While deep learning has catalyzed breakthroughs across numerous domains, its broader adoption in clinical settings is inhibited by the costly and time-intensive nature of data acquisition and annotation. To further facilitate medical machine learning, we present an ultrasound dataset of 10,223 Brightness-mode (B-mode) images consisting of sagittal slices of porcine spinal cords (N=25) before and after a contusion injury. We additionally benchmark the performance metrics of several state-of-the-art object detection algorithms to localize the site of injury and semantic segmentation models to label the anatomy for comparison and creation of task-specific architectures. Finally, we evaluate the zero-shot generalization capabilities of the segmentation models on human ultrasound spinal cord images to determine whether training on our porcine dataset is sufficient for accurately interpreting human data. Our results show that the YOLOv8 detection model outperforms all evaluated models for injury localization, achieving a mean Average Precision (mAP50-95) score of 0.606. Segmentation metrics indicate that the DeepLabv3 segmentation model achieves the highest accuracy on unseen porcine anatomy, with a Mean Dice score of 0.587, while SAMed achieves the highest Mean Dice score generalizing to human anatomy (0.445). To the best of our knowledge, this is the largest annotated dataset of spinal cord ultrasound images made publicly available to researchers and medical professionals, as well as the first public report of object detection and segmentation architectures to assess anatomical markers in the spinal cord for methodology development and clinical applications.

OBJECTIVE:In posterior vertebral column subtraction osteotomy (PVCSO), a section of a thoracic or lumbar vertebra and the adjacent disc are resected to shorten the spinal column, aiming to permanently alleviate tension on the spinal cord in patients with recurrent tethered cord syndrome (TCS). The effects of this procedure on the spinal cord are not well characterized. This study investigated morphometric changes in the cord following PVCSO and assessed associated clinical outcomes in patients with recurrent TCS. METHODS:A retrospective review of patients with recurrent TCS undergoing PVCSO with robotic assistance at the authors' tertiary care institution between 2019 and 2023 was performed. Clinical data were recorded from electronic medical records, and morphometric measurements, including T12-L2 sagittal height, intradural diameter, and the diameters, area, eccentricity, and positioning of the spinal cord, were collected from MRI. Spinal cord dimensions including anteroposterior and lateral diameters, area, eccentricity, positioning, and intradural diameter were compared before and after surgery. RESULTS:Six patients were included in this study. At 6-week follow-up, all patients had improvement on lower-extremity motor function examinations, 40% had improvement on lower-extremity sensory function examinations, and 83% had improved self-reported pain. Bladder and bowel incontinence were improved in 50% and 60%, respectively. PVCSO reduced the height of the spinal column by a mean of 18.1 ± 5.2 mm. PVCSO increased the mean spinal cord anteroposterior diameter by 0.8 ± 0.5 mm at T12 (p = 0.03) and the mean area by 0.4 ± 0.3 mm2 at T12 (p = 0.03). The mean eccentricity of the spinal cord decreased by 0.15 ± 0.15 at L1 (p = 0.05), indicating that the spinal cord became more circular after surgery. No major complications were reported, although 1 patient experienced atelectasis and pulmonary embolism postoperatively. CONCLUSIONS:This study provides novel insights into the morphometric changes induced by PVCSO and their correlation with clinical outcomes in patients with TCS. The procedure effectively increased spinal cord dimensions, alleviating tension and offering potential benefits in symptom relief. The study underscores the need for objective metrics to guide surgical decision-making and enhance the long-term success of PVCSO in the management of TCS.

Background Tension in the spinal cord is a trademark of tethered cord syndrome. Unfortunately, existing tests cannot quantify tension across the bulk of the cord, making the diagnostic evaluation of stretch ambiguous. A potential non-destructive metric for spinal cord tension is ultrasound-derived shear wave velocity (SWV). The velocity is sensitive to tissue elasticity and boundary conditions including strain. We use the term Ultrasound Tensography to describe the acoustic evaluation of tension with SWV. Methods Our solution Tethered cord Assessment with Ultrasound Tensography (TAUT) was utilized in three sub-studies: finite element simulations, a cadaveric benchtop validation, and a neurosurgical case series. The simulation computed SWV for given tensile forces. The cadaveric model with induced tension validated the SWV-tension relationship. Lastly, SWV was measured intraoperatively in patients diagnosed with tethered cords who underwent treatment (spinal column shortening). The surgery alleviates tension by decreasing the vertebral column length. Results Here we observe a strong linear relationship between tension and squared SWV across the preclinical sub-studies. Higher tension induces faster shear waves in the simulation ( R 2 = 0.984) and cadaveric ( R 2 = 0.951) models. The SWV decreases in all neurosurgical procedures ( p < 0.001). Moreover, TAUT has a c-statistic of 0.962 (0.92-1.00), detecting all tethered cords. Conclusions This study presents a physical, clinical metric of spinal cord tension. Strong agreement among computational, cadaveric, and clinical studies demonstrates the utility of ultrasound-induced SWV for quantitative intraoperative feedback. This technology is positioned to enhance tethered cord diagnosis, treatment, and postoperative monitoring as it differentiates stretched from healthy cords.

