A promising approach for quantifying focal stroke modeling and assessing stroke progression:optical resolution photoacoustic microscopy photothrombosis

To investigate the mechanisms underlying the onset and progression of ischemic stroke,some methods have been proposed that can simultaneously monitor and create embolisms in the animal cerebral cortex.However,these methods often require complex systems and the effect of age on cerebral embolism has not been adequately studied,although ischemic stroke is strongly age-related.In this study,we propose an optical-resolution photoacoustic microscopy-based visualized photothrombosis methodology to create and monitor ischemic stroke in mice simultaneously using a 532 nm pulsed laser.We observed the molding process in mice of different ages and presented age-dependent vascular embolism differentiation.Moreover,we integrated optical coherence tomography angiography to investigate age-associated trends in cerebrovascular variability following a stroke.Our imaging data and quantitative analyses underscore the differential cerebrovascular responses to stroke in mice of different ages,thereby highlighting the technique's potential for evaluating cerebrovascular health and unraveling age-related mechanisms involved in ischemic strokes.

Photoacoustic microscopy based on transparent piezoelectric ultrasound transducers

Photoacoustic microscopy(PAM),due to its deep penetration depth and high contrast,is playing an increasingly important role in biomedical imaging.PAM imaging systems equipped with conventional ultrasound transducers have demonstrated excellent imaging performance.However,these opaque ultrasonic transducers bring some constraints to the further development and application of PAM,such as complex optical path,bulky size,and difficult to integrate with other modalities.To overcome these problems,ultrasonic transducers with high optical transparency have appeared.At present,transparent ultrasonic transducers are divided into optical-based and acoustic-based sensors.In this paper,we mainly describe the acoustic-based piezoelectric transparent transducers in detail,of which the research advances in PAM applications are reviewed.In addition,the potential challenges and developments of transparent transducers in PAM are also demonstrated.

All-optically integrated photoacoustic and optical coherence tomography:A review

All-optically integrated photoacoustic(PA)and optical coherence tomography(OCT)dualmode imaging technology that could o®er comprehensive pathological information for accurate diagnosis in clinic has gradually become a promising imaging technology in the aspect of biomedical imaging during the recent years.This review refers to the technology aspects of alloptical PA detection and system evolution of optically integrated PA and OCT,including Michelson interferometer dual-mode imaging system,Fabry–Perot(FP)interferometer dualmode imaging system and Mach–Zehnder interferometer dual-mode imaging system.It is believed that the optically integrated PA and OCT has great potential applications in biomedical imaging.

High-speed all-optic optical coherence tomography and photoacoustic microscopy dual-modal system for microcirculation evaluation

We propose a high-speed all-optic dual-modal system that integrates spectral domain optical coherence tomography and photoacoustic microscopy(PAM).A 3*3 coupler-based interfer-ometer is used to remotely detect the surface vibration caused by photoacoustic(PA)waves.Three outputs of the interferometer are acquired simultaneously with a multi-channel data ac-quisition card.One channel data with the highest PA signal detection sensitivity is selected for sensitivity compensation.Experiment on the phantom demonstrates that the proposed method can sucessfully compensate for the loss of intensity caused by sensitivity variation.The imaging speed of the PAM is improved compared to our previous system.The total time to image a sample with 256×256 pixels is~20s.Using the proposed system,the microvasculature in the mouse auricle is visualized and the blood flow state is accessed.

Non-invasive and low-artifact in vivo brain imaging by using a scanning acoustic-photoacoustic dual mode microscopy

Photoacoustic imaging is a potential candidate for in vivo brain imaging,whereas,its imaging performance could be degraded by inhomogeneous multi-layered media,consisted of scalp and skull.In this work,we propose a low-artifact photoacoustic microscopy(LAPAM)scheme,which combines conventional acoustic-resolution photoacoustic microscopy with scanning acoustic microscopy to suppress the reflection artifacts induced by multi-layers.Based on similar propagation characteristics of photoacoustic signals and ultrasonic echoes,the ultrasonic echoes can be employed as the filters to suppress the reflection artifacts to obtain low-artifact photoacoustic images.Phantom experiment is used to validate the effectiveness of this method.Furthermore,LAPAM is applied for in-vivo imaging mouse brain without removing the scalp and the skull.Experimental results show that the proposed method successfully achieves the low-artifact brain image,which demonstrates the practical applicability of LAPAM.This work might improve the photoacoustic imaging quality in many biomedical applications which involve tissues with complex acoustic properties,such as brain imaging through scalp and skull.

