The idea of a method of cost-effective upgrades from an acoustic resolution photoacoustic microscope to a triple-modality imaging system is validated using phantoms. The newly developed experimental setup is based on a diode pumped solid state laser coupled to a fiber bundle with a spherically focused polyvinylidene fluoride detector integrated into the center of a ring shaped optical illuminator. Each laser pulse illuminating the sample performs two functions. While the photons absorbed by the sample provide a measurable optoacoustic (OA) signal, the photons absorbed by the detector provide the measurable diffuse reflectometry (DR) signal from the sample and the probing ultrasonic (US) pulse. At a 3 mm imaging depth, the axial resolution of the OA/US modalities is 38 μm/26 μm, while the lateral resolution of the DR/OA/US modalities is 3.5 mm/50 μm/35 μm. The maximum acquisition rate of the trimodal DR/OA/US A-scans is 2 kHz.

An experimental setup for combined photoacoustic (PA) and optically mediated ultrasound (US) microscopy is presented. A spherically focused 35 MHz polyvinylidene fluoride (PVDF) ultrasonic detector with a numerical aperture of 0.28, a focal distance of 9 mm, and a bandwidth ($-6\mathrm{dB}$ level) of 24 MHz was used to obtain PA and US data with a 3 mm imaging depth. A fiber-optic system was employed to deliver laser excitation pulses from a tunable laser to the studied medium. A single optical pulse was used to form both PA and US A-scans. The probing US pulses were generated thermoelastically due to absorption of backscattered laser radiation by the metalized surface of a PVDF film.

We propose the use of thermoelastic (TE) excitation of an ultrasonic (US) detector by backscattered laser radiation as a means of upgrading a single-modality photoacoustic (PA) microscope to dual-modality PA/US imaging at minimal cost. The upgraded scanning head of our dual-modality microscope consists of a fiber bundle with 14 output arms and a 32MHz polyvinylidene difluoride (PVDF) detector with a 34 MHz bandwidth (−6 dB level), 12.7 mm focal length, and a 0.25 numerical aperture. A single optical pulse delivered through the fiber bundle to the biotissue being investigated, in combination with a metalized surface on the PVDF detector allows us to obtain both PA and US A-scans. To demonstrate the in vivo capabilities of the proposed method we present the results of bimodal imaging of the brain of a newborn rat, a mouse tail and a mouse tumor.

Cerebrovascular imaging of rodents is one of the trending applications of optoacoustics aimed at studying brain activity and pathology. Imaging of deep brain structures is often hindered by sub-optimal arrangement of the light delivery and acoustic detection systems. In our work we revisit the physics behind opto-acoustic signal generation for theoretical evaluation of optimal laser wavelengths to perform cerebrovascular optoacoustic angiography of rodents beyond the penetration barriers imposed by light diffusion in highly scattering and absorbing brain tissues. A comprehensive model based on diffusion approximation was developed to simulate optoacoustic signal generation using optical and acoustic parameters closely mimicking a typical murine brain. The model revealed three characteristic wavelength ranges in the visible and near-infrared spectra optimally suited for imaging cerebral vasculature of different size and depth. The theoretical conclusions are confirmed by numerical simulations while in vivo imaging experiments further validated the ability to accurately resolve brain vasculature at depths ranging between 0.7 and 7 mm.

Owing to its unique ability to capture volumetric tomographic information with a single light flash, optoacoustic (OA) tomography has recently demonstrated ultrafast imaging speeds ultimately limited by the ultrasound time-of-flight. The method’s scalability and the achievable spatial resolution are yet limited by the narrow bandwidth of piezo-composite arrays currently employed for OA signal detection. Here we report on the first implementation of high-density spherical array technology based on flexible polyvinylidene difluoride films featuring ultrawideband (0.3–40 MHz) sub mm² area elements, thus enabling real-time multi-scale volumetric imaging with 22–35 µm spatial resolution, superior image fidelity and over an order of magnitude signal-to-noise enhancement compared to piezo-composite equivalents. We further demonstrate five-dimensional (spectroscopic, time-resolved, volumetric) imaging capabilities by visualizing fast stimulus-evoked cerebral oxygenation changes in mice and performing real-time functional angiography of deep human micro-vasculature. The new technology thus leverages the true potential of OA for quantitative high-resolution visualization of rapid bio-dynamics across scales.

The letter discusses the opportunity for cost-effective use of conventional optoacoustic hardware to realize additional imaging modalities such as ultrasonic microscopy and diffuse optical reflectometry within the same laser pulse. Optoacoustic methods for deep biomedical visualization are based on pulsed laser illumination of the internal tissue layers with scattered photons, however some of the back-scattered photons can be absorbed by the optoacoustic detector. Thermoelastic extension of the detector’s surface provides a probing pulse for an ultrasonic modality while the measurement of the amplitude of the probing ultrasonic pulse allows estimation of the diffuse reflectance from the object under investigation.