Focused ultrasound (FUS) is an increasingly prevalent therapeutic tool used in medicine to treat patients non-invasively, cost-effectively, and without any ionizing radiation. Depending on the clinical application, therapeutic ultrasound can be used for tissue ablation, physiotherapy, lithotripsy, and drug delivery, among other therapies. For the treatment to be effective, it is important to understand the acoustic pressure profile in the tissue to ensure the focal point intensity is sufficient to achieve therapy without damaging the tissue. In spinal cord injury (SCI), where FUS is hypothesized to promote blood flow to the site of injury to facilitate healing, it can be difficult to predict the expected pressure wavefields in real-time. This challenge is due to the complex structure of the spinal cord and variations in acoustic properties within the field of view, leading to distortion of the ultrasound wave front. To address this issue, we present an automated approach to visualize beam propagation and acoustic pressure distribution to aid with placement of the FUS probe. By simulating propagation of the ultrasound beams through patient-specific Digital Imaging and Communications in Medicine (DICOM) images, therapy can be optimized to minimize tissue damage while providing adequate focusing and focal intensity. In this study, porcine spinal cord images acquired after acute SCI are used to determine optimal FUS transducer placement.

Tissue elasticity remains an essential biomarker of health and is indicative of irregularities such as tumors or infection. The timely detection of such abnormalities is crucial for the prevention of disease progression and complications that arise from late-stage illnesses. However, at both the bedside and the operating table, there is a distinct lack of tactile feedback for deep-seated tissue. As surgical techniques advance toward remote or minimally invasive options to reduce infection risk and hasten healing time, surgeons lose the ability to manually palpate tissue. Furthermore, palpation of deep structures results in decreased accuracy, with the additional barrier of needing years of experience for adequate confidence of diagnoses. This review delves into the current modalities used to fulfill the clinical need of quantifying physical touch. It covers research efforts involving tactile sensing for remote or minimally invasive surgeries, as well as the potential of ultrasound elastography to further this field with non-invasive real-time imaging of the organ's biomechanical properties. Elastography monitors tissue response to acoustic or mechanical energy and reconstructs an image representative of the elastic profile in the region of interest. This intuitive visualization of tissue elasticity surpasses the tactile information provided by sensors currently used to augment or supplement manual palpation. Focusing on common ultrasound elastography modalities, we evaluate various sensing mechanisms used for measuring tactile information and describe their emerging use in clinical settings where palpation is insufficient or restricted. With the ongoing advancements in ultrasound technology, particularly the emergence of micromachined ultrasound transducers, these devices hold great potential in facilitating early detection of tissue abnormalities and providing an objective measure of patient health.

Abstract Background Tension in the spinal cord is a trademark of tethered cord syndrome. Unfortunately, existing tests cannot quantify tension across the bulk of the cord, making the diagnostic evaluation of stretch ambiguous. A potential non-destructive metric for spinal cord tension is ultrasound-derived shear wave velocity (SWV). The velocity is sensitive to tissue elasticity and boundary conditions including strain. We use the term Ultrasound Tensography to describe the acoustic evaluation of tension with SWV. Methods Our solution “Tethered cord Assessment with Ultrasound Tensography (TAUT)” was utilized in three sub-studies: finite element simulations, a cadaveric benchtop validation, and a neurosurgical case series. The simulation computed SWV for given tensile forces. The induced tension cadaveric model validated the SWV-tension relationship. Lastly, SWV was measured intraoperatively in patients diagnosed with tethered cord who underwent surgical treatment (spinal column shortening). The surgery alleviates tension by decreasing the vertebral column length. Results Here we observe a strong linear relationship between tension and squared SWV across the preclinical sub-studies. Higher tension induces faster shear waves in the simulation (R2 = 0.984) and cadaveric (R2 = 0.951) models. The SWV decreases in all neurosurgical procedures (p<0.001). Moreover, TAUT has a c-statistic of 0.962 (0.92-1.00), detecting all tethered cords. Conclusions This study presents the first clinical metric of spinal cord tension. Strong agreement among computational, cadaveric, and clinical studies demonstrates the utility of ultrasound-induced SWV for quantitative intraoperative feedback. This technology is positioned to enhance tethered cord diagnosis, treatment, and post-operative monitoring as it differentiates stretched from healthy cords.

