1. Introduction
Due to the nonlinear nature of biological media, the harmonic components of transmitted ultrasound are generated as the ultrasound propagates through the media [
1]. The energy of harmonics is nonlinearly proportional to transmitted ultrasound pressure; for example, the second harmonic energy is determined by the square of the energy in transmitted ultrasound [
2]. Therefore, the main lobe energy in the beam profile of transmitted ultrasound mainly contributes to the generation of the harmonics, thus leading to enhancing contrast resolution [
3,
4] because the levels of the side and grating lobes are considerably low in the harmonic beam profiles. Additionally, the main lobe width of a harmonic beam profile is narrower than that of a transmit beam profile and decreases as the harmonic order increases [
5]. Due to these properties, tissue harmonic imaging is capable of providing a higher spatial resolution than fundamental imaging, and the spatial resolution improves as the harmonic order increases. For these reasons, the second harmonic imaging of tissue is generally employed in modern ultrasound imaging systems. However, the superharmonic imaging is not yet a common imaging mode because a transducer with broad spectral bandwidth is not available for transmitting ultrasound and receiving its superharmonic components (i.e., higher than the second harmonic). Interleaved dual frequency array transducers can be the solution [
5,
6,
7], but the arrays suffer from a high level of grating lobes in fundamental imaging because the pitch of the elements generating fundamental ultrasound is doubled [
8]. Note that a transducer should provide fundamental images as well as harmonic images for accurate and efficient diagnosis.
For accurate diagnosis of atherosclerosis, it is also important that intravascular ultrasound (IVUS) images have high spatial and contrast resolutions. Active inflammation, severe stenosis, blood vessel remodeling, and a thin fibrous cap with a large lipid core are indispensable indications of vulnerable plaques [
9,
10], but current IVUS images do not have enough spatial and contrast resolutions to accurately detect the indicators. For example, the thickness of a thin fibrous cap (<65 μm) cannot be currently measured. In addition, lipid-rich plaque and blood vessel layers cannot be clearly visualized, which may cause the inaccurate of plaque composition assessment that is closely associated with plaque vulnerability [
10,
11]. This is so because conventional IVUS transducers have a flat (i.e., unfocused) aperture and center frequency in range of 20–40 MHz. To overcome the problem, focused IVUS transducers with a center frequency of 45–50 MHz have recently been developed [
12,
13], but the relatively high frequency may not be adequate to obtain the information about the overall morphological change of blood vessels due to atherosclerosis; vessel remodeling, lumen size, and wall morphology are closely related to the atheromatous plaque burden [
10]. Note that ultrasound imaging depth decreases as operating frequency increases. For this end, dual-frequency IVUS transducers are the best choice to achieve both high spatial resolution and deep imaging depth: low frequency element for general IVUS imaging and high frequency element for superficial microstructure imaging [
14,
15,
16]. Additionally, those IVUS transducers facilitate the superharmonic imaging of tissue, thus further improving spatial and contrast resolutions. However, the structure in which two elements are attached back and forth [
14,
15] cannot be used for tissue harmonic imaging because the harmonics generated in tissue cannot be received. Although the arrangement of two elements side by side in the elevation direction is suitable for tissue harmonic imaging, the simple arrangement of two pre-focused elements is not the case [
16]. In this case, harmonic signals cannot be received effectively because the focal positions of the two elements are different. For tissue harmonic imaging, therefore, dual-frequency elements arranged side by side in the elevation direction should have a spherical shape so that their focal positions can be the same; recently, we have reported a dual-frequency oblong-shaped-focused IVUS transducer for the second harmonic imaging of tissue [
17], in which the two outer 35-MHz elements play a role of transmitters, and the center 70-MHz element serves as the receiver of the second harmonic signals.
