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This protocol shows how to apply ultrafast ultrasound Doppler imaging to quantify blood flows. After a 1 s long acquisition, the experimenter has access to a movie of the full field of view with axial velocity values for each pixel every ≈0.3 ms (depending on the ultrasound time of flight).
The pulsed-Doppler effect is the main technique used in clinical echography to assess blood flow. Applied with conventional focused ultrasound Doppler modes, it has several limits. Firstly, a finely tuned signal filtering operation is needed to distinguish blood flows from surrounding moving tissues. Secondly, the operator must choose between localizing the blood flows or quantifying them. In the last two decades, ultrasound imaging has undergone a paradigm shift with the emergence of ultrafast ultrasound using unfocused waves. In addition to a hundredfold increase in framerate (up to 10000 Hz), this new technique also breaks the conventional quantification/localization trade-off, offering a complete blood flow mapping of the field of view and a simultaneous access to fine velocities measurements at the single-pixel level (down to 50 µm). This data continuity in both spatial and temporal dimensions strongly improves the tissue/blood filtering process, which results in an increase sensitivity to small blood flow velocities (down to 1 mm/s). In this method paper, we aim to introduce the concept of ultrafast Doppler as well as its main parameters. Firstly, we summarize the physical principles of unfocused wave imaging. Then, we present the Doppler signal processing main steps. Particularly, we explain the practical implementation of the critical tissue/blood flow separation algorithms and on the extraction of velocities from these filtered data. This theoretical description is supplemented by in vitro experiences. A tissue phantom embedding a canal with flowing blood-mimicking fluid is imaged with a research programmable ultrasound system. A blood flow image is obtained and the flow characteristics are displayed for several pixels in the canal. Finally, a review of in vivo applications is proposed, showing examples in several organs such as carotids, kidney, thyroid, brain and heart.
Ultrasound imaging is one of the most commonly used imaging techniques in clinical practice and research activities. The combination of ultrasound wave emission in the biological tissues followed by the recording of the backscattered echoes allows the reconstruction of anatomical images, the so-called “B-Mode”. This method is perfectly adapted for soft tissue imaging, such as biological tissues, which typically permit the penetration of ultrasound over several centimeters, with a propagation speed of ≈1540 m/s. Depending on the center frequency of the ultrasound probe, images with a resolution from 30 µm to 1 mm are obtained. Furthermore, it is well known that the motion of an acoustic source, affects the physical characteristics of the associated waves. Particularly, the link between the frequency shifts of a wave relative to the speed of its source is described as the Doppler effect1, whose simplest manifestation is the changing siren’s pitch of a moving ambulance. Ultrasound imaging has long used this physical effect to observe the moving red blood cells2, and it proposes a variety of imaging modes commonly labelled “Doppler imaging”. These modes enable the assessment of blood flows in very different applications and organs, such as brain, heart, kidney or peripheral arteries.
Remarkably, most of the currently available ultrasound systems rely on the same technology, referred to as conventional ultrasound. The underlying principles are the following: an acoustic beam insonifies the field of view and is swept along the ultrasound transducer aperture. For each position of the beam, the echoes are recorded and converted into a line of the final image. By progressively moving the beam along the transducer, the whole field of view can be imaged line-per-line (Figure 1, left panel). This strategy was well adapted to the electrical constraints and computing power prevailing until the beginning of the 21st century. Nonetheless, it has several drawbacks. Among these, the final framerate is limited to a few hundred images per second by the beam scanning process. In terms of blood flow, this relatively low framerate affects the maximum flow velocities that can be detected, which is dictated by the sampling criteria of Shannon-Nyquist3. Moreover, conventional Doppler must deal with a complex tradeoff. In order to assess the blood flow velocity in a particular region of interest (ROI), several echoes coming from that ROI have to be successively recorded. This implies that the ultrasound beam is temporarily maintained in a fixed position. The longer the echo ensemble, the better the velocity estimation will be for that ROI. However, to produce a complete image of the field of view, the beam must scan the medium. Therefore, one can sense the conflict between these two constraints: holding the beam to precisely assess the velocity along one line, or moving the beam to produce an image. The different conventional Doppler modes (i.e., Color Doppler or Pulse Wave Doppler) directly reflect this tradeoff. Typically, the Color Doppler produces a low-fidelity flow map used for localizing the vessels4, and the Pulse Wave Doppler is then used to accurately quantify the flow in a previously identified vessel5.
