A 1.11 mm2 IVUS SoC with ±50°-Range Plane Wave Transmit Beamforming at 40 MHz.

Intravascular ultrasound (IVUS) imaging catheters are significant tools for cardiovascular interventions, and their use can be expanded by realizing IVUS imaging guidewires and microcatheters. The miniaturization of these devices creates challenges in SNR due to the need for higher frequencies to provide adequate resolution. An integrated IVUS system with transmit beamforming can mitigate these limitations. This work presents the first practical highly integrated system-on-a-chip (SoC) with plane wave transmit beamforming at 40 MHz for IVUS on guidewire or microcatheters. The front-end circuitry has a 20-channel ultrasound transmitter (Tx) and receiver (Rx) array interfaced with a capacitive micromachined ultrasound transducer (CMUT) array. During each firing, all 20 Tx are excited with the same analog delay with respect to each other, which can be continuously adjusted between ~0 and 10 ns in two directions, generating a steerable plane wave in a range of ±/-50° for a phased array at 40 MHz. The unit delays are generated via a voltage-controlled delay line (VCDL), which only needs two external controls, one tuning the unit delay and the other determining the steering direction. The SoC is fabricated using a 180-nm high-voltage (HV) CMOS process and features a slender active area of 0.3 mm × 3.7 mm. The proposed SoC consumes 31.3 mW during the receiving mode. The beamformer's functionality and the SoC's overall performance were validated through acoustic characterization and imaging experiments.

Spatially targeted brain cancer immunotherapy with closed-loop controlled focused ultrasound and immune checkpoint blockade.

Despite the challenges in treating glioblastomas (GBMs) with immune adjuvants, increasing evidence suggests that targeting the immune cells within the tumor microenvironment (TME) can lead to improved responses. Here, we present a closed-loop controlled, microbubble-enhanced focused ultrasound (MB-FUS) system and test its abilities to safely and effectively treat GBMs using immune checkpoint blockade. The proposed system can fine-tune the exposure settings to promote MB acoustic emission-dependent expression of the proinflammatory marker ICAM-1 and delivery of anti-PD1 in a mouse model of GBM. In addition to enhanced interaction of proinflammatory macrophages within the PD1-expressing TME and significant improvement in survival (P < 0.05), the combined treatment induced long-lived memory T cell formation within the brain that supported tumor rejection in rechallenge experiments. Collectively, our findings demonstrate the ability of MB-FUS to augment the therapeutic impact of immune checkpoint blockade in GBMs and reinforce the notion of spatially tumor-targeted (loco-regional) brain cancer immunotherapy.

Integrated Hybrid Sub-Aperture Beamforming and Time-Division Multiplexing for Massive Readout in Ultrasound Imaging.

This paper demonstrates hybrid sub-aperture beamforming (SAB) with time-division multiplexing (TDM) for massive interconnect reduction in ultrasound imaging systems. A single-chip front-end system prototype has been fabricated in 180-nm HV BCD technology that combines 5×1 SAB with 8×1 TDM to efficiently reduce the number of receive signal interconnects by a factor of 40. The system includes on-chip high-voltage (HV) pulsers capable of generating unipolar pulses up to 70 V in transmit (TX) mode. The receiver (RX) chain consists of a T/R switch, a variable-gain low-noise amplifier (VG-LNA) with 4-step gain control (15-32 dB) for time-gain compensation followed by a programmable switched-capacitor analog delay-and-sum beamformer. The proof-of-concept prototype operates at a 200-MHz clock frequency and the SAB provides 32-step fine delays with a maximum delay of 310 ns corresponding to better than λ/20 delay quantization at 5 MHz. With these specifications, the SAB is capable of beam steering from 0 ° to 45 ° for a 5-element subarray with 150-micron pitch ( λ/2), providing a near-ideal phased array imaging performance. The sub-aperture beamformer is followed by the TDM system where each of the 8 channels is sampled at a rate of 25 MS/s after an anti-aliasing bandpass filter. The full functionality of the prototype chip is validated through electrical and acoustic measurements on a 1-D capacitive micromachined ultrasonic transducer (CMUT) array designed for intracardiac echocardiography (ICE).

An Adaptive Element-Level Impedance-Matched ASIC With Improved Acoustic Reflectivity for Medical Ultrasound Imaging.

