Video-rate volumetric optical coherence tomography (vOCT) is relatively young in the field of OCT imaging but has great potential in biomedical applications. Due to the recent development of the MHz range swept laser sources, vOCT has started to gain attention in the community. Here, we report the first in vivo video-rate volumetric OCT-based microangiography (vOMAG) system by integrating an 18-kHz resonant microelectromechanical system (MEMS) mirror with a 1.6-MHz FDML swept source operating at ∼1.3 μm wavelength. Because the MEMS scanner can offer an effective B-frame rate of 36 kHz, we are able to engineer vOMAG with a video rate up to 25 Hz. This system was utilized for real-time volumetric in vivo visualization of cerebral microvasculature in mice. Moreover, we monitored the blood perfusion dynamics during stimulation within mouse ear in vivo. We also discussed this system’s limitations. Prospective MEMS-enabled OCT probes with a real-time volumetric functional imaging capability can have a significant impact on endoscopic imaging and image-guided surgery applications.
We demonstrate in vivo volumetric optical microangiography at ∼200 volumes/s by the use of 1.6 MHz Fourier domain mode-locking swept source optical coherence tomography and an effective 36 kHz microelectromechanical system (MEMS) scanner. We propose an intervolume analysis strategy to contrast the dynamic blood flow signal from the static tissue background. The proposed system is demonstrated by imaging cerebral blood flow in mice in vivo. For the first time, imaging speed, sensitivity, and temporal resolution become possible for a direct four-dimensional observation of microcirculations within live body parts.
Traumatic brain injury (TBI) is a form of brain injury caused by sudden impact on brain by an external mechanical force.
Following the damage caused at the moment of injury, TBI influences pathophysiology in the brain that takes place
within the minutes or hours involving alterations in the brain tissue morphology, cerebral blood flow (CBF), and
pressure within skull, which become important contributors to morbidity after TBI. While many studies for the TBI
pathophysiology have been investigated with brain cortex, the effect of trauma on intracranial tissues has been poorly
studied. Here, we report use of high-resolution optical microangiography (OMAG) to monitor the changes in cranial
meninges beneath the skull of mouse after TBI. TBI is induced on a brain of anesthetized mouse by thinning the skull
using a soft drill where a series of drilling exert mechanical stress on the brain through the skull, resulting in mild brain
injury. Intracranial OMAG imaging of the injured mouse brain during post-TBI phase shows interesting
pathophysiological findings in the meningeal layers such as widening of subdural space as well as vasodilation of
subarachnoid vessels. These processes are acute and reversible within hours. The results indicate potential of OMAG to
explore mechanism involved following TBI on small animals in vivo.
Fluorescent labeling has opened up the possibility of clarifying the complex distribution and circuit wiring of specific neural circuits for particular functions. To acquire the brain-wide fluorescently labeled neural wiring, we have previously developed the fluorescence micro-optical sectioning tomography imaging system. This employs simultaneous mechanical sectioning and confocal imaging of the slices, and is capable of acquiring the image dataset of a centimeter-sized whole-mouse brain at a voxel resolution of 1 μm. We analyze the key optical considerations for the use of an acousto-optical deflector (AOD) scanner-based confocal detection scheme in this system. As a result, the influence of confocal detection, the imaging site during sectioning, and AOD fast scan mode on signal-to-background noise ratio are described. It is shown that mechanical sectioning to separate the slice and optical sectioning by confocal detection should be combined to maximize background suppression in simultaneous fast scan imaging while sectioning system setup.
Imaging brain circuits is the basis for us to understand brain function and dysfunction. However, imaging axon at micrometer resolution while tracing the centimeter-scale axon projection across the whole-brain is still challenging. Here, we developed a fluorescence micro-optical sectioning tomography (fMOST) imaging system based on confocal fluorescence imaging scheme that can obtain whole brain image stack for visualizing brain circuits at neurite level. We use confocal detection to remove fluorescence background to clearly see one single neurite and use acoustical optical deflector (AOD), an inertia-free beam scanner to realize fast and prolonged stable imaging. We had acquired several complete datasets of whole-mouse brain at a one-micron voxel resolution. Based on these datasets, the uninterrupted tracing of brain-wide, long-distance axonal projections was demonstrated for the first time using a systematic reconstruction and annotation pipeline. Our method is believed to open an avenue to exploring both local and long-distance neural circuits that are related to brain functions and brain diseases down to the neurite level.
A recently reported micro-optical sectioning tomography system has great potential to draw the neuronal circuits of large brain volume with submicron resolution by combining fine mechanic sectioning with simultaneous optical imaging. However, sectioning the fluorescence sample sometimes induces tears between adjacent tiles and causes difficulties in continuous fiber tracing from fluorescence imaging. A confocal detection to recover the interruptions of the nerve fiber is introduced. With a 50-μm-width confocal slit, the signal-to-background ratio is increased 16- to 49-fold more than that without the slit, which effectively improves the detectability of the signal in the interruptions and enables continuous tracing of the neuronal circuits.
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