Microvascular quantification based on contour-scanning photoacoustic microscopy

Accurate quantification of microvasculature remains of interest in fundamental pathophysiological studies and clinical trials. Current photoacoustic microscopy can noninvasively quantify properties of the microvasculature, including vessel density and diameter, with a high spatial resolution. However, the depth range of focus (i.e., focal zone) of optical-resolution photoacoustic microscopy (OR-PAM) is often insufficient to encompass the depth variations of features of interest—such as blood vessels—due to uneven tissue surfaces. Thus, time-consuming image acquisitions at multiple different focal planes are required to maintain the region of interest in the focal zone. We have developed continuous three-dimensional motorized contour-scanning OR-PAM, which enables real-time adjustment of the focal plane to track the vessels’ profile. We have experimentally demonstrated that contour scanning improves the signal-to-noise ratio of conventional OR-PAM by as much as 41% and shortens the image acquisition time by 3.2 times. Moreover, contour-scanning OR-PAM more accurately quantifies vessel density and diameter, and has been applied to studying tumors with uneven surfaces

Three-dimensional arbitrary trajectory scanning photoacoustic microscopy

We have enhanced photoacoustic microscopy with three-dimensional arbitrary trajectory (3-DAT) scanning, which can rapidly image selected vessels over a large field of view (FOV) and maintain a high signal-to-noise ratio (SNR) despite the depth variation of the vessels. We showed that hemoglobin oxygen saturation (sO_2) and blood flow can be measured simultaneously in a mouse ear in vivo at a frame rate 67 times greater than that of a traditional two-dimensional raster scan. We also observed sO_2 dynamics in response to switching from systemic hypoxia to hyperoxia

Microvascular quantification based on contour-scanning photoacoustic microscopy

Accurate quantification of microvasculature remains of interest in fundamental pathophysiological studies and clinical trials. Current photoacoustic microscopy can noninvasively quantify properties of the microvasculature, including vessel density and diameter, with a high spatial resolution. However, the depth range of focus (i.e., focal zone) of optical-resolution photoacoustic microscopy (OR-PAM) is often insufficient to encompass the depth variations of features of interest—such as blood vessels—due to uneven tissue surfaces. Thus, time-consuming image acquisitions at multiple different focal planes are required to maintain the region of interest in the focal zone. We have developed continuous three-dimensional motorized contour-scanning OR-PAM, which enables real-time adjustment of the focal plane to track the vessels’ profile. We have experimentally demonstrated that contour scanning improves the signal-to-noise ratio of conventional OR-PAM by as much as 41% and shortens the image acquisition time by 3.2 times. Moreover, contour-scanning OR-PAM more accurately quantifies vessel density and diameter, and has been applied to studying tumors with uneven surfaces

Three-dimensional arbitrary trajectory scanning photoacoustic microscopy

We have enhanced photoacoustic microscopy with three-dimensional arbitrary trajectory (3-DAT) scanning, which can rapidly image selected vessels over a large field of view (FOV) and maintain a high signal-to-noise ratio (SNR) despite the depth variation of the vessels. We showed that hemoglobin oxygen saturation (sO_2) and blood flow can be measured simultaneously in a mouse ear in vivo at a frame rate 67 times greater than that of a traditional two-dimensional raster scan. We also observed sO_2 dynamics in response to switching from systemic hypoxia to hyperoxia

Fully motorized optical-resolution photoacoustic microscopy

We have developed fully motorized optical-resolution photoacoustic microscopy (OR-PAM), which integrates five complementary scanning modes and simultaneously provides a high imaging speed and a wide field of view (FOV) with 2.6 μm lateral resolution. With one-dimensional (1D) motion-mode mechanical scanning, we measured the blood flow through a cross section of a blood vessel in vivo. With two-dimensional (2D) optical scanning at a laser repetition rate of 40 kHz, we achieved a 2 kHz B-scan rate over a range of 50 μm with 20 A-lines and 50 Hz volumetric-scan rate over a FOV of 50  μm×50  μm with 400 A-lines, which enabled real-time tracking of cellular dynamics in vivo. With synchronized 1D optical and 2D mechanical hybrid scanning, we imaged a 10  mm×8  mm FOV within three minutes, which is 20 times faster than the conventional mechanical scan in our second-generation OR-PAM. With three-dimensional mechanical contour scanning, we maintained the optimal signal-to-noise ratio and spatial resolution of OR-PAM while imaging objects with uneven surfaces, which is essential for quantitative studies