This paper presents an active impedance matching scheme that tries to optimize electrical power transfer and acoustic reflectivity in ultrasound transducers. Leveraging negative capacitance-based impedance matching would potentially improve the bandwidth and electrical power transfer while minimizing acoustic reflection of transducer elements and improve uniformity while reducing acoustic crosstalk of transducer arrays. A 16-element transceiver front-end is designed which employs an element-level active capacitive impedance cancellation scheme using an element-level negative impedance converter. The ASIC fabricated in 180-nm HVBCD technology provides high-voltage pulses up to 60 V consuming 3.6 mW and occupying 2.5 mm2. The front-end ASIC is used with a 1-D capacitive micromachined ultrasonic transducer (CMUT) array and its acoustical reflectivity reduction and imaging capabilities have successfully been demonstrated through pulse-echo measurements and acoustic imaging experiments.

Towards controlled drug delivery in brain tumors with microbubble-enhanced focused ultrasound.

Brain tumors are particularly challenging malignancies, due to their location in a structurally and functionally distinct part of the human body - the central nervous system (CNS). The CNS is separated and protected by a unique system of brain and blood vessel cells which together prevent most bloodborne therapeutics from entering the brain tumor microenvironment (TME). Recently, great strides have been made through microbubble (MB) ultrasound contrast agents in conjunction with ultrasound energy to locally increase the permeability of brain vessels and modulate the brain TME. As we elaborate in this review, this physical method can effectively deliver a wide range of anticancer agents, including chemotherapeutics, antibodies, and nanoparticle drug conjugates across a range of preclinical brain tumors, including high grade glioma (glioblastoma), diffuse intrinsic pontine gliomas, and brain metastasis. Moreover, recent evidence suggests that this technology can promote the effective delivery of novel immunotherapeutic agents, including immune check-point inhibitors and chimeric antigen receptor T cells, among others. With early clinical studies demonstrating safety, and several Phase I/II trials testing the preclinical findings underway, this technology is making firm steps towards shaping the future treatments of primary and metastatic brain cancer. By elaborating on its key components, including ultrasound systems and MB technology, along with methods for closed-loop spatial and temporal control of MB activity, we highlight how this technology can be tuned to enable new, personalized treatment strategies for primary brain malignancies and brain metastases.

Analysis of Negative Capacitance-Based Broadband Impedance Matching for CMUTs.

Tight integration of capacitive micromachined ultrasonic transducer (CMUT) arrays with integrated circuits can make active impedance matching feasible for practical imaging devices. In this article, negative capacitance-based impedance matching for CMUTs is investigated. Simple equivalent circuit model-based calculations show the potential of negative capacitance matching for improving the bandwidth along with electrical power transfer and acoustic reflectivity, but the model has limitations especially for acoustic reflectivity evaluation. For more realistic results, an experimentally validated CMUT array model is applied to a small 1-D CMUT array operating in the 5-15 MHz range. The results highlight the difference between electrical power transfer and acoustic reflectivity as well as the tradeoffs in signal-to-noise ratio (SNR). According to the results, ideal negative capacitance termination matched to the CMUT capacitance provides the broadest bandwidth and highest SNR if acoustic or electrical reflections are of no concern. On the other hand, negative capacitance and resistance matching to minimize acoustic reflectivity provides both lower reflection and closer to ideal SNR as compared with electrical power matching. It is observed that acoustic matching also reduces acoustic crosstalk and improves array uniformity. While several challenges for integrated circuit implementation are present, negative capacitance-based impedance matching can be a viable broadband active impedance matching method for CMUTs operating in conventional and collapsed mode as well as other ultrasound transducers with mainly capacitive impedance.

Real-time device tracking under MRI using an acousto-optic active marker.