Ultrasound holds promise for use in spinal cord injury cases for both diagnostic and therapeutic purposes. Focused ultrasound applications demand an added threshold of study to ensure the safety and efficacy of the therapy. For optimal treatment outcomes, it is crucial to understand whether relevant structures are being targeting with sufficient energy without damaging neighboring tissue and vasculature. However, it is difficult to predict the expected displacement and pressure profile of the ultrasound wavefront due to challenges with visualizing an acoustic beam in real-time and complex patient-specific anatomy. This challenge is particularly prominent in anatomies with varying medium acoustic properties that cause reflection and distortion of the signal, which is inherent to the composition of the spinal cord and is exacerbated by the formations of injury-induced hematomas. Incorrect placement of focused ultrasound transducers can be detrimental to patient health, specifically if therapeutic ultrasound is used at higher intensities, as the beam propagation can target healthy tissue and important structures that could lead to tissue damage and death. We study how computational tools can be leveraged to aid placement of the transducer using an ultrasound simulation software, Wave 3000 Plus, that allows for the visualization of ultrasound propagation through anatomical structures. By simulating the propagation of ultrasound beams through patient-specific Digital Imaging and Communications in Medicine (DICOM) images, we study computational approaches to determine the optimal placement of devices. In this study, we use in vivo porcine spinal cord images following spinal cord injury (as an example medical use case) to determine if the injury site is being targeted appropriately and to visualize the distribution of pressure throughout the simulation. We demonstrate that Wave 3000 Plus is a viable approach for visualizing ultrasound propagation through patient-specific anatomies.

Approximately 900,000 spinal cord injuries (SCI) occur each year. Understanding the severity and progression of the injury can help tailor the treatment plan to optimize a patient’s prognosis. One way to determine the progression of a neurological injury is to monitor the blood flow. While software exists for quantifying renal tissue perfusion with Doppler ultrasound, we developed an algorithm optimized to quantify perfusion in spinal cord microvasculature with multiple ultrasound imaging modalities. The objective of this study was to demonstrate spinal cord microvascular quantification methods using non-contrast ultrasound images. Following a T4-T6 laminectomy, ultrasound videos were captured of in vivo porcine spinal cords using color Doppler (CDI), advanced dynamic flow (ADF), and superb microvascular imaging (SMI) modalities. A MATLAB algorithm was developed to import ultrasound videos, extract the velocity map, and quantify the microvasculature blood flow as a function of time by averaging the velocity map in a region of interest. Using the velocity-time curve (VTC), local stroke volume (LSV) and local vascular output (LVO) were calculated. Our algorithm detected slow-flow (< 0.3 cm/s) changes indicative of cardiac cycles in each ultrasound modality for sub-millimeter diameter vessels. Each cardiac cycle from the VTC was extracted to calculate LSV and LVO. The mean ± 1 standard deviation LVO for CDI, ADF, and SMI were 0.23±0.008 mL/min, 0.28±0.003 mL/min, and 0.18±0.004 mL/min, respectively. Calculating these local perfusion metrics and tracking local perfusion after SCI may supplement current treatment by reducing the dependence on global measures of blood flow (e.g., mean arterial pressure).

