Visualizing tumor angiogenesis and boundary with polygon-scanning multiscale photoacoustic microscopy.

Recently, we developed an integrated optical-resolution (OR) and acoustic-resolution (AR) PAM, which has multiscale imaging capability using different resolutions. However, limited by the scanning method, a tradeoff exists between the imaging speed and field of view, which impedes its wider applications. Here, we present an improved multiscale PAM which achieves high-speed wide-field imaging based on a homemade polygon scanner. Encoder trigger mode was proposed to avoid jittering of the polygon scanner during imaging. Distortions caused by polygon scanning were analyzed theoretically and compared with traditional types of distortions in optical-scanning PAM. Then a depth correction method was proposed and verified to compensate for the distortions. System characterization of OR-PAM and AR-PAM was performed prior to in vivo imaging. Blood reperfusion of an in vivo mouse ear was imaged continuously to demonstrate the feasibility of the multiscale PAM for high-speed imaging. Results showed that the maximum B-scan rate could be 14.65 Hz in a fixed range of 10 mm. Compared with our previous multiscale system, the imaging speed of the improved system was increased by a factor of 12.35. In vivo imaging of a subcutaneously inoculated B-16 melanoma of a mouse was performed. Results showed that the blood vasculature around the melanoma could be resolved and the melanoma could be visualized at a depth up to 1.6 mm using the multiscale PAM.

Validation of photoacoustic/ultrasound dual imaging in evaluating blood oxygen saturation.

Photoacoustic imaging (PAI) was performed to evaluate oxygen saturation (sO2) of blood-mimicking phantoms, femoral arteries in beagles, and radial arteries in humans at various sO2 plateaus. The accuracy (root mean square error, RMSE) of PAI sO2 compared with reference sO2 was calculated. In blood-mimicking phantoms, PAI achieved an accuracy of 1.49% and a mean absolute error (MAE) of 1.09% within 25 mm depth, and good linearity (R = 0.968; p < 0.001) was obtained between PAI sO2 and reference sO2. In canine femoral arteries, PAI achieved an accuracy of 2.16% and an MAE of 1.58% within 8 mm depth (R = 0.965; p < 0.001). In human radial arteries, PAI achieved an accuracy of 3.97% and an MAE of 3.28% in depth from 4 to 14 mm (R = 0.892; p < 0.001). For PAI sO2 evaluation at different depths in healthy volunteers, the RMSE accuracy of PAI sO2 increased from 2.66% to 24.96% with depth increasing from 4 to 14 mm. Through the multiscale method, we confirmed the feasibility of the hand-held photoacoustic/ultrasound (PA/US) in evaluating sO2. These results demonstrate the potential clinical value of PAI in evaluating blood sO2. Consequently, protocols for verifying the feasibility of medical devices based on PAI may be established.

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 10.365 µm.

[Photoacoustic imaging study on spatial structure of microvessels around acupoints in mice with acute myocardial ischemia].

OBJECTIVE: To observe the changes of microvascular structure of acupoints caused by myocardial ischemia, so as to explore the application of photoacoustic imaging technology in the research of acupoint sensitization. METHODS: Twelve BALB/c mice were randomly divided into normal, sham operation and acute myocardial ischemia (AMI) model groups, with 4 mice in each group. AMI model was established by ligating the left anterior descending coronary artery. Electrocardiogram (ECG) was recorded by physiological signal acquisition system at 12 h and on the 14th day after modeling, and serum cardiac troponin T (cTnT) and creatine kinase isoenzyme MB (CK-MB) levels were detected by enzyme-linked immunosorbent assay. The microvascular structure changes of acupoints "Feishu"(BL13), "Jueyinshu"(BL14), "Quze"(PC3) and "Chize"(LU5) were observed by photoacoustic imaging technology, and distance (DM), inflection count metric (ICM), sum of angle metric(SOAM)and microvessel density (MVD) were calculated by microvascular quantification algorithm. RESULTS: Compared with the normal and sham operation groups, the ST segment of ECG was obviously elevated, serum cTnT and CK-MB were significantly increased in AMI model group at 12 h and on the 14th day after AMI (P<0.01). The ICM of BL14 in AMI model group was significantly decreased on the 14th day than that on the 7th day after AMI. Compared with the normal group, the ICM of BL14 was significantly increased in AMI model group on the 7th day after AMI(P<0.05). There were no significant changes in DM, ICM, SOAM and MVD at other acupoints on the 7th and 14th day (P>0.05) among the three groups. CONCLUSION: The change of ICM may be one of the characteristics of acupoint sensitization and photoacoustic imaging technology can be used to study the structure of acupoint microvessels.

Photoacoustic visualization of the fluence rate dependence of photodynamic therapy.

This study investigates the fluence rate effect, an essential modulating mechanism of photodynamic therapy (PDT), by using photoacoustic imaging method. To the best of our knowledge, this is the first time that the fluence rate dependence is investigated at a microscopic scale, as opposed to previous studies that are based on tumor growth/necrosis or animal surviving rate. This micro-scale examination enables subtle biological responses, including the vascular damage and the self-healing response, to be studied. Our results reveal the correlations between fluence rate and PDT efficacy/self-healing magnitude, indicating that vascular injuries induced by high fluence rates are more likely to recover and by low fluence rates (≤126 mW/cm2) are more likely to be permanent. There exists a turning point of fluence rate (314 mW/cm2), above which PDT practically produces no permanent therapeutic effect and damaged vessels can fully recover. These findings have practical significance in clinical setting. For cancer-related diseases, the 'effective fluence rate' is useful to provoke permanent destruction of tumor vasculature. Likewise, the 'non effective range' can be applied when PDT is used in applications such as opening the blood brain barrier to avoid permanent brain damage.