SignificancePerturbations in the microcirculatory system have been observed in neurological conditions, such as Alzheimer’s disease or systemic inflammation. However, changes occurring at the level of the capillary are difficult to translate to biomarkers that could be measured macroscopically.AimWe aim to evaluate whether transit time changes reflect capillary stalling and to what degree.ApproachWe employ a combined spectral optical coherence tomography (OCT) and fluorescence optical imaging (FOI) system to investigate the relation between capillary stalling and transit time in a mouse model of systemic inflammation induced by intraperitoneal injection of lipopolysaccharide. Angiograms are obtained using OCT, and fluorescence signal images are acquired by the FOI system upon intravenous injection of fluorescein isothiocyanate via a catheter inserted into the tail vein.ResultsOur findings reveal that lipopolysaccharide (LPS) administration significantly increases both the percentage and duration of capillary stalling compared to mice receiving a 0.9% saline injection. Moreover, LPS-induced mice exhibit significantly prolonged arteriovenous transit time compared to control mice.ConclusionsThese observations suggest that capillary stalling, induced by inflammation, modulates cerebral mean transit time, a measure that has translational potential.
The cerebral microcirculation plays a crucial role in maintaining cerebral homeostasis and facilitating optimal neuronal activity. Perturbations in this intricate microcirculatory system have been observed in neurological conditions such as Alzheimer's disease or systemic inflammation. However, changes occurring at the level of the capillary are difficult to translate to biomarkers that could be measured macroscopically. In this study, we employed a combined spectral optical coherence tomography (OCT) and intrinsic signal optical imaging (ISOI) system to investigate the capillary stalling and transit time of cerebral blood vessels in a mouse model of systemic inflammation induced by the intraperitoneal injection of lipopolysaccharide (LPS). Our findings reveal that LPS administration significantly increases both the percentage and duration of capillary stalling compared to mice receiving 0.9% saline injection. Moreover, LPS-induced mice exhibit significantly prolonged transit time of cerebral blood vessels compared to control mice. These observations suggest that capillary stalling, induced by inflammation, modulate transit time, a measure that has translational potential.
Recent advancements in sources and detectors operating in the NIR-II wavelengths have driven the emergence of NIR-II intrinsic microscopy. These significant technological strides were necessary because longer wavelengths are known to experience reduced scattering and absorption in biological tissue. Leveraging this optical advantage, the application of the NIR-II spectral domain in microscopy holds the potential to improve the depth of imaging and preserve coherence depth. In this study, we showcase the integration of phase-contrast imaging into a NIR-II reflectance confocal microscope for cortical imaging. By capturing images of cortical cell bodies at depths of up to 800 μm, we demonstrate that the implementation of phase contrast provides clear delineation of cortical cell edges, including myelinated axons, blood vessels, and cortical cell bodies. Additionally, we devised a computational method to enhance dynamic components and generate a digitized vascular network from the acquired images.
SignificanceThe imaging depth of microscopy techniques is limited by the ability of light to penetrate biological tissue. Recent research has addressed this limitