Optical-resolution photoacoustic microscopy (OR-PAM) can provide functional, anatomical, and molecular images at micrometer level resolution with an imaging depth of less than 1 mm in tissue. However, the imaging speed of traditional OR-PAM is often low due to the point-by-point mechanical scanning and cannot capture time-sensitive dynamic information. In this work, we demonstrate a recent effort in improving the imaging speed of OR-PAM, using a newly developed water-immersible two-axis scanner. Driven by water-compatible electromagnetic actuation force, the new scanning mirror employs a novel torsion-bending mechanism to achieve fast 2D scanning. The torsion scanning along the fast-axis works in the resonant model, and the bending scanning along the slow-axis operate at the quasi-static mode. The scanning speed and scanning range along the two axes can be independently adjusted. Steered by the two-axis torsion-bending scanning mirror immersed in water, the focused excitation light and the generated acoustic wave can be confocally aligned over the entire imaging area. Thus, a high imaging speed can be achieved without sacrificing the detection sensitivity. Equipped with the torsion-bending scanner, the high-speed OR-PAM system has achieved a cross-sectional frame rate of 400 Hz, and a volumetric imaging speed of 1 Hz over a field of view of 1.5 × 2.5 mm². We have also demonstrated high-speed OR-PAM of the hemodynamic changes in response to pharmaceutical and physiological challenges in small animal models in vivo. We expect the torsion-bending scanner based OR-PAM will find matched biomedical studies of tissue dynamics.

We applied high‐speed photoacoustic microscopy (PAM) for both cortical microenvironment studies and dynamic brain studies, with micrometer‐level optical resolution and a millisecond‐level cross‐sectional imaging speed over a millimeter‐level field of view. We monitored blood flow redistribution in mini‐stroke mouse models and cerebral autoregulation induced by a vasoactive agent. Our results collectively suggest that high‐speed PAM is a promising tool for understanding dynamic neurophysiological phenomena, complementing conventional imaging modalities. magnified image

The conventional photoacoustic microscopy (PAM) system allows trade-offs between lateral resolution and imaging depth, limiting its applications in biological imaging in vivo. Here we present an integrated optical-resolution (OR) and acoustic-resolution (AR) multiscale PAM based on free-space light transmission and fast microelectromechanical systems (MEMS) scanning. The lateral resolution for OR is 4.9 µm, and the lateral resolution for AR is 114.5 µm. The maximum imaging depth for OR is 0.7 mm, and the maximum imaging depth for AR is 4.1 mm. The imaging speed can reach 50 k Alines per second. The high signal-to-noise ratios and wavelength throughput are achieved by delivering light via free-space, and the high speed is achieved by a MEMS scanning mirror. The blood vasculature from superficial skin to the deep tissue of a mouse leg was imaged in vivo using two different resolutions to demonstrate the multiscale imaging capability.

This Letter reports acoustic-resolution-photoacoustic microscopy (AR-PAM) based on a new optically transparent focused polyvinylidene fluoride (PVDF) transducer with a high acoustic numerical aperture (NA) of 0.64. Owing to the improved fabrication process, the new transducer has a much higher NA (0.64) than the previously reported low-NA transducer ( N A = 0.23 ). The acoustic center frequency and (pulse-echo) bandwidth are also increased to 36 and 44 MHz, respectively, which provides a 38 µm acoustic focal spot size and 210 µm acoustic depth of focus. For demonstration, AR-PAM was conducted on a twisted wire target in water and chicken breast tissue, and in vivo on a mouse tail. The imaging results show that high acoustic resolution and sensitivity can be achieved with a simple and compact setup to resolve the target at different depths. Such capabilities can be useful for the development of new AR-PAM systems for handheld, wearable, and even endoscopic imaging applications.