1648 Vol. 49, No.

7 / 1 April 2024 / Optics Letters Letter

Sub-Nyquist sampling-based high-frequency

photoacoustic computed tomography
Songde Liu,1,2,3 Chenxi Zhang,2 Junyi Zhang,4 Xingyang Liu,2 Benpeng Zhu,5,6 AND
Chao Tian1,2,3,∗
1
Department of Anesthesia, the First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology
of China, Hefei, Anhui 230026, China
2
School of Engineering Science, University of Science and Technology of China, Hefei, Anhui 230026, China
3
Institute of Artificial Intelligence, Hefei Comprehensive National Science Center, Hefei, Anhui 230088, China
4
Institute of Advanced Technology, University of Science and Technology of China, Hefei, Anhui 230088, China
5
School of Integrated Circuits, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei
430074, China
6
benpengzhu@hust.edu.cn
*ctian@ustc.edu.cn

Received 11 December 2023; revised 14 February 2024; accepted 3 March 2024; posted 5 March 2024; published 18 March 2024

High-frequency (greater than 30 MHz) photoacoustic com-
puted tomography (PACT) provides the opportunity to
reveal finer details of biological tissues with high spatial
resolution. To record photoacoustic signals above 30 MHz,
sampling rates higher than 60 MHz are required according
to the Nyquist sampling criterion. However, the highest sam-
pling rates supported by existing PACT systems are typically
within the range of 40–60 MHz. Herein, we propose a novel
PACT imaging method based on sub-Nyquist sampling. The
results of numerical simulation, phantom experiment, and
in vivo experiment demonstrate that the proposed imaging
method can achieve high-frequency PACT imaging with a
relatively low sampling rate. An axial resolution of 22 µm is
achieved with a 30-MHz transducer and a 41.67-MHz sam-
pling rate. To the best of our knowledge, this is the highest
axial resolution ever achieved in PACT based on a sampling
rate of not greater than 60 MHz. This work is expected to
provide a practical way for high-frequency PACT imaging
with limited sampling rates.
© 2024 Optica Publishing Group under the terms of the Optica Open
Access Publishing Agreement
https://doi.org/10.1364/OL.515650
Photoacoustic (PA) computed tomography (PACT) is a fast-
evolving biomedical imaging modality in the last two decades
[1]. Based on the PA effect, PACT can achieve high spa-
tiotemporal resolution at large imaging depths and thus has
vast applications in a range of biomedical fields [2], such
as neurology [3], oncology [4], rheumatology [5], surgical
navigation [6], and other areas of medical imaging prob-
lems. High-resolution PACT provides the opportunity to reveal
finer details of biological tissues [7–9]. The spatial resolution
of a PACT system can be affected by multiple system fac-
tors and is closely related to the characteristics of ultrasonic
detectors, including the center frequency, bandwidth, aperture
size, and view angle [10]. To enhance the spatial resolution
and thus improve the image quality of PACT, one can sim-
ply increase the center frequency of ultrasonic detectors [9].
According to the Nyquist sampling criterion, high-frequency
PACT imaging demands data acquisition (DAQ) systems with
analog-to-digital converters (ADC) of high sampling rates.
In practice, the highest sampling rates supported by existing
PACT systems are typically within the range of 40–60 MHz
[11], which leads to a Nyquist frequency of not greater than
30 MHz.
To achieve high-frequency PACT imaging with band-
widths covering frequencies above 30 MHz, researchers usually
employ DAQ systems with sampling rates higher than 60 MHz
[8,12–14]. However, such DAQ systems are usually not cost-
effective. Inspired by a delayed-excitation DAQ method previ-
ously reported for high-frequency ultrasound imaging [15], Fu
and Jokerst proposed a new method called interleave-sampled
PA imaging that enables high-frequency PACT imaging with a
relatively low sampling rate, e.g., a 41.67-MHz sampling rate
with a 30-MHz transducer [16]. The proposed method requires
two acquisitions that are precisely shifted with each other by half
of the sampling period and are sampled at the same sampling rate
(e.g., 41.67 MHz). Interleaving these two acquisitions forms a
virtual acquisition that is equivalent to the acquisition sampled at
a doubled sampling rate (e.g., 83.33 MHz). The authors achieved
a 63-µm axial resolution and a 91-µm lateral resolution via the
proposed imaging method.