INTRODUCTION: Tethered cord syndrome (TCS) has been traditionally treated with detethering surgery.1,2 Posterior vertebral column subtraction osteotomy (PVCSO) is a novel surgery that can treat TCS while avoiding complications of detethering surgery, such as retethering or cerebrospinal fluid leaks.2-6 The pathophysiology of PVCSO is not completely understood. However, the hypothesis is that shortening the vertebral column decreases the spinal cord (SC) tension thereby improving neurological function. Intraoperative ultrasound elastography (USE) is well suited to elucidate the changes that can occur in SC parenchyma secondary to PVCSO.7,8 Through different modalities, such as superb microvascular imaging (SMI) and elastography (USE), US can analyze changes in blood flow and tissue tension.9 METHODS: Three patients with recurrent TCS underwent PVCSO and intraoperative USE to analyze the tension of the SC at 3 timepoints: pre-, mid- and post-shortening. RESULTS: All three patients (two female, one male) had undergone multiple detethering surgeries. Prior to shortening, the median SC elasticity of all patients was 9.8 kPa (IQR 9.7-12.4 kPa). After complete compression of the rods, the SC elasticity decreased by 17.4% to 7.7 kPa (6.5-9.7 kPa). All patients had significant improvement in sensation and reduction in pain at their 2 month follow up appointment. CONCLUSIONS: Here we report the first use of USE to demonstrate decreased SC tension after PVCSO for TCS. All patients reported improved symptoms and USE demonstrated decreased SC tension after shortening. USE is a promising tool for evaluating tissue elasticity and blood flow in spinal pathologies, including TCS. This technology serves as an intraoperative confirmation that SC tension decreases after PVCSO. Patients and neurosurgeons both benefit from an intraoperative objective tool to demonstrate reduced SC tension.

Objective: Here we investigate the ability of low-intensity ultrasound (LIUS) applied to the spinal cord to modulate the transmission of motor signals. Methods: Male adult Sprague-Dawley rats (n = 10, 250–300 g, 15 weeks old) were used in this study. Anesthesia was initially induced with 2% isoflurane carried by oxygen at 4 L/min via a nose cone. Cranial, upper extremity, and lower extremity electrodes were placed. A thoracic laminectomy was performed to expose the spinal cord at the T11 and T12 vertebral levels. A LIUS transducer was coupled to the exposed spinal cord, and motor evoked potentials (MEPs) were acquired each minute for either 5- or 10-minutes of sonication. Following the sonication period, the ultrasound was turned off and post-sonication MEPs were acquired for an additional 5 minutes. Results: Hindlimb MEP amplitude significantly decreased during sonication in both the 5- (p < 0.001) and 10-min (p = 0.004) cohorts with a corresponding gradual recovery to baseline. Forelimb MEP amplitude did not demonstrate any statistically significant changes during sonication in either the 5- (p = 0.46) or 10-min (p = 0.80) trials. Conclusion: LIUS applied to the spinal cord suppresses MEP signals caudal to the site of sonication, with recovery of MEPs to baseline after sonication. Significance: LIUS can suppress motor signals in the spinal cord and may be useful in treating movement disorders driven by excessive excitation of spinal neurons.

OBJECTIVES/GOALS: This poster shares a case study on how a group at The Johns Hopkins University formed a translational lab missioned to reinvent currently existing treatments for acute spinal cord injuries, implanting in humans within a five-year window. The poster showcases how a project funded by the Defense Advanced Research Projects Agency has been implemented. METHODS/STUDY POPULATION: The translational team; Holistic Electrical; ultrasonic and Physiological Interventions Unburdening those with Spinal cord injury• (HEPIUS) Lab is composed of many parts as listed below: neurosurgeons; engineers; radiologists; public health specialists; statisticians; patient advocates; ethicists; sonographers; researchers; academic collaborators; and specialized industry partners. Sometimes physically separated; the team has videoconferencing carts across locations to stay connected at every step in the process. The lab facilities were organized with several key facets in mind: research and development (R&D); prototyping; fabrication; verification; and validation (V&V); animal model testing; cadaveric testing accessibility; mock operating room for simulations; and collaboration hubs. RESULTS/ANTICIPATED RESULTS: Due to communications with the US Food and Drug Administration (FDA), DARPA, patient advocates, ethicists, internal review boards, and other bodies, the team has a clear path towards clinical translation. The team has the following stages in progress or scheduled: manufacturing devices, benchtop testing, rat and pig models, biocompatibility testing, cadaveric testing, and clinical use. The lab space was designed to achieve these core functions. For rapid, in-house manufacturing, the lab has unique capabilities including 3D metal printing. For experiments, industry collaborations and equipment acquisitions enable the highest quality research. These technologies are assembled into diagnostic, therapeutic, testing, and manufacturing hubs to drive real change in the lives of many; the patient comes first. DISCUSSION/SIGNIFICANCE: This laboratory, team, and system of operation is aimed to enable novel practices for the clinical translation of spinal cord medical solutions. For researchers interested in launching their own translational work, this poster may serve as a reference, example, and inspiration for similar hopeful university-centered hubs.