For superharmonic imaging in conjunction with ultrasound contrast agents, dual-frequency IVUS transducers in which two elements were stacked vertically were developed [
18,
19,
20]. The lower layer element is responsible for resonating injected microbubbles that generate superharmonics. For this, the center frequency of the lower layer is in the range of 2–10 MHz. Since the upper layer element receives the superharmonics, the center frequency is 30 MHz. These transducers may be suitable for obtaining information about the proliferation of the adventitial vasa vasorum that is closely related to early atherosclerotic plaque [
21], but not for the superharmonic imaging of tissue. The lower layer has a relatively large flat aperture such as 0.6 × 3 mm
2, so that the natural focal depth is far beyond the imaging region of interest. This means that the size and shape of the element are inappropriate for tissue imaging. Additionally, the operating frequencies are too low to acquire high quality images. If the center frequencies are increased for enhancing spatial resolution, the upper layer element causes severe distortion and attenuation of the ultrasound generated by the lower layer element. This hampers the transmission of ultrasound energy high enough to produce superharmonics within tissue.
In this paper, we report a recently developed dual-element IVUS transducer for the third harmonic imaging of tissue. The two elements are arranged side by side in the elevation direction and spherically shaped with a radius of 2.5 mm. This configuration was adopted for optimal generation and reception of harmonics as well as avoiding the misalignment of the beam profiles of the two elements in the imaging region of interest. The two elements were designed to have center frequencies of 35 MHz for ultrasound transmission and 105 MHz for the reception of the third harmonic, respectively. High efficient transmission and reception of ultrasound require electrical impedance matching between a transducer and an imaging system (or signal wires) [
22]. At a given element size, the 105-MHz element inevitably has a much lower electrical impedance than the 35-MHz element if the same piezoelectric material is used [
8]. To avoid this, lithium niobate (LiNbO
3) was used for the 105-MHz element, whereas PZT-5H served as the active material of the 35-MHz element. Note that the clamped dielectric constant of LiNbO
3 is lower than that of PZT-5H. The dual-element transducer was designed using a KLM model based simulator, and its characteristics and imaging performance were evaluated.
3. Performance Evaluation and Discussion
The electrical impedance of the fabricated dual-element focused transducer was measured using an impedance analyzer (HP4294A, Agilent Corp., Santa Clara, CA, USA) with z-probe attachment as shown in
Figure 7. In the case of the 35-MHz element, the electrical impedance was measured at a magnitude of 46.7 Ω and a phase angle of −72.12° at 35 MHz. The 105-MHz element had the electrical impedance with a magnitude of 34.9 Ω and a phase angle of −79.85° at 105 MHz. Since a pulser/receiver system (UT340, UTEX Scientific Instruments Inc., Mississauga, ON, Canada) was used for pulse-echo tests and imaging performance evaluation, the electrical impedance of the 35-MHz element was considered to be well matched to the output impedance of the system, i.e., 50 Ω. In contrast, the 105-MHz element had the electrical impedance lower than the system’s output impedance, but this impedance mismatching was not a serious problem for operating the element. The problem was the phase distortion of the ultrasonic wave generated by the element, which led to elongating pulse length and thus degrading axial resolution. Therefore, we conducted electrical impedance matching using a series capacitor of 24 pF, a shunt capacitor of 13 pF, and a shunt inductor of 33 nH for minimizing the phase distortion. After the impedance matching, the electrical impedance magnitude and phase angle were changed to 34.9 Ω and −16.99°. For pulse-echo tests, the fabricated transducer was immersed into a container filled with a deionized water. A polished steel target playing a role of a reflector was placed in the container. The position of the transducer was adjusted using a motorized stage (SGSP26-100, SIGMAKOKI Co. Ltd., Tokyo, Japan) so that the focal point is located on the reflector. The pulser/receiver system was used to excite the elements and to receive the echoes from the reflector. The received echoes were recorded using an oscilloscope (DPO7054, Tektronics Inc., Beaverton, OR, USA) and used to calculate the center frequency and −6 dB fractional bandwidth of each element in MATLAB (Mathworks Inc., Natick, MA, USA). From the pulse-echo tests, it was found that the 35-MHz element had a center frequency of 36 MHz and a −6 dB fractional bandwidth of 56% (i.e., 26.1 to 46.6 MHz) as shown in
Figure 8, which is in good agreement with the simulation results. However, the center frequency of the 105-MHz element was 112 MHz, which is higher than a prediction value of 107 MHz. In addition, the −6 dB fractional bandwidth was narrowed to 29% (i.e., 95.4 to 128 MHz). At the 105-MHz element, this discrepancy between the simulation results and the measurements may stem from the thickness of LiNbO
3 being thinner than the target thickness (i.e., 28 μm). This is so because the lapping error leads to a considerable change in the center frequency of the 105-MHz element due to the very thin thickness of LiNbO
3.