In this Letter, we propose another novel imaging method based
on sub-Nyquist sampling that can also achieve high-frequency
PACT imaging with a relatively low sampling rate. Sub-Nyquist
sampling, also known as bandpass sampling, allows for sampling
bandpass signals with a sampling rate less than twice the highest
frequency content [17] and has been widely used in ultrasound
imaging for reducing computational complexity [18]. The idea
of the proposed method is based on the fact that the frequency
response of high-frequency ultrasonic detectors is usually band-
limited.

0146-9592/24/071648-04 Journal © 2024 Optica Publishing Group

 Letter
Fig. 1. Schematic illustrating the basic principle of the pro-
posed sub-Nyquist sampling-based high-frequency PACT imaging
method. (a) Flow chart of the proposed method. (b) Original analog
signal. (c) Filtered analog signal. (d) Replicated digital signal with
spectral inversion. (e) Recovered digital signal.
Suppose we have a bandpass signal with a center frequency
of f c and a bandwidth of B. According to the theory of sub-
Nyquist sampling [17], we can sample the bandpass signal
without aliasing at a rate of f s that is constrained to
2fc − B 2fc + B
≥ fs ≥ , (1)
m m+1
where m is an arbitrary, positive integer ensuring that f s ≥ 2B.
We can rearrange Eq. (1) as
2fc 2fc
B≤ ≤ . (2)
2m + 1 3
Smaller bandwidth brings larger m and thus smaller f s . Since
bandwidth is essential for high-quality PACT imaging [9], we
should choose smaller m, e.g., m = 1 in this Letter. In this case,
the highest bandwidth supported by sub-Nyquist sampling is
2f c /3, and the corresponding f s is 4f c /3. Figure 1(a) illustrates
the basic principle of the proposed sub-Nyquist sampling-
based high-frequency PACT imaging method (m = 1, B = 2f c /3,
f s = 4f c /3). Firstly, we employ a bandpass filter (passband: [2/3f c ,
4/3f c ]) to eliminate the noise and undesired spectra outside the
band of interest [Figs. 1(b) and 1(c)]. Secondly, we sample the fil-
tered signal at 4f c /3. The desired spectrum in the high-frequency
band ([2f c /3, 4f c /3]) is replicated in the low-frequency band ([0,
2f c /3]) with spectral inversion after ADC digitization [Fig. 1(d)].
Thirdly, we fold the replicated spectrum back into the original
high-frequency band ([2f c /3, 4f c /3]) along 2f c /3 [Fig. 1(e)]. The
sampling rate is doubled in this step. The recovered spectrum
shown in Fig. 1(e) is identical to the original spectrum of inter-
est shown in Fig. 1(c). Finally, the recovered signal is used for
PACT image reconstruction. Specifically, the third step (recov-
ery of aliased signal) can be implemented as follows. Suppose
F(ω) is the Fourier spectrum of the aliased digital signal, f (t).
To fold F(ω) back into the original high-frequency band, we
Vol. 49, No. 7 / 1 April 2024 / Optics Letters 1649
Fig. 2. Numerical simulation confirming the feasibility of the
proposed imaging method. (a) Schematic of the simulation setup
showing a 20-µm microsphere placed 8 mm from a linear transducer
array. (b) Ground truth for the simulation obtained with Config. 6.
(c)–(g) Imaging results obtained with Configs. 1 to 5. (h) Summation
of (c) and (d). (i) and (j) Axial and lateral resolution comparisons.
construct another spectrum, G(ω), as
{︃
F(4fc /3 − |ω|), if 2fc /3<|ω|<4fc /3
G(ω) = (3)
0, otherwise.