Concurrent photoacoustic and ultrasound microscopy with a coaxial dual-element ultrasonic transducer

Simultaneous photoacoustic and ultrasound(PAUS)imaging has attracted increasing attention in biomedical research to probe the optical and mechanical properties of tissue.However,the resolution for majority of the existing PAUS systems is on the order of 1 mm as the majority are designed for clinical use with low-frequency US detection.Here we developed a concurrent PAUS microscopy that consists of optical-resolution photoacoustic microscopy(OR-PAM)and high-frequency US pulse-echo imaging.This dual-modality system utilizes a novel coaxial dual-element ultrasonic transducer(DE-UST)and provides anatomical and functional information with complementary contrast mechanisms,achieving a spatial resolution of 7μm for PA imaging and 106μm for US imaging.We performed phantom studies to validate the system’s performance.The vasculature of a mouse’s hind paw was imaged to demonstrate the potential of this hybrid system for biomedical applications.

A SIMULTANEOUS MULTI-PROBE DETECTION LABEL-FREE OPTICAL-RESOLUTION PHOTOACOUSTIC MICROSCOPY TECHNIQUE BASED ON MICROCAVITY TRANSDUCER

We demonstrate the feasibility of simultancous multi-probe detection for an optcal-resolution photoacoustic microscopy(OR-PAM)system.OR-P AM has elicited the attention of biomedical imaging researchers because of its optical absorption contrast and high spatial resolution with great imaging depth.OR-PAM allows label-free and noninvasive imaging by maximizing the optical absorption of endogenous biomolecules.However,given the inadequate absoption of some biomolcules,detection sensitivity at the same incident intensity requires improvement.In this study,a modulated continuous wave with power density less than 3mW/cm^(2)(1/4 of the ANSI safety limit)excited the weak photoacoustic(PA)signals of biological cells.A microcavity traneducer is developed based on the bulk modulus of gas five orders of magnitude lower than that of solid;air pressure variation is inversely proportional to cavity volume at the same temperature increase.Considering that a PA wave expands in various directions,detecting PA signals from different positions and adding them together can increase detection sensitivity and signal-to-noise ratio.Therefore,we employ four detectors to acquire tiny PA signals simul-taneously.Experimental results show that the developed OR-PAM system allows the label-free imaging of cells with weak optical absorption.

In vivo evaluation of laser-induced choroidal neovascularization in rats simultaneously using optical coherence tomography and photoacoustic microscopy

Determination of the precise location and the degree of the Choroidal neovascularization(CNV)lesion is essential for diagnosation Neovascular age-related macular degeneration(AMD)and evaluation the efficacy of treatment.Noninvasive imaging techniques with specific contrast for CNV evaluation are demanded.In this paper,two noninvasive imaging techniques,namely Optical coherence tomography(OCT)and Photoacoustic microscopy(PAM),are combined to provide specific detection of CNV for their complimentary contrast mechanisms.In vivo time-serial evaluation of Laser-induced CNV in rats is present at days 1,3,5,7,14,21 after laser photocoagulation is applied to the rat fundus.Both OCT and PAM show that the CNV increases to its maximum at day 7 and decreases at day 14.Quantification of CNV area and CNV thickness is given.The dual-modal information of CNV is consistent with the histologic evaluation by hematoxylin and eosin(H&E)staining.