Vessel segmentation analysis of ischemic stroke images acquired with photoacoustic microscopy

We have applied optical-resolution photoacoustic microscopy (OR-PAM) for longitudinal monitoring of cerebral metabolism through the intact skull of mice before, during, and up to 72 hours after a 1-hour transient middle cerebral artery occlusion (tMCAO). The high spatial resolution of OR-PAM enabled us to develop vessel segmentation techniques for segment-wise analysis of cerebrovascular responses

Optical-resolution photoacoustic microscopy of ischemic stroke

A major obstacle in understanding the mechanism of ischemic stroke is the lack of a tool to noninvasively or minimally invasively monitor cerebral hemodynamics longitudinally. Here, we applied optical-resolution photoacoustic microscopy (OR-PAM) to longitudinally study ischemic stroke induced brain injury in a mouse model with transient middle cerebral artery occlusion (MCAO). OR-PAM showed that, during MCAO, the average hemoglobin oxygen saturation (sO2) values of feeder arteries and draining veins within the stroke core region dropped ~10% and ~34%, respectively. After reperfusion, arterial sO_2 recovered back to the baseline; however, the venous sO_2 increased above the baseline value by ~7%. Thereafter, venous sO_2 values were close to the arterial sO_2 values, suggesting eventual brain tissue infarction

Optical-resolution photoacoustic microscopy of ischemic stroke

A major obstacle in understanding the mechanism of ischemic stroke is the lack of a tool to noninvasively or minimally invasively monitor cerebral hemodynamics longitudinally. Here, we applied optical-resolution photoacoustic microscopy (OR-PAM) to longitudinally study ischemic stroke induced brain injury in a mouse model with transient middle cerebral artery occlusion (MCAO). OR-PAM showed that, during MCAO, the average hemoglobin oxygen saturation (sO2) values of feeder arteries and draining veins within the stroke core region dropped ~10% and ~34%, respectively. After reperfusion, arterial sO_2 recovered back to the baseline; however, the venous sO_2 increased above the baseline value by ~7%. Thereafter, venous sO_2 values were close to the arterial sO_2 values, suggesting eventual brain tissue infarction

Fully motorized optical-resolution photoacoustic microscopy

Fully motorized optical-resolution photoacoustic microscopy integrates five complementary scanning modes and simultaneously provides a high imaging speed (2 kHz B-scan rate) and a large field of view (10 × 8 mm^2) for fast quantitative imaging

Fully motorized optical-resolution photoacoustic microscopy

We have developed fully motorized optical-resolution photoacoustic microscopy (OR-PAM), which integrates five complementary scanning modes and simultaneously provides a high imaging speed and a wide field of view (FOV) with 2.6 μm lateral resolution. With one-dimensional (1D) motion-mode mechanical scanning, we measured the blood flow through a cross section of a blood vessel in vivo. With two-dimensional (2D) optical scanning at a laser repetition rate of 40 kHz, we achieved a 2 kHz B-scan rate over a range of 50 μm with 20 A-lines and 50 Hz volumetric-scan rate over a FOV of 50  μm×50  μm with 400 A-lines, which enabled real-time tracking of cellular dynamics in vivo. With synchronized 1D optical and 2D mechanical hybrid scanning, we imaged a 10  mm×8  mm FOV within three minutes, which is 20 times faster than the conventional mechanical scan in our second-generation OR-PAM. With three-dimensional mechanical contour scanning, we maintained the optimal signal-to-noise ratio and spatial resolution of OR-PAM while imaging objects with uneven surfaces, which is essential for quantitative studies