The crosstalk between the two elements was measured. For this, the IVUS transducer was immersed into the water container, and the reflector was replaced by an acoustic absorber. A five-cycle sinusoidal burst signal with 5 V
p-p was generated by a function generator (AFG3102, Tektronix Inc., Beaverton, OR, USA) and was sent to both 35-MHz element and oscilloscope with an input impedance set to 1 MΩ. At the same time, the voltage induced by the sinusoidal signal at the 105-MHz element was measured using the same oscilloscope. The frequency of the sinusoidal signal was changed from 10 to 120 MHz at 5 MHz intervals and the ratio of the output voltage of the 105-MHz element to the input voltage of the 35-MHz element was computed (see the solid line in
Figure 9); the maximum crosstalk was −29.9 dB at 110 MHz. In addition, the crosstalk was measured from the 35-MHz element while exciting the 105-MHz element (see the dashed line) and the maximum crosstalk in this case was −29.4 dB at 115 MHz. Note that the crosstalk measured in this study is a combination of acoustic and electrical crosstalk. At the design stage, acoustic crosstalk was expected to be ignored because the 5 min epoxy filled kerf for acoustically and electrically isolating the two elements had a relatively large value of 0.1 mm, compared to those of conventional arrays. However, it was anticipated that electrical crosstalk would dominate overall crosstalk because there were two parallel wires inside the brass housing with a diameter of 1 mm. This can be confirmed by the measured crosstalk that increases with the frequency used for the measurement as shown in
Figure 9. This is so because a pair of coupled lines generate electrical crosstalk with each other, and the crosstalk is linearly proportional to frequencies below a few hundred MHz [
26]. Nevertheless, the crosstalk of the fabricated dual-element IVUS transducer is acceptable for imaging because the crosstalk of conventional ultrasound arrays typically has a value between −28 and −32 dB [
27].
The imaging performance of the developed transducer was also evaluated through tissue-mimicking phantom tests. For this, the tissue-mimicking phantom was constructed [
28]; a deionized water of 250 mL in a beaker was heated to 60 °C on a hot plate. The mixture of an agar powder (A9799, Sigma-Aldrich Co. Ltd., St. Louis, MO, USA) of 8.0 g and a silicon dioxide powder (S5631, Sigma-Aldrich Co. Ltd., St. Louis, MO, USA) of 5.0 g were prepared and poured into the beaker while stirring the water with a rod. The mixture was cooled down at room temperature and cured in a refrigerator for 12 h. For insertion of the transducer, a hole was created in the center of the phantom. In addition, two 26 G 1/2 needles were used to create a lesion-mimicking area at the surface of the phantom; the diameter of each hole was 0.45 mm. To acquire image data, the developed transducer was inserted into the hole of the fabricated phantom in the deionized-water-filled container on the rotary stage (SGSP 160-YAW, SIGMAKOKI Co. Ltd., Tokyo, Japan) as shown in
Figure 10. For the third harmonic imaging, the pulser/receiver system was used to excite the 35-MHz element and receive ultrasound by using the 105-MHz element. Note that 35- and 105-MHz fundamental images were also acquired using the pulser/receiver system. The received ultrasound was digitized and recorded using a Gage card (CS12502, Gage Applied Technologies Inc., Montreal, QC, Canada) in a personal computer. For one image, 1000 scanlines were acquired by rotating the stage in 0.36° increments. The scanning process was controlled by a program written in LabView (National Instrument, Austin, TX, USA). All signal processing for image construction with the recorded image data was conducted in MATLAB. For DC cancelling and noise reduction [
29], a 129-tap band pass filter was used: cutoff frequencies of 21 and 52 MHz for the 35-MHz fundamental images and 90 and 133 MHz for both 105-MHz fundamental and third harmonic images. The images were logarithmically compressed with a dynamic range of 30 dB. Additionally, the spatial resolution of the fabricated transducer was measured using a 25-μm wire under the experimental setup. For this, the wire was placed at the focal point of the transducer, i.e., 2.5 mm before acquiring the 35-MHz fundamental, 105-MHz fundamental, and third harmonic scanlines. From the scanlines, the axial and lateral beam profiles were obtained after envelope detection [
30], and their −6 dB beam widths were measured.