Spatial weight matrix in dimensionality reduction reconstruction for microelectromechanical system-based photoacoustic microscopy

A micro-electromechanical system(MEMS)scanning mirror accelerates the raster scanning of optical-resolution photoacoustic microscopy(OR-PAM).However,the nonlinear tilt angular-voltage characteristic of a MEMS mirror introduces distortion into the maximum back-projection image.Moreover,the size of the airy disk,ultrasonic sensor properties,and thermal effects decrease the resolution.Thus,in this study,we proposed a spatial weight matrix(SWM)with a dimensionality reduction for image reconstruction.The three-layer SWM contains the invariable information of the system,which includes a spatial dependent distortion correction and 3D deconvolution.We employed an ordinal-valued Markov random field and the Harris Stephen algorithm,as well as a modified delay-and-sum method during a time reversal.The results from the experiments and a quantitative analysis demonstrate that images can be effectively reconstructed using an SWM;this is also true for severely distorted images.The index of the mutual information between the reference images and registered images was 70.33 times higher than the initial index,on average.Moreover,the peak signal-to-noise ratio was increased by 17.08%after 3D deconvolution.This accomplishment offers a practical approach to image reconstruction and a promising method to achieve a real-time distortion correction for MEMS-based OR-PAM.

Deep-learning-based motion-correction algorithm in optical resolution photoacoustic microscopy

In this study,we propose a deep-learning-based method to correct motion artifacts in optical resolution photoacoustic microscopy(OR-PAM).The method is a convolutional neural network that establishes an end-to-end map from input raw data with motion artifacts to output corrected images.First,we performed simulation studies to evaluate the feasibility and effectiveness of the proposed method.Second,we employed this method to process images of rat brain vessels with multiple motion artifacts to evaluate its performance for in vivo applications.The results demonstrate that this method works well for both large blood vessels and capillary networks.In comparison with traditional methods,the proposed method in this study can be easily modified to satisfy different scenarios of motion corrections in OR-PAM by revising the training sets.

Reflection-mode photoacoustic microscopy using a hollow focused ultrasound transducer for in vivo imaging of blood vessels

A reflection-mode photoacoustic microscope using a hollow focused ultrasound transducer is developed for highresolution in vivo imaging.A confocal structure of the laser and the ultrasound is used to improve the system resolution.The axial and lateral resolutions of the system are measured to be ～ 32 μm and ～ 58 μm,respectively.Ex vivo and in vivo modes are tested to validate the imaging capability of the photoacoustic microscope.The adjacent vein and artery can be seen clearly from the reconstructed photoacoustic images.The results demonstrate that the reflectionmode photoacoustic microscope can be used for high-resolution imaging of micro-blood vessels,which would be of great benefit for monitoring the neovascularization in tumor angiogenesis.

Fiber laser technologies for photoacoustic microscopy

Fiber laser technology has experienced a rapid growth over the past decade owing to increased applications in precision measurement and optical testing,medical care,and industrial applications,including laser welding,cleaning,and manufacturing.A fiber laser can output laser pulses with high energy,a high repetition rate,a controllable wavelength,low noise,and good beam quality,making it applicable in photoacoustic imaging.Herein,recent developments in fiber-laser-based photoacoustic microscopy(PAM)are reviewed.Multispectral PAM can be used to image oxygen saturation or lipid-rich biological tissues by applying a Q-switched fiber laser,a stimulated Raman scattering-based laser source,or a fiber-based supercontinuum source for photoacoustic excitation.PAM can also incorporate a single-mode fiber laser cavity as a high-sensitivity ultrasound sensor by measuring the acoustically induced lasing-frequency shift.Because of their small size and high flexibility,compact head-mounted,wearable,or hand-held imaging modalities and better photoacoustic endoscopes can be enabled using fiber-laser-based PAM.

