljevich, E., Aydin, O., Yuksel Durmaz, Y., Lin, K. W., Fowlkes, B.,
Radhakrishnan, K., Bader, K. B., Haworth, K. J., Kopechek, J. A., ElSayed, M., and Xu, Z. (2015). “Effects of ultrasound frequency on
Raymond, J. L., Huang, S.-L., McPherson, D. D., and Holland, C. K. nanodroplet-mediated histotripsy,” Ultrasound Med. Biol. 41, 2135–2147.
(2013). “Relationship between cavitation and loss of echogenicity from Williams, R., Wright, C., Cherin, E., Reznik, N., Lee, M., Gorelikov, I.,
ultrasound contrast agents,” Phys. Med. Biol. 58, 6541–6563. Foster, F. S., Matsuura, N., and Burns, P. N. (2013). “Characterization of
Radhakrishnan, K., Holland, C. K., and Haworth, K. J. (2016). “Scavenging submicron phase-change perfluorocarbon droplets for extravascular ultra-
dissolved oxygen via acoustic droplet vaporization,” Ultrason. Sonochem. sound imaging of cancer,” Ultrasound Med. Biol. 39, 475–489.
31, 394–403. Zhang, P., Kopechek, J. A., and Porter, T. M. (2013). “The impact of vaporized
Rapoport, N. Y., Efros, A. L., Christensen, D. A., Kennedy, A. M., and nanoemulsions on ultrasound-mediated ablation,” J. Therap. Ultrasound 1, 2.
Nam, K.-H. (2009). “Microbubble generation in phase-shift nanoemul- Zhang, P., and Porter, T. (2010). “An in vitro study of a phase-shift nanoe-
sions used as anticancer drug carriers,” Bubble Sci. Eng. Technol. 1, mulsion: A potential nucleation agent for bubble-enhanced HIFU tumor
31–39. ablation,” Ultrasound Med. Biol. 36, 1856–1866.

1116 J. Acoust. Soc. Am. 145 (2), February 2019 Aliabouzar et al.

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