As shown in
Figure 11, the −6 dB axial and lateral beam widths of the 105-MHz fundamental and the third harmonic beam profiles were equal: an axial beam width of 25 μm and a lateral beam width of 46 μm. These were superior to the −6 dB axial and lateral beam widths of the 35-MHz fundamental beam profile, i.e., 40 and 153 μm, respectively. This is an obvious result because ultrasound frequency determines the widths of axial and lateral beam profiles directly related to the spatial resolution of ultrasound images. However, the level of side lobes associated with contrast resolution as well as spatial resolution was the lowest in the beam profile of the third harmonic, even though the beam profile was generated by the beam profile of the 35-MHz element that had the highest level of side lobes. This result demonstrates the superiority of the harmonic imaging in contrast resolution, which is possible because the energy of harmonics is nonlinearly proportional to transmitted ultrasound pressure and thus the main lobe energy in the beam profile of transmitted ultrasound mainly contributes to the generation of the harmonics.
Figure 12 shows the 35-MHz and 105-MHz fundamental images and the third harmonic image of the tissue-mimicking phantom. It was observed that the 35-MHz fundamental image had a deeper imaging depth than other images, but the speckle pattern appeared to spread along the arc as the imaging depth increased and the lesion-mimicking area in the phantom were not clearly seen due to low spatial resolution, compared to the others. In contrast, the 105-MHz fundamental image showed the conspicuous speckle pattern and clear lesion-mimicking area indicated by white arrows in
Figure 12 due to its high spatial resolution. However, the frequency-dependent attenuation of ultrasound limited the imaging depth to less than 1.0 mm from the surface. The third harmonic image had a spatial resolution similar to the 105-MHz fundamental image, whereas its imaging depth was deeper than that of 105-MHz fundamental image, i.e., up to 1.4 mm. This result is reasonable because traveling distance of the fundamental ultrasound is twice the traveling distance of the third harmonic, so that the third harmonic experiences less attenuation.
For tissue harmonic imaging, the spectrum of a transducer is typically divided into two sub-bands: one for transmission and another for harmonic reception. This causes lowering the axial resolution of both fundamental and harmonic images. Additionally, the complete separation of fundamental and harmonic signals is very challenging because their spectra are overlapped although pulse inversion and nonlinear chirp excitation can be used to overcome the problem partially [
31,
32,
33]. Given these facts, the wide spectral bandwidth of a transducer is essential for high quality tissue harmonic imaging. However, it is practically impossible to construct a transducer with a broad bandwidth enough to carry out the third harmonic imaging of tissue. This is especially true for IVUS transducers because of their high operating frequency and limited selection of backing and acoustic matching materials due to the structural restriction. Even if possible, a severe degradation of spatial resolution is inevitable because the bandwidth should be divided into three sub-bands. As shown in
Figure 13, the developed dual-element focused transducer can solve those problems. For ultrasound transmission, the full bandwidth of the 35-MHz element can be exploited, so that no degradation of spatial resolution occurs in fundamental images. This full-bandwidth signal contributes to the generation of the third harmonic, which is received by the 105-MHz element without (or at a minimum) loss of the harmonic information. Note that the frequency spectral characteristics of the 105-MHz element is similar to a band-pass filter, so that the signal separation is not necessary. Another advantage of the developed transducer is the simultaneous acquisition of co-registered 35- and 105-MHz fundamental images, which enables us to examine the overall morphological change of blood vessels and the near vessel layers such as fibrous cap with high spatial resolution at the same time if necessary.