PURPOSE: This work aims to demonstrate the use of an "active" acousto-optic marker with enhanced visibility and reduced radiofrequency (RF) -induced heating for interventional MRI. METHODS: The acousto-optic marker was fabricated using bulk piezoelectric crystal and π-phase shifted fiber Bragg grating (FBGs) and coupled to a distal receiver coil on an 8F catheter. The received MR signal is transmitted over an optical fiber to mitigate RF-induced heating. A photodetector converts the optical signal into electrical signal, which is used as the input signal to the MRI receiver plug. Acousto-optic markers were characterized in phantom studies. RF-induced heating risk was evaluated according to ASTM 2182 standard. In vivo real-time tracking capability was tested in an animal model under a 0.55T scanner. RESULTS: Signal-to-noise ratio (SNR) levels suitable for real-time tracking were obtained by using high sensitivity FBG and piezoelectric transducer with resonance matched to Larmor frequency. Single and multiple marker coils integrated to 8F catheters were readout for position and orientation tracking by a single acousto-optic sensor. RF-induced heating was significantly reduced compared to a coax cable connected reference marker. Real-time distal tip tracking of an active device was demonstrated in an animal model with a standard real-time cardiac MR sequence. CONCLUSION: Acousto-optic markers provide sufficient SNR with a simple structure for real-time device tracking. RF-induced heating is significantly reduced compared to conventional active markers. Also, multiple RF receiver coils connected on an acousto-optic modulator can be used on a single catheter for determining catheter orientation and shape.

A Power-Efficient Bridge Readout Circuit for Implantable, Wearable, and IoT Applications.

A power-efficient bridge-to-digital sensing interface is proposed, which also offers immunity against power supply noise. The interface utilizes duty-cycling to reduce the static power consumption of resistive bridge sensors, which are commonly used in implantable, wearable, and internet of things (IoT) applications, such as intracranial pressure (ICP) sensing and blood pressure (BP) monitoring. The proposed interface uses a revised version of the pseudo-pseudo differential (PPD) topology with the ping-pong technique to reduce the complexity of traditional fully-differential counterparts. A proof-of-concept prototype has been fabricated in 0.35-µm CMOS and occupies an active area of 0.48 mm2. It achieves 9.13 effective number of bits (ENOB) at 3.72 kHz sampling rate and improvement of more than 50 dB in the power supply rejection ratio (PSRR) by employing the ping-pong technique. It reduces the power consumption of a 5-kΩ Wheatstone bridge by 99.6% compared to a traditional interface, down to 2.53 µw at 1.8 V supply. The functionality of the system has also been demonstrated in an experimental setup in conjunction with an embedded resistive bridge pressure sensor.

Sensitivity and phase response of FBG based acousto-optic sensors for real-time MRI applications.

Fiber Bragg grating (FBG) based sensors have recently been introduced to the field of magnetic resonance imaging (MRI). Real-time MRI applications demand highly amplitude and phase sensitive MRI compatible sensors. Thus, a model and detailed analysis of FBG based ultrasound detection are required for designing better performing sensors. A hybrid FBG model incorporating numerical and FEA methods was developed and used for sensitivity and linearity analysis. The transfer matrix method was used for the modeling of optical modulation whereas FEA was used for pressure field calculations within the grating. The model was verified through reflection spectrum and acoustic pressure sensitivity testing of two π-phase shifted FBGs in a side slope read-out configuration. The sensitivity curves with respect to the operation point on the side slope was characterized in terms of amplitude and phase, and nonlinearity of the phase response has been quantified. Lastly, the impact of phase linearity of the FBG based acousto-optic sensor was tested under MRI when the sensor was used as a position marker and an analog phase shifter based solution was demonstrated.

Highly Integrated Guidewire Ultrasound Imaging System-on-a-Chip.

In this article, we present a highly integrated guidewire ultrasound (US) imaging system-on-a-chip (GUISoC) for vascular imaging. The SoC consists of a 16-channel US transmitter (Tx) and receiver (Rx) electronics, on-chip power management IC (PMIC), and quadrature sampler. Using a synthetic aperture imaging algorithm, a Tx/Rx pair, connected to capacitive micromachined ultrasound transducers (CMUTs), can be activated at any time. The Tx generates acoustic waves by driving the CMUT, while the Rx picks up the echo signal and amplify it to be delivered through an interconnect that is driven by a buffer. On-chip logic controls the pulsers that generate the high-voltage (HV)-pulse for Tx. An on-chip PMIC provides 1.8-, 5-, 39-, and 44-V supplies and a clock signal from the two interconnects besides GND. A quadrature sampler down-converts the Rx echo signal to baseband, reducing its bandwidth requirement for the output interconnect. The system design, including transimpedance amplifier (TIA) optimization, based on the equivalent circuit of a specific CMUT is presented. The SoC was fabricated by a 0.18-µm HV CMOS process, occupying 1.5-mm2 active area and consuming 25.2 and 44 mW from 1.8 to 44 V supplies, respectively. The US Tx and Rx show bandwidths of 32-42 and 32.7-37.5 MHz, respectively. The input-referred noise of the system was measured as 9.66 nA in band with 2-m-long 52 American Wire Gauge (AWG) wire interconnects. The functionality of the GUISoC was verified in vitro by imaging wire targets.