A microvascular image analysis method for optical-resolution photoacoustic microscopy

Optical-resolution photoacoustic microscopy(OR-PAM)has been shown to be an excellent tool for high-resolution imaging of microvasculature,and quantitative analysis of the microvascula-ture can provide valuable information for the early diagnosis and treatment of various vascular-related diseases.In order to address the characteristics of weak signals,discontinuity and small diameters in photoacoustic microvascular images,we propose a method adaptive to the micro-vascular segmentation in photoacoustic images,including Hessian matrix enhancement and the morphological connection operators.The accuracy of our vascular segmentation method is quantitatively evaluated by the multiple criteria.To obtain more precise and continuous mi-crovascular skeletons,an improved skeleton extraction framework based on the multistencil fast marching(MSFM)method is developed.We carried out in vivo OR-PAM microvascular imaging in mouse ears and subcutaneous hepatoma tumor model to verify the correctness and superiority of our proposed method.Compared with the previous methods,our proposed method can extract the microvascular network more completely,continuously and accurately,and provide an ef-fective solution for the quantitative analysis of photoacoustic microvascular images with many small branches.

High-speed high-resolution laser diode-based photoacoustic microscopy for in vivo microvasculature imaging

Laser diodes(LDs)have been considered as cost-effective and compact excitation sources to overcome the requirement of costly and bulky pulsed laser sources that are commonly used in photoacoustic microscopy(PAM).However,the spatial resolution and/or imaging speed of previously reported LD-based PAM systems have not been optimized simultaneously.In this paper,we developed a high-speed and high-resolution LD-based PAM system using a continuous wave LD,operating at a pulsed mode,with a repetition rate of 30 kHz,as an excitation source.A hybrid scanning mechanism that synchronizes a one-dimensional galvanometer mirror and a two-dimensional motorized stage is applied to achieve a fast imaging capability without signal averaging due to the high signal-to-noise ratio.By optimizing the optical system,a high lateral resolution of 4.8μm has been achieved.In vivo microvasculature imaging of a mouse ear has been demonstrated to show the high performance of our LD-based PAM system.

Simulation Platform of Multi-Pulse Photoacoustic Microscopy Based on K-Space Pseudospectral Method

Photoacoustic imaging has emerged in the past decades. Compared with the traditional medical imaging mode, it has better imaging performance and has great development potential in the field of biological imaging. In traditional photoacoustic microscopy, a single laser pulse is generally used to irradiate the sample to produce photoacoustic signal. And sig-nal-to-noise ratio (SNR) is a very important indicator for photoacoustic im-aging. In order to obtain the image with high SNR, multiple acquisition or increasing laser pulse energy is usually adopted. The former will lead to slower imaging speed, and the latter will lead to photobleaching or pho-totoxicity. Here, we propose multi-pulse photoacoustic microscopy, the photoacoustic signals were stimulated sequentially using multiple laser pulses in each A line data acquisition. In order to verify the feasibility of this method, a multi-pulse photoacoustic imaging simulation platform is established using k-Wave toolbox. The performance of multi-pulse photo-acoustic imaging is verified through the three scanning modes of photoa-coustic microscopy A-scan, B-scan, and C-scan. The results indicate that the SNR is proportion to the number of laser pulses used, high SNR can be achieved by low-energy laser pulse. This work will help to expand the ap-plication of photoacoustic imaging.

Flexible fiber-laser ultrasound sensor for multiscale photoacoustic imaging

Photoacoustic imag ing(PAI)is a nonin vasive biomedical imag ing tech no logy capable of multiscale imag ing of biological samples from orga ns dow n to cells.Multiscale PAI requires differe nt ultraso und tra nsducers that are flat or focused because the current widely-used piezoelectric transducers are rigid and lack the flexibility to tune their spatial ultrasound responses.Inspired by the rapidly-developing flexible photonics,we exploited the inherent flexibility and low-loss features of optical fibers to develop a flexible fiber-laser ultrasound sensor(FUS)for multiscale PAI.By simply bending the fiber laser from straight to curved geometry,the spatial ultraso und resp onse of the FUS can be tuned for both wide-view optical-resolution photoacoustic microscopy at optical diffraction-limited depth(~1 mm)and photoacoustic computed tomography at optical dissipation-limited depth of several centimeters.A radio-frequency demodulation was employed to get the readout of the beat frequency variation of two orthogonal polarization modes in the FUS output,which ensures low-noise and stable ultrasound detection.Compared to traditional piezoelectrical transducers with fixed ultrasound responses once manufactured,the flexible FUS provides the freedom to design multiscale PAI modalities including wearable microscope,intravascular endoscopy,and portable tomography system,which is attractive to fundamental biologic-al/medical studies and clinical applications.

