Functional photoacoustic imaging: from nano- and micro- to macro-scale.

Functional photoacoustic imaging is a promising biological imaging technique that offers such unique benefits as scalable resolution and imaging depth, as well as the ability to provide functional information. At nanoscale, photoacoustic imaging has provided super-resolution images of the surface light absorption characteristics of materials and of single organelles in cells. At the microscopic and macroscopic scales. photoacoustic imaging techniques have precisely measured and quantified various physiological parameters, such as oxygen saturation, vessel morphology, blood flow, and the metabolic rate of oxygen, in both human and animal subjects. This comprehensive review provides an overview of functional photoacoustic imaging across multiple scales, from nano to macro, and highlights recent advances in technology developments and applications. Finally, the review surveys the future prospects of functional photoacoustic imaging in the biomedical field.

Two-step proximal gradient descent algorithm for photoacoustic signal unmixing.

Photoacoustic microscopy uses multiple wavelengths to measure concentrations of different absorbers. The speed of sound limits the shortest wavelength switching time to sub-microseconds, which is a bottleneck for high-speed broad-spectrum imaging. Via computational separation of overlapped signals, we can break the sound-speed limit on the wavelength switching time. This paper presents a new signal unmixing algorithm named two-step proximal gradient descent. It is advantageous in separating multiple wavelengths with long overlapping and high noise. In the simulation, we can unmix up to nine overlapped signals and successfully separate three overlapped signals with 12-ns delay and 15.9-dB signal-to-noise ratio. We apply this technique to separate three-wavelength photoacoustic images in microvessels. In vivo results show that the algorithm can successfully unmix overlapped multi-wavelength photoacoustic signals, and the unmixed data can improve accuracy in oxygen saturation imaging.

Optical-resolution photoacoustic microscopy with ultrafast dual-wavelength excitation.

Fast functional and molecular photoacoustic microscopy requires pulsed laser excitations at multiple wavelengths with enough pulse energy and short wavelength-switching time. Recent development of stimulated Raman scattering in optical fiber offers a low-cost laser source for multiwavelength photoacoustic imaging. In this approach, long fibers temporally separate different wavelengths via optical delay. The time delay between adjacent wavelengths may eventually limits the highest A-line rate. In addition, a long-time delay in fiber may limit the highest pulse energy, leading to poor image quality. In order to achieve high pulse energy and ultrafast dual-wavelength excitation, we present optical-resolution photoacoustic microscopy with ultrafast dual-wavelength excitation and a signal separation method. The signal separation method is validated in numerical simulation and phantom experiments. We show that when two photoacoustic signals are partially overlapped with a 50-ns delay, they can be recovered with 98% accuracy. We apply this ultrafast dual-wavelength excitation technique to in vivo OR-PAM. Results demonstrate that A-lines at two wavelengths can be successfully separated, and sO2 values can be reliably computed from the separated data. The ultrafast dual-wavelength excitation enables fast functional photoacoustic microscopy with negligible misalignment among different wavelengths and high pulse energy, which is important for in vivo imaging of microvascular dynamics.