An Impulse Radio PWM-Based Wireless Data Acquisition Sensor Interface.

A sensor interface circuit based on impulse radio pulse width modulation (IR-PWM) is presented for low power and high throughput wireless data acquisition systems (wDAQ) with extreme size and power constraints. Two triple-slope analog-to-time converters (ATC) convert two analog signals, each up to 5 MHz in bandwidth, into PWM signals, and an impulse radio (IR) transmitted (Tx) with an all-digital power amplifier (PA) combines them while preserving the timing information by transmitting impulses at the PWM rising and falling edges. On the receiver (Rx) side, an RF-LNA followed by an envelope detector recovers the incoming impulses, and a T-flipflop reverts the impulse sequence back to PWM to be digitized by a time-to-digital converter (TDC). Detailed analysis and design guideline on ATC was introduced, and a proof-of-concept prototype was fabricated for a capacitive micromachined ultrasound transducer (CMUT) imaging system in a 0.18-µm HV CMOS process, occupying 0.18 mm2 active area and consuming 3.94 mW from a 1.8 V supply. The proposed TDC in this prototype yielded 7-bit resolution, while the entire wDAQ achieved 5.8 effective number of bits (ENOB) at 2 × 10 MS/s.

An Analysis Method for Capacitive Micromachined Ultrasound Transducer (CMUT) Energy Conversion during Large Signal Operation.

With Capacitive Micromachined Ultrasound Transducers (CMUTs) increasingly being used for high intensity, large signal ultrasound applications and several drive methods being proposed, the efficiency of these devices in this operation regime have not been quantitatively evaluated. Since well-known frequency and capacitance-based coupling coefficients definitions are not valid for large signal, nonlinear operation, an energy-based definition should be used. In this paper, an expression for mechanical energy in a CMUT is obtained based on the assumption that CMUT is a linear time varying capacitor in all regimes of operation. This expression is evaluated by the help of an experimentally verified nonlinear CMUT model to define an energy conversion ratio (ECR) which can be considered as a coupling coefficient valid for all regimes of operation. This parameter is validated in the small signal regime and then used to evaluate CMUT performance with various large drive signals. The quantitative modeling results show that CMUTs do not need DC bias to achieve high efficiency large signal transduction: AC only signals at half the operation frequency with amplitudes beyond the collapse voltage can provide efficiencies (ECR) above 0.9 with harmonic content below -25 dB. Based on these results, ECR variation with membrane geometry and parasitic capacitance are given as examples for device optimization. The overall modeling approach is also qualitatively validated by experiments.

Supply-Inverted Bipolar Pulser and Tx/Rx Switch for CMUTs Above the Process Limit for High Pressure Pulse Generation.

A combined supply-inverted bipolar pulser and a Tx/Rx switch is proposed to drive capacitive micromachined ultrasonic transducers (CMUTs). The supply-inverted bipolar pulser adopts a bootstrap circuit combined with stacked transistors, which guarantees high voltage (HV) operation above the process limit without lowering device reliability. This circuit generates an output signal with a peak-to-peak voltage that is almost twice the supply level. It generates a bipolar pulse with only positive supply voltages. The Tx/Rx switch adopts a diode-bridge structure with the protection scheme dedicated to this proposed pulser. A proof- of-concept ASIC prototype has been implemented in 0.18-µm HV CMOS/DMOS technology with 60 V devices. Measurement results show that the proposed pulser can safely generate a bipolar pulse of -34.6 to 45 V, from a single 45 V supply voltage. The Tx/Rx switch blocks the HV bipolar pulse, resulting in less than 1.6 V at the input of the receiver. Acoustic measurements are performed connecting the pulser to CMUTs with 2 pF capacitance and 8 MHz center frequency. The variation of acoustic output pressures for different pulse shapes were simulated with the large signal CMUT model and compared with the experimental results for transmit pressure optimization. A potential implementation of the methods using MEMS fabrication methods is also described.

Acousto-Optic Catheter Tracking Sensor for Interventional MRI Procedures.

