Noninvasive Quantification of Contractile Dynamics in Cardiac Cells, Spheroids, and Organs-on-a-Chip Using High-Frequency Ultrasound.

Cell-based models that mimic in vivo heart physiology are poised to make significant advances in cardiac disease modeling and drug discovery. In these systems, cardiomyocyte (CM) contractility is an important functional metric, but current measurement methods are inaccurate and low-throughput or require complex setups. To address this need, we developed a standalone noninvasive, label-free ultrasound technique operating at 40-200 MHz to measure the contractile kinetics of cardiac models, ranging from single adult CMs to 3D microtissue constructs in standard cell culture formats.

Air-Coupled Photoacoustic Detection of Airborne Particulates.

In this study, we present a novel method to detect airborne particulates using air-coupled photoacoustics, with a goal toward detecting viral content in respiratory droplets. The peak photoacoustic frequency emitted from micrometer-sized particulates is over 1000 MHz, but at this frequency, the signals are highly attenuated in air. Measurements were taken using a thin planar absorber and ultrasound transducers with peak sensitivity between 50 kHz and 2000 kHz and a 532 nm pulsed laser to determine the optimum detection frequency.

Assessing engineered tissues and biomaterials using ultrasound imaging: In vitro and in vivo applications.

Quantitative assessment of the structural, functional, and mechanical properties of engineered tissues and biomaterials is fundamental to their development for regenerative medicine applications. Ultrasound (US) imaging is a non-invasive, non-destructive, and cost-effective technique capable of longitudinal and quantitative monitoring of tissue structure and function across centimeter to sub-micron length scales. Here we present the fundamentals of US to contextualize its application for the assessment of biomaterials and engineered tissues, both in vivo and in vitro.

High-frequency quantitative ultrasound for the assessment of the acoustic properties of engineered tissues in vitro.

Acoustic properties of biomaterials and engineered tissues reflect their structure and cellularity. High-frequency ultrasound (US) can non-invasively characterize and monitor these properties with sub-millimetre resolution. We present an approach to estimate the speed of sound, acoustic impedance, and acoustic attenuation of cell-laden hydrogels that accounts for frequency-dependent effects of attenuation in coupling media, hydrogel thickness, and interfacial transmission/reflection coefficients of US waves, all of which can bias attenuation estimates.

Thrombin-responsive engineered nanoexcavator with full-thickness infiltration capability for pharmaceutical-free deep venous thrombosis theranostics.

Although nanotechnology has shown great promise for treating multiple vascular diseases in recent years, simultaneous noninvasive detection and efficient dissolution of deep venous thrombosis (DVT) still remains challenging. In particular, long blockage areas and large thrombus thicknesses in DVT cause enormous difficulties for site-specific deep-seated thrombus theranostics. Therefore, based on the unique components of DVT, the novel concept of a thrombin-responsive full-thickness infiltration nonpharmaceutical nanoplatform for DVT theranostics is proposed here.

Insights into photoacoustic speckle and applications in tumor characterization.

In ultrasound imaging, fully-developed speckle arises from the spatiotemporal superposition of pressure waves backscattered by randomly distributed scatterers. Speckle appearance is affected by the imaging system characteristics (lateral and axial resolution) and the random-like nature of the underlying tissue structure. In this work, we examine speckle formation in acoustic-resolution photoacoustic (PA) imaging using simulations and experiments.

Optical and photoacoustic radiofrequency spectroscopic analysis for detecting red blood cell death.

Under stress, red blood cells (RBCs) undergo programmed cell death (eryptosis). One of the signaling molecules for eryptosis, sphingomyelinase (SMase), plays an important role in monitoring the efficacy of vascular targeted cancer therapy. The high optical absorption of erythrocytes coupled with the changes of eryptotic RBCs makes RBCs ideal targets for the photoacoustic (PA) detection and characterization of vascular treatments.

Sizing biological cells using a microfluidic acoustic flow cytometer.

We describe a new technique that combines ultrasound and microfluidics to rapidly size and count cells in a high-throughput and label-free fashion. Using 3D hydrodynamic flow focusing, cells are streamed single file through an ultrasound beam where ultrasound scattering events from each individual cell are acquired. The ultrasound operates at a center frequency of 375 MHz with a wavelength of 4 μm; when the ultrasound wavelength is similar to the size of a scatterer, the power spectra of the backscattered ultrasound waves have distinct features at specific frequencies that are directly related to the cell size.

Simultaneous acoustic and photoacoustic microfluidic flow cytometry for label-free analysis.

We developed a label-free microfluidic acoustic flow cytometer (AFC) based on interleaved detection of ultrasound backscatter and photoacoustic waves from individual cells and particles flowing through a microfluidic channel. The AFC uses ultra-high frequency ultrasound, which has a center frequency of 375 MHz, corresponding to a wavelength of 4 μm, and a nanosecondpulsed laser, to detect individual cells. We validate the AFC by using it to count different color polystyrene microparticles and comparing the results to data from fluorescence-activated cell sorting (FACS).

Triplex micron-resolution acoustic, photoacoustic, and optical transmission microscopy via photoacoustic radiometry.

We present a new sensing technique, termed photoacoustic radiometry (PAR), for mapping the optical attenuation properties of a sample. In PAR, laser pulses attenuated via transmission through the sample impinge on the ultrasound transducer and generate a photoacoustic (PA) signal within it. Spatial variation of the optical attenuation properties of the sample influences the amplitude of the PAR signal, providing image contrast.

A Laser-Activated Biocompatible Theranostic Nanoagent for Targeted Multimodal Imaging and Photothermal Therapy.