These two limitations (low framerate and localization/quantification tradeoff) are overcome with very high-framerate emerging techniques. Among these, the synthetic aperture approach6 or the multiline transmit technique can be cited7. In this study, we focus on the so-called Ultrafast ultrasound method. Introduced two decades ago8,9,10, this method also relies on the emission/reception of ultrasounds, but with a radically different pattern. Indeed, instead of using a scanning focused beam, ultrafast imaging uses plane wave or diverging waves, which are able to insonify the field of view with a single emission. Following that single emission, the associated electronics is also able to receive and process the huge number of echoes originating from the whole field of view. At the end, an image can be reconstructed from a single emission/reception pattern11 (Figure 1, right panel). These unfocused emissions can have a low signal to noise ratio (SNR) due to the spread of the acoustical energy. This can be tackled by emitting several titled plane-waves (or diverging waves with different sources) and by adding the resulting images. This method is named “coherent compounding”12. Two major consequences arise. Firstly, the framerate only depends on the ultrasound time of flight and can reach typical values from 1 to 10 kHz. Secondly, this ensures the data continuity in both spatial and temporal dimensions, also referred to as spatiotemporal coherence. The conventional localization/quantification tradeoff is thus broken. This combination of a high framerate and spatiotemporal coherence has a tremendous impact on the ability to detect blood flows with ultrasound. As compared to conventional ultrasound, ultrafast ultrasound provides full characterization of the blood flow3. Practically, the user has access to the velocity time course in every pixel of the image, for the whole duration of the acquisition (typically ≈1 s), with a timescale given by the framerate (typically, a framerate of 5 kHz for a temporal resolution of 200 µs). This high framerate makes the method suitable for a wide range of application such as fast flow in moving organs like heart chambers13 or myocardium with the coronary micro-perfusion14. Furthermore, it has been shown that its spatiotemporal coherence strongly improves its ability to separate slow blood flow from background moving tissues, therefore increasing the sensitivity to micro-vascular flow15. This capacity gives access to the micro vasculature of the brain in both animals16 and humans17.
Hence, ultrafast ultrasound is well suited to image blood flow in a variety of situations. It is restricted to soft biological tissues and will be strongly affected by the presence of hard interfaces such as bones, or gas cavity such as the lung. The tuning of the physical parameters of the ultrasound sequence allows the study of both slow (down to 1 mm/s11,16) and fast flows (up to several m/s). A tradeoff exists between the spatial resolution and the depth of penetration. Typically, a resolution of 50 µm can be achieved at the cost of a penetration around 5 mm. Conversely, the penetration can be extended to 15-20 cm at the cost of a resolution of 1 mm. It is worth noting that most ultrafast scanners such as the one used in this article only provide 2D images.
Here, we propose a simple protocol to introduce the concept of Ultrafast Doppler imaging, using a programmable research ultrasound scanner and Doppler phantom mimicking a vessel (artery or vein) embedded in biological tissue.
1. Doppler phantom preparation setup (Figure 2A)
2. Ultrafast ultrasound scanner setup (Figure 2A)
3. Ultrasound sequence programming
4. Probe positioning and data acquisition
5. Image reconstruction (Figure 2B)
6. Clutter filtering (Figure 2C)
NOTE: For steps 6-7, see the Matlab script provided in the Supplementary Material.
7. Flow visualization and velocity measurements (Figure 2C)
The quality of the acquisition and the post-processing is firstly assessed by visual inspection. The shape of the canal must be clearly visible in the power Doppler image, and the tissue area must appear dark. If the power Doppler signal is not restricted to the canal, it can mean that either the clutter filter step went wrong (SVD threshold is too low), or the probe experienced a strong movement during the acquisition.
After visual inspection, the study of the spectrogram inside the canal can...
Several variations are possible around the main frame of this protocol.
Hardware concerns
If the user supplies its custom host computer, the motherboard and the computer’s case must have an available PCI express slot. The CPU must also have enough PCIe lanes to handle all the devices.
Probe selection
The ultrasound probe (also named transducer) is chosen according to the spatial resolution needed and to the geometr...
No conflict of interest
We would like to thank Shreya Shah for her proofreading and advice.
Name | Company | Catalog Number | Comments |
Blood-mimicking fluid | CIRS Inc, Norfolk, Virginia, USA | 069DTF | |
Doppler flow phantom | CIRS Inc, Norfolk, Virginia, USA | ATS523A | |
Matlab | MathWorks, Natick, Massachusetts, United States | ||
Peristaltic pump / Doppler flow pump | CIRS Inc, Norfolk, Virginia, USA | 769 | Include tubings and pulse dampener |
Transducer adpter | Verasonics, Kirkland, Washington, USA | UTA 408-GE | |
Ultrafast ultrasound research scanner | Verasonics, Kirkland, Washington, USA | Vantage 256 | |
Ultrasound probe/transducer | GE Healthcare | GE 9L-D |
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