OBJECTIVE: The objective of this paper is to introduce an acousto-optic optical fiber sensor for tracking catheter position during interventional magnetic resonance imaging (MRI) to overcome RF induced heating of active markers. METHODS: The sensor uses a miniature coil coupled to a piezoelectric transducer, which is in turn mechanically connected to an optical fiber. The piezoelectric transducer converts the RF signal to acoustic waves in the optical fiber over a region including a fiber Bragg grating (FBG). The elastic waves in the fiber modulates the FBG geometry and hence the reflected light in the optical fiber. Since the coil is much smaller than the RF wavelength and the signal is transmitted on the dielectric optical fiber, the sensor effectively reduces RF induced heating risk. Proof of concept prototypes of the sensor are implemented using commercially available piezoelectric transducers and optical fibers with FBGs. The prototypes are characterized in a 1.5 T MRI system in comparison with an active tracking marker. RESULTS: Acousto-optical sensor shows linear response with flip angle and it can be used to detect signals from multiple coils for potential orientation detection. It has been successfully used to detect the position of a tacking coil in phantom in an imaging experiment. CONCLUSION: Acousto-optical sensing is demonstrated for tracking catheters during interventional MRI. Real-time operation of the sensor requires sensitivity improvements like using a narrow band FBG. SIGNIFICANCE: Acousto-optics provides a compact solution to sense RF signals in MRI with dielectric transmission lines.

Analysis and Design of High-Frequency 1-D CMUT Imaging Arrays in Noncollapsed Mode.

High-frequency ultrasound imaging arrays are important for a broad range of applications, from small animal imaging to photoacoustics. Capacitive micromachined ultrasonic transducer (CMUT) arrays are particularly attractive for these applications as low noise receiver electronics can be integrated for an overall improved performance. In this paper, we present a comprehensive analysis of high-frequency CMUT arrays based on an experimentally verified CMUT array simulation tool. The results obtained on an example, a 40-MHz 1-D CMUT array for intravascular imaging, are used to obtain key design insights and tradeoffs for receive only and pulse-echo imaging. For the receiver side, thermal mechanical current noise, plane wave pressure sensitivity, and pressure noise spectrum are extracted from simulations. Using these parameters, we find that the receiver performance of CMUT arrays can be close to an ideal piston, independent of gap thickness, and applied dc bias, when coupled to low noise electronics with arrays utilizing smaller membranes performing better. For pulse-echo imaging, thermal mechanical current noise limited signal-to-noise ratio is observed to be dependent on the maximum available voltage and gap thickness. In terms of bandwidth, we find that the Bragg resonance of the array, related to the fill factor, is a significant determinant of the high frequency limit and the fluid loaded single membrane resonance determines the lower limit. Based on these results, we present design guidelines requiring only fluid loaded single membrane simulations and membrane pitch to achieve a desired pulse-echo response. We also provide a design example and discuss limitations of the approach.

A Reduced-Wire ICE Catheter ASIC With Tx Beamforming and Rx Time-Division Multiplexing.

This paper presents a single chip reduced-wire active catheter application-specific integrated circuit (ASIC), equipped with programmable transmit (Tx) beamforming and receive (Rx) time-division multiplexing (TDM). The proposed front-end ASIC is designed for driving a 64-channel one-dimensional transducer array in intracardiac echocardiography (ICE) ultrasound catheters. The ASIC is implemented in 60 V 0.18-µm HV-BCD technology, integrating Tx beamformers with high voltage pulsers and Rx front end in the same chip, which occupies 2.6 × 11 mm2 that can fit in the catheter size of 9 F (<3 mm). The proposed system reduces the number of wires from >64 to only 22 by integrating Tx beamformer that is programmable using a single low-voltage differential signaling data line. In Rx mode, the system uses 8:1 TDM with direct digital demultiplexing providing raw channel data that enables dynamic Rx beamforming using individual array elements. This system has been successfully used for B-mode imaging on standard ultrasound phantom with 401 mW of average power consumption. The ASIC has a compact element pitch-matched layout, which is also compatible with capacitive micromachined ultrasound transducer on CMOS application. This system addresses cable number and dimensional restrictions in catheters to enable ICE imaging under magnetic resonance imaging by reducing radio frequency induced heating.

Analysis and Design of Capacitive Parametric Ultrasonic Transducers for Efficient Ultrasonic Power Transfer Based on a 1-D Lumped Model.