Taking the inverse Fourier transform of G(ω) gives the recovered
digital signal g(t), the sampling rate of which is twice that of f (t).
It is critical to ensure that the original analog signals are
filtered with an analog bandpass filter before being sampled
by the ADC to eliminate the noise and undesired spectra out-
side the band of interest. Otherwise, these noise and undesired
spectra would be added to the replicated spectrum after digitiza-
tion. Under such circumstances, the original spectrum of interest
would be contaminated and could not be recovered accurately.
To demonstrate the performance of the proposed imaging
method, numerical simulation, phantom experiment, and in vivo
experiment were conducted. The numerical simulation was per-
formed by imaging a microsphere. Considering that smaller
microspheres yield higher frequency content, the diameter of
the microsphere was set to 20 µm to represent a point target
while matching the detection bandwidth of the transducer to
maximize detection sensitivity as much as possible. An ana-
lytical model [19] was employed for generating PA signals of
the microsphere, which were then received by a 30-MHz linear
transducer array with a bandwidth of 20 MHz. The microsphere
was placed 8 mm from the transducer array, as shown in Fig. 2(a).
The received PA signals were first digitized at 1 GHz to serve
as the original analog signals. To compare the performance of
the proposed method with other methods, the digitized signals
were separately filtered with multiple filtering configurations
and down-sampled to different sampling rates (Table 1). Herein,
we suppose the highest sampling rate supported by the DAQ is
62.5 MHz. Sampling rates of 83.33 and 125 MHz were used for
simulating interleaved sampling based on 41.67 and 62.5 MHz,
 1650 Vol. 49, No. 7 / 1 April 2024 / Optics Letters
Table 1. Sampling Configurations for Performance
Comparison
Configuration Sampling Highpass Filter Lowpass Filter
Rate Cutoff Freq. Cutoff Freq.
(MHz) (MHz) (MHz)
1 41.67 20 35
2 41.67 - 20
3 62.5 - 30
4 83.33 - 35
5 125 - 50
6 250 - -
respectively. A highpass filter and a lowpass filter were com-
bined to act as a bandpass filter for Config. 1. For Configs. 2 to
5, a lowpass filter configured with different cutoff frequencies
was employed to avoid aliasing resulting from frequency con-
tent above half of the sampling rates. Varying sampling rates for
Configs. 2 to 5 leads to varying cutoff frequencies of the low-
pass filter. The cutoff frequencies and spectral responses of the
above filters can be found in Table 1 and Fig. S1 (Supplement 1).
PACT image reconstruction was performed based on the filtered
back-projection algorithm [19].
Figure 2(b) shows the imaging result obtained based on Con-
fig. 6. Due to the limited detector bandwidth [see Fig. S2(f),
Supplement 1] and view angle [9], the reconstructed micro-
sphere shown in Fig. 2(b) is not round-shaped. Nevertheless,
Fig. 2(b) is regarded as the ground truth in this simulation. Fig-
ures 2(c)–2(g) are the imaging results obtained with Configs. 1
to 5. Negativity artifacts shown in Figs. 2(b)–2(g) result from the
limited detector bandwidth and view angle [20]. Intensity pro-
files along the red dashed lines shown in Fig. 2(b) are plotted in
Figs. 2(i) and 2(j) with the FWHM (full width at half maximum)
quantified as axial (z axis) and lateral (x axis) resolution. Due
to spectral aliasing and the loss of low-frequency content [see
Fig. S2(a), Supplement 1] [9], Fig. 2(c) shows clearer ringing
artifacts (white arrow) when compared with the ground truth.