Spinal cord injury (SCI) is a devastating disease with limited effective treatment options. Animal paradigms are vital for understanding the pathogenesis of SCI and testing potential therapeutics. The porcine model of SCI is increasingly favored because of its greater similarity to humans. However, its adoption is limited by the complexities of care and range of testing parameters. Researchers need to consider swine selection, injury method, post-operative care, rehabilitation, behavioral outcomes, and histology metrics. Therefore, we systematically reviewed full-text English-language articles to evaluate study characteristics used in developing a porcine model and summarize the interventions that have been tested using this paradigm. A total of 63 studies were included, with 33 examining SCI pathogenesis and 30 testing interventions. Studies had an average sample size of 15 pigs with an average weight of 26 kg, and most used female swine with injury to the thoracic cord. Injury was most commonly induced by weight drop with compression. The porcine model is amenable to testing various interventions, including mean arterial pressure augmentation (n = 7), electrical stimulation (n = 6), stem cell therapy (n = 5), hypothermia (n = 2), biomaterials (n = 2), gene therapy (n = 2), steroids (n = 1), and nanoparticles (n = 1). It is also notable for its clinical translatability and is emerging as a valuable pre-clinical study tool. This systematic review can serve as a guideline for researchers implementing and testing the porcine SCI model.

Elastography is an imaging technology capable of measuring tissue stiffness and consistency. The technology has achieved widespread use in the workup and management of diseases of the liver, breast, thyroid, and prostate. Although elastography is increasingly being applied in neurosurgery, it has not yet achieved widespread adoption and many clinicians remain unfamiliar with the technology. Therefore, we sought to summarize the range of applications and elastography modalities available for neurosurgery, report its effectiveness in comparison with conventional imaging methods, and offer recommendations. All full-text English-language manuscripts on the use of elastography for neurosurgical procedures were screened using the PubMed/MEDLINE, Embase, Cochrane Library, Scopus, and Web of Science databases. Thirty-two studies were included with 990 patients, including 21 studies on intracranial tumors, 5 on hydrocephalus, 4 on epilepsy, 1 on spinal cord compression, and 1 on adolescent scoliosis. Twenty studies used ultrasound elastography (USE) whereas 12 used magnetic resonance elastography (MRE). MRE studies were mostly used in the preoperative setting for assessment of lesion stiffness, tumor-brain adherence, diagnostic workup, and operative planning. USE studies were performed intraoperatively to guide resection of lesions, determine residual microscopic abnormalities, assess the tumor-brain interface, and study mechanical properties of tumors. Elastography can assist with resection of brain tissue, detection of microscopic lesions, and workup of hydrocephalus, among other applications under investigation. Its sensitivity often exceeds that of conventional MRI and ultrasound for identifying abnormal tissue and lesion margins.

Imaging of spinal cord microvasculature holds great potential in directing critical care management of spinal cord injury (SCI). Traditionally, contrast agents are preferred for imaging of the spinal cord vasculature, which is disadvantageous for long-term monitoring of injury. Here, we present FlowMorph, an algorithm that uses mathematical morphology techniques to segment non-contrast Doppler-based videos of rat spinal cord. Using the segmentation, it measures single-vessel parameters such as flow velocity, rate, and radius, with visible cardiac cycles in individual vessels showcasing the spatiotemporal resolution. The segmentation outlines vessels well with little extraneous labeling, and outlines are smooth through time. Radius measurements of perforating vessels are similar to what is seen in the literature through other methods. Verification of the algorithm through comparison to manual measurement and in vitro microphantom standards highlights points of future improvement. This method will be vital for future work studying the vascular effects of SCI and can be adopted to other species as well.