Unlabelled: Multifunctional nanoparticles have been reported for cancer detection and treatment currently. However, the accurate diagnosis and efficient treatment for tumors are still not satisfied. Here we report on the development of targeted phase change multimodal polymeric nanoparticles for the imaging and treatment of HER2-positive breast cancer.

High frequency ultrasound imaging and simulations of sea urchin oocytes.

High frequency ultrasound backscatter signals from sea urchin oocytes were measured using a 40 MHz transducer and compared to numerical simulations. The Faran scattering model was used to calculate the ultrasound scattered from single oocytes in suspension. The urchin oocytes are non-nucleated with uniform size and biomechanical properties; the backscatter from each cell is similar and easy to simulate, unlike typical nucleated mammalian cells.

Low-power noncontact photoacoustic microscope for bioimaging applications.

An inexpensive noncontact photoacoustic (PA) imaging system using a low-power continuous wave laser and a kilohertz-range microphone has been developed. The system operates in both optical and PA imaging modes and is designed to be compatible with conventional optical microscopes. Aqueous coupling fluids are not required for the detection of the PA signals; air is used as the coupling medium.

Biodegradable polymeric nanoparticles containing gold nanoparticles and Paclitaxel for cancer imaging and drug delivery using photoacoustic methods.

In this study, optical-triggered multifunctional theranostic agents for photoacoustic/fluorescent imaging and cancer therapy have been developed. This system consists of a perfluorohexane liquid and gold nanoparticles (GNPs) in the core, stabilized by a Poly (lactide-co-glycolic acid) (PLGA) polymer shell. When cancer cells containing PLGA-GNPs were exposed to laser pulses, cell viability decreased due to the vaporization of the particles in and around the cells.

High resolution ultrasound and photoacoustic imaging of single cells.

High resolution ultrasound and photoacoustic images of stained neutrophils, lymphocytes and monocytes from a blood smear were acquired using a combined acoustic/photoacoustic microscope. Photoacoustic images were created using a pulsed 532 nm laser that was coupled to a single mode fiber to produce output wavelengths from 532 nm to 620 nm via stimulated Raman scattering. The excitation wavelength was selected using optical filters and focused onto the sample using a 20× objective.

Properties of cells through life and death - an acoustic microscopy investigation.

Current methods to evaluate the status of a cell are largely focused on fluorescent identification of molecular biomarkers. The invasive nature of these methods - requiring either fixation, chemical dyes, genetic alteration, or a combination of these - prevents subsequent analysis of samples. In light of this limitation, studies have considered the use of physical markers to differentiate cell stages.

High-Frequency Acoustic Impedance Imaging of Cancer Cells.

Variations in the acoustic impedance throughout cells and tissue can be used to gain insight into cellular microstructures and the physiologic state of the cell. Ultrasound imaging can be used to create a map of the acoustic impedance, on which fluctuations can be used to help identify the dominant ultrasound scattering source in cells, providing information for ultrasound tissue characterization. The physiologic state of a cell can be inferred from the average acoustic impedance values, as many cellular physiologic changes are linked to an alteration in their mechanical properties.

Classification of blood cells and tumor cells using label-free ultrasound and photoacoustics.

A label-free method that can identify cells in a blood sample using high frequency photoacoustic and ultrasound signals is demonstrated. When the wavelength of the ultrasound or photoacoustic wave is similar to the size of a single cell (frequencies of 100-500 MHz), unique periodic features occur within the ultrasound and photoacoustic power spectrum that depend on the cell size, structure, and morphology. These spectral features can be used to identify different cell types present in blood, such as red blood cells (RBCs), white blood cells (WBCs), and circulating tumor cells.

High frequency label-free photoacoustic microscopy of single cells.

Photoacoustic measurements of melanoma cells and red blood cells (RBCs) using ultra-high frequency (UHF) wide-bandwidth transducers are reported. In this detection system, the resolution typically depends on the parameters of the receiving transducer, and not the focus of the laser. A single melanoma cell was imaged with 200, 375 and 1200 MHz transducers.

Modeling photoacoustic spectral features of micron-sized particles.

The photoacoustic signal generated from particles when irradiated by light is determined by attributes of the particle such as the size, speed of sound, morphology and the optical absorption coefficient. Unique features such as periodically varying minima and maxima are observed throughout the photoacoustic signal power spectrum, where the periodicity depends on these physical attributes. The frequency content of the photoacoustic signals can be used to obtain the physical attributes of unknown particles by comparison to analytical solutions of homogeneous symmetric geometric structures, such as spheres.

Probing red blood cell morphology using high-frequency photoacoustics.

A method that can rapidly quantify variations in the morphology of single red blood cells (RBCs) using light and sound is presented. When irradiated with a laser pulse, an RBC absorbs the optical energy and emits an ultrasonic pressure wave called a photoacoustic wave. The power spectrum of the resulting photoacoustic wave contains distinctive features that can be used to identify the RBC size and morphology.

Acoustic and photoacoustic characterization of micron-sized perfluorocarbon emulsions.

Perfluorocarbon droplets containing nanoparticles (NPs) have recently been investigated as theranostic and dual-mode contrast agents. These droplets can be vaporized via laser irradiation or used as photoacoustic contrast agents below the vaporization threshold. This study investigates the photoacoustic mechanism of NP-loaded droplets using photoacoustic frequencies between 100 and 1000 MHz, where distinct spectral features are observed that are related to the droplet composition.

Measuring the mechanical properties of cells using acoustic microscopy.

Time resolved acoustic microscopy was used to investigate the mechanical properties of MCF7 breast cancer cells at 375 MHz. The thickness, speed of sound, acoustic impedance, density, bulk modulus and attenuation were calculated using the amplitude and time of the ultrasound backscatter from the cellular membrane and substrate. The technique was first validated by measuring the mechanical properties of a material with well characterized properties, polyvinylidene fluoride (PVDF).