There is an increasing interest in wireless power transfer for medical implants, sensor networks, and consumer electronics. A passive capacitive parametric ultrasonic transducer (CPUT) can be suitable for these applications as it does not require a dc bias or a permanent charge. In this paper, we present a 1-D lumped parameter model of the CPUT to study its operation and investigate relevant design parameters for power transfer applications. The CPUT is modeled as an ultrasound-driven piston coupled to an RLC resonator resulting in a system of two coupled nonlinear ordinary differential equations. Simulink is used along with an analytical approximation of the system to obtain the voltage across the capacitor and displacement of the piston. Parametric resonance threshold and ultrasound-to-electrical conversion efficiency are evaluated, and the dependence of these performance metrics on the load resistance, input ultrasound intensity, forcing frequency, electrode coverage area, gap height, and the mechanical Q-factor are studied. Based on this analysis, design guidelines are proposed for highly efficient power transfer. Guided by these results, practical device designs are obtained through COMSOL simulations. Finally, the feasibility of using the CPUT in air is predicted to set the foundation for further research in ultrasonic wireless power transfer, energy harvesting, and sensing.

Supply-Doubled Pulse-Shaping High Voltage Pulser for CMUT Arrays.

A supply-doubled pulse-shaping high voltage (HV) pulser is presented for medical ultrasound imaging applications, particularly those that use capacitive micromachined ultrasonic transducers (CMUT). The pulser employs a bootstrap circuit combined with dynamically-biased stacked transistors, which allow HV operation above process limit without lowering device reliability. The new pulser overcomes supply voltage limitation of conventional unipolar pulsers by generating output signals that are almost twice the supply level. It also can generate three-level pulses to further optimize the transmit pressure signals. A proof-of-concept prototype has been implemented in 0.18-µm HV CMOS/DMOS technology with 60 V devices. Measurement results show that the HV pulser can safely generate controllable three-level pulses with up to 85 Vpp from 45 V supply. Acoustic measurements are conducted connecting the pulser to a CMUT with 2 pF capacitance and 8.3 MHz center frequency. The pulse shape has been adjusted for the CMUT under test to generate maximum pressure output and the results are in good agreement with a large signal CMUT model.

Low Temperature CMUT Fabrication Process with Dielectric Lift-off Membrane Support for Improved Reliability.

This paper reports an improved CMOS compatible low temperature sacrificial layer fabrication process for Capacitive Micromachined Ultrasonic Transducers (CMUTs). The process adds the fabrication step of silicon oxide evaporation which is followed by a lift-off step to define the membrane support area without a need for an extra mask. This simple addition improves reliability by reducing the electric field between the top and bottom electrodes everywhere except the moving membrane without affecting the vacuum gap thickness. Furthermore, the parasitic capacitance which degrades the CMUT receive performance is reduced. A 1-D CMUT array suitable for Intracardiac Echocardiography (ICE) imaging with 9MHz center frequency is fabricated using this method. Detailed electrical and acoustic testing indicates adequate performance of the devices for ICE in agreement with simulations. Long term output pressure testing with more than 2×1011 pulsing cycles and environmental testing demonstrate the efficacy of the approach for improved reliability as compared to devices without the additional membrane support layer.

Enhanced intracellular delivery via coordinated acoustically driven shear mechanoporation and electrophoretic insertion.

Delivery of large and structurally complex target molecules into cells is vital to the emerging areas of cellular modification and molecular therapy. Inadequacy of prevailing in vivo (viral) and in vitro (liposomal) gene transfer methods for delivery of proteins and a growing diversity of synthetic nanomaterials has encouraged development of alternative physical approaches. Efficacy of injury/diffusion-based delivery via shear mechanoporation is largely insensitive to cell type and target molecule; however, enhanced flexibility is typically accompanied by reduced gene transfer effectiveness. We detail a method to improve transfection efficiency through coordinated mechanical disruption of the cell membrane and electrophoretic insertion of DNA to the cell interior. An array of micromachined nozzles focuses ultrasonic pressure waves, creating a high-shear environment that promotes transient pore formation in membranes of transmitted cells. Acoustic Shear Poration (ASP) allows passive cytoplasmic delivery of small to large nongene macromolecules into established and primary cells at greater than 75% efficiency. Addition of an electrophoretic action enables active transport of target DNA molecules to substantially augment transfection efficiency of passive mechanoporation/diffusive delivery without affecting viability. This two-stage poration/insertion method preserves the compelling flexibility of shear-based delivery, yet substantially enhances capabilities for active transport and transfection of plasmid DNA.