OBJECTIVES/GOALS: The first aim was to construct a controlled and high resolution FUS water tank characterization system with 1 micron step-sizes. The second aim was to create two unique standardized protocols for mapping the generated acoustic field from FUS transducers; protocol one maps the full 3D field while protocol two rapidly detects changes to the original plot. METHODS/STUDY POPULATION: To accomplish aim one, the focused ultrasound mapping platform was constructed with a water conditioning unit for water degassing and temperature control, a three-axis stage with 1 micron step-size capabilities, and a data plotting software. To measure the outcomes of aim one, the water temperature was monitored, and axis step sizes were measured through ten independent axis translation recordings. To accomplish aim two, FUS acquisitions were executed at different resolutions. For FUS localization at the cellular level, a 1-5 micron step size is required. Once the initial scan was performed, duplicate scans were executed to detect inherent perturbations or errors in the system. Once calculated, the best methods of detecting true changes to FUS signals are proposed. RESULTS/ANTICIPATED RESULTS: The FUS characterization system maintained water temperature and performed 1 micron step-sizes. While pre-existing platforms have demonstrated a resolution of one thousand recordings per cubic millimeter, the proposed system (time and computing power willing) can record one billion recordings per cubic millimeter. In practice, a resolution of 20 micron was sufficient for non-cellular level FUS characterizations. Successive 2D scans were reliably stacked to form a 3D rendering of the generated acoustic field with the average focal point intensity yielding a 1% coefficient of variation between identical scans. This inherent variation can be used as the threshold of significance for true change detection; to rapidly detect changes to the FUS signal, sampling can be performed at regions of high baseline values. DISCUSSION/SIGNIFICANCE: Focused ultrasound medical devices are gaining popularity for treatments including tumor ablation, neuromodulation, and drug delivery; however, the field lacks a standardized method to characterize these FUS transducers. The presented platform and protocols enable a rigorous and high quality translation through verification and validation.

Focused ultrasound (FUS) is an emerging technique with the potential to revolutionize traditional treatment methods in the fields of oncology and neurosurgery. Recently, FUS treatments have shown potential for altering neural activity in the spinal cord, with the intent to alleviate pain. Preliminary animal studies using FUS have demonstrated the need for transducer accessories that can simplify the implementation of the transducer in the clinic. The coupling cone that was supplied with the transducer was designed for larger target tissues. Thus, surgeons have expressed a desire to adapt the cone design to be easier to use for smaller targets, such as the spinal cord. Here, we developed 3D printed cones, with smaller aperture sizes, for FUS transducers to assist surgeons in localizing the focal point of the transducers in a faster, and more intuitive manner. The cones were designed to not alter the original focal region of the transducers. This was experimentally confirmed by measuring the size of the focal region for the transducer with the new cones and comparing this data to measurements provided by the manufacturer. The new coupling cones will make the FUS transducers more ergonomic for use in stimulating the spinal cord in an animal model.

Glioblastoma Multiforme (GBM) is a malignant brain cancer with low overall survival. Therefore, researchers are looking to augment its current therapeutic regimen, which includes surgical tumor resection, chemotherapy and radiation. A promising treatment modality, focused ultrasound, has been used as a non-invasive treatment for GBM through multiple approaches such as thermal ablation, immunomodulation, and blood brain barrier disruption. In order to develop these treatments for clinical trials, testing in animal models needs to be performed to investigate the efficacy of the treatment in complex biological environments, as well as to evaluate any side-effects. The more biologically relevant the animal model is to human anatomy, the more applicable the results will be for translation to clinical trials. Here, we report a human GBM rat model, which utilizes an IDH-wildtype, EGFRvIII mutant patient-derived xenograft in athymic rats. The in vivo tumor growth rate was assessed over a period of 20 days to evaluate reproducibility and to develop the model for future testing of FUS in the treatment of GBM.

Focused ultrasound (FUS) is becoming widely researched for medical therapies due to its high penetration depth, spatial resolution, and affordability. Applications of FUS range from high intensity focused ultrasound (HIFU) for the ablation of cancerous tumors to low intensity focused ultrasound (LIFU) for the treatment of neurological conditions like essential tremors. A key step in developing these treatments and their corresponding FUS devices is characterizing the emitted ultrasound from the proposed transducer. However, a bottleneck exists at this verification and validation stage; current characterization techniques lack the robustness of reliably recording below a 5μm resolution. This level of accuracy is needed to adequately design devices which can target cells like astrocytes or other desired target tissues at this scale. Our Acoustic Measurement Platform for Localizing and Implementing Therapeutic Ultrasound Devices and Equipment (AMPLITUDE) is a solution which enables engineers, scientists, and clinicians to confidently characterize their equipment in a benchtop setting. It achieves this resolution by utilizing an all-in-one water conditioning unit, linear stepper motors with a theoretical step size of 1 μm and a 1% standard deviation on repetitive experiments, as well as signal processing techniques. This system can be used throughout the product timeline including prototyping, verifying efficacy, FDA testing, and routine check-ups during clinical use.