The hollow structure of the microsphere shown in Fig. 2(d), the
moderately broadened microsphere shown in Fig. 2(e), and the
mildly broadened microsphere shown in Fig. 2(f) both result
from spectral aliasing and the loss of high-frequency content
[see Figs. S2(b)–S2(d), Supplement 1]. Considering that Con-
figs. 5 and 6 have similar patterns of frequency content [see
Figs. S2(e) and S2(f), Supplement 1], Fig. 2(g) shows similar
imaging result to the ground truth. Figure 2(c) is sharper than
Fig. 2(e) owing to the sampling of a higher frequency con-
tent. The result obtained based on the proposed imaging method
[Fig. 2(c)] is as sharp as those obtained based on the interleaved
imaging method [Figs. 2(f) and 2(g)]. However, Fig. 2(c) suf-
fers from severe ringing artifacts that can be well suppressed by
adding Fig. 2(d) to Fig. 2(c), which yields Fig. 2(h). The above
findings are well supported by the axial and lateral resolution
comparisons shown in Figs. 2(i) and 2(j).
A fine hair (diameter: 10–20 µm) was embedded in an agar
phantom and imaged to evaluate the performance of the pro-
posed method in practice. The specifications of the imaging
system can be found in [6] and will not be detailed here.
The highest sampling rate supported by the imaging system
is 62.5 MHz. A commercial 256-channel linear transducer array
(MS400; Visualsonics, Inc., Canada) with a center frequency of
30 MHz and a bandwidth of 20 MHz was used for signal receiv-
ing. The imaging settings are the same as those described in the
Letter
Fig. 3. Phantom experiment of a fine hair evaluating the perfor-
mance of the proposed method in practice. (a)–(e) Imaging results
obtained with Configs. 1 to 5. (f) Summation of (a) and (b). (g)
Axial intensity profiles along the red dashed line shown in (e).
numerical simulation. Figures 3(a)–3(e) are the imaging results
obtained with Configs. 1 to 5. As expected, the proposed imag-
ing method [Fig. 3(a)] provides finer structure but worse ringing
artifacts (white arrows) compared with the conventional imag-
ing method [Fig. 3(c)]. The ringing artifacts shown in Fig. 3(a)
are well suppressed by adding Fig. 3(b) to Fig. 3(a), as shown in
Fig. 3(f). The interleaved imaging method also provides shaper
results [Figs. 3(d) and 3(e)] compared with the conventional
imaging method [Fig. 3(c)]. Figure 3(a) shows a smaller FWHM
[Fig. 3(g)] and severe ringing artifacts, both resulting from spec-
tral aliasing and the loss of low-frequency content, compared
with Fig. 3(d). On the contrary, Fig. 3(b) shows a larger FWHM
[Fig. 3(g)], resulting from spectral aliasing and the loss of high-
frequency content, compared with Fig. 3(d). Theoretically, the
combination of Configs. 1 and 2 should cancel each other out
and produce a similar FWHM [Fig. 3(f)] as that obtained with
Config. 4 [Fig. 3(d)]. However, due to spectral aliasing, Fig. 3(f)
shows a slightly smaller FWHM. The above findings are well
supported by the axial intensity profiles [Fig. 3(g)] and the digital
spectra (see Fig. S3, Supplement 1).
Finally, we conducted an in vivo experiment on a mouse ear to
explore the potential of the proposed method in imaging complex
biological tissues. Approval of all ethical and experimental pro-
cedures and protocols was granted by the Institutional Animal
Care and Use Committee (IACUC) of the University of Sci-
ence and Technology of China under No. USTCACUC1803065.
Blood vessels of the mouse ear shown in Fig. 4(a) were imaged
based on Configs. 1 to 5, with results presented in Figs. 4(b)–4(f).
As expected, the proposed imaging method [Fig. 4(b)] provides
higher spatial resolution but suffers from severe ringing arti-
facts when compared with the conventional imaging method
[Fig. 4(d)]. The ringing artifacts shown in Fig. 4(b) are well sup-
pressed by adding Fig. 4(c) to Fig. 4(b), as shown in Fig. 4(g).
The performance of the combined imaging method [Fig. 4(g)]is
comparable to that of the interleaved imaging method [Figs. 4(e)
and 4(f)]. The findings are well supported by the characterized
FWHM values [Figs. 4(h) and 4(i)] of the blood vessel shown
in insets and the digital spectra (see Fig. S4, Supplement 1).