Recent developments in photoacoustic imaging and sensing for nondestructive testing and evaluation

Photoacoustic(PA)imaging has been widely used in biomedical research and preclinical studies during the past two decades.It has also been explored for nondestructive testing and evaluation(NDT/E)and for industrial applications.This paper describes the basic principles of PA technology for NDT/E and its applications in recent years.PA technology for NDT/E includes the use of a modulated continuous-wave laser and a pulsed laser for PA wave excitation,PA-generated ultrasonic waves,and all-optical PA wave excitation and detection.PA technology for NDT/E has demonstrated broad applications,including the imaging of railway cracks and defects,the imaging of Li metal batteries,the measurements of the porosity and Young’s modulus,the detection of defects and damage in silicon wafers,and a visualization of underdrawings in paintings.

Low-cost photoacoustic imaging systems based on laser diode and light-emitting diode excitation

Photoacoustic imaging,an emerging biomedical imaging modality,holds great promise for preclinical and clinical researches.It combines the high optical contrast and high ultrasound resolution by converting laser excitation into ultrasonic emission.In order to generate photoacoustic signal e±-ciently,bulky Q-switched solid-state laser systems are most commonly used as excitation sources and hence limit its commercialization.As an alternative,the miniaturized semiconductor laser system has the advantages of being inexpensive,compact,and robust,which makes a signi¯cant e®ect on production-forming design.It is also desirable to obtain a wavelength in a wide range from visible to nearinfrared spectrum for multispectral applications.Focussing on practical aspect,this paper reviews the state-of-the-art developments of low-cost photoacoustic system with laser diode and light-emitting diode excitation source and highlights a few representative installations in the past decade.

Chronic cranial window for photoacoustic imaging: a mini review

Photoacoustic(PA)microscopy is being increasingly used to visualize the microcirculation of the brain cortex at the micron level in living rodents.By combining it with long-term cranial window techniques,vasculature can be monitored over a period of days extending to months through a field of view.To fulfill the requirements of long-term in vivo PA imaging,the cranial window must involve a simple and rapid surgical procedure,biological compatibility,and sufficient optical-acoustic transparency,which are major challenges.Recently,several cranial window techniques have been reported for longitudinal PA imaging.Here,the development of chronic cranial windows for PA imaging is reviewed and its technical details are discussed,including window installation,imaging quality,and longitudinal stability.

Recovery of photoacoustic images based on accurate ultrasound positioning

Photoacoustic microscopy is an in vivo imaging technology based on the photoacoustic effect.It is widely used in various biomedical studies because it can provide high-resolution images while being label-free,safe,and harmless to biological tissue.Polygon-scanning is an effective scanning method in photoacoustic microscopy that can realize fast imaging of biological tissue with a large field of view.However,in polygon-scanning,fluctuations of the rotating motor speed and the geometric error of the rotating mirror cause image distortions,which seriously affect the photoacoustic-microscopy imaging quality.To improve the image quality of photoacoustic microscopy using polygon-scanning,an image correction method is proposed based on accurate ultrasound positioning.In this method,the photoacoustic and ultrasound imaging data of the sample are simultaneously obtained,and the angle information of each mirror used in the polygon-scanning is extracted from the ultrasonic data to correct the photoacoustic images.Experimental results show that the proposed method can significantly reduce image distortions in photoacoustic microscopy,with the image dislocation offset decreasing from 24.774 to 10.365μm.