Due to hardware limitation, the cutoff frequency of the
highpass filter in Config. 1 was configured as 20 MHz, not
20.83 MHz. For consistency, the 20-MHz cutoff frequency
was used in both the simulation and experiments. A specially
designed numerical simulation shows that the impact of this
tiny deviation (20.83 MHz versus 20 MHz) on imaging results
 Letter
Fig. 4. In vivo experiment of a mouse ear exploring the potential
of the proposed method in imaging complex biological tissues.
(a) Photograph of the mouse ear showing lots of microvessels.
(b)–(f) Imaging results of blood vessels (white arrows) obtained
with Configs. 1 to 5. (g) Summation of (b) and (c). Insets are the
zoomed-in images of the vessel in the yellow dashed boxes. (h) and
(i) Intensity profiles of the blood vessel shown in insets along z axis
(h) and x axis (i) with FWHM values characterized.
is insignificant (see Fig. S5, Supplement 1). In practice, the ana-
log filters are usually nonideal, having limited suppression ratios
beyond the band of interest, which will lead to spectral aliasing
and affect the imaging quality to some extent. Another specially
designed numerical simulation indicates that high-performance
analog filters are expected to reduce imaging artifacts resulting
from the limited suppression ratios of the nonideal filters (see
Fig. S6, Supplement 1).
The proposed method has the same level of complexity as the
interleaved sampling method when two acquisitions are neces-
sary (Configs. 1 and 2). However, under some circumstances, the
proposed method may be superior to the interleaved sampling
approach. Specifically, the former only needs one acquisition
(Config. 1) to capture a high-frequency content, while the latter
principally needs two. Although the results shown here suggest
that two acquisitions are required to suppress ringing artifacts,
we believe there may be scenarios (e.g., machine learning is used
to suppress the ringing artifacts resulting from the loss of low-
frequency components) where only one acquisition is sufficient
in the future.
Herein, we demonstrate the concept based on a 30-MHz trans-
ducer. But actually, the proposed method can be used for signals
with cutoff frequencies beyond 40 MHz when appropriate band-
pass filters are available. High-frequency PACT primarily targets
small structures (e.g., microvessels) whose frequency content
matches the detection bandwidth of the transducer. Most low-
frequency content produced by large structures will be lost due
to the limited detection bandwidth of the transducer, but not the
inappropriate setting of the sampling configuration. The loss
of low-frequency content is also inevitable for the interleaved
Vol. 49, No. 7 / 1 April 2024 / Optics Letters 1651
sampling method and only impacts the image quality of large
structures, not small structures.
In summary, we proposed a novel imaging method based on
sub-Nyquist sampling to achieve high-frequency PACT imag-
ing with a relatively low sampling rate. The results show that
the proposed imaging method provides better spatial resolution
when compared to the conventional imaging method. It is nec-
essary to add another acquisition that samples a low-frequency
content to the proposed acquisition to suppress ringing artifacts
induced by the proposed imaging method. The performance of
the combined imaging method is comparable to that of the inter-
leaved imaging method. We achieve an axial resolution of 22 µm
based on a relative low sampling rate (i.e., 41.67 MHz) with the
proposed imaging method. To the best of our knowledge, this is
the highest axial resolution ever achieved in PACT based on a
sampling rate of not greater than 60 MHz. This work is expected
to provide a practical way for high-frequency PACT imaging
with limited sampling rates.
Funding. National Key Research and Development Program of China
(2022YFA1404400); National Natural Science Foundation of China
(12174368, 61705216, 62122072); Anhui Provincial Department of Science
and Technology (18030801138, 202203a07020020); University of Science
and Technology of China (YD2090002015); Institute of Artificial Intelli-
gence at Hefei Comprehensive National Science Center (23YGXT005).
Disclosures. The authors declare no conflicts of interest.
Data availability. Data underlying the results presented in this paper are
not publicly available at this time but may be obtained from the authors upon
reasonable request.
Supplemental document. See Supplement 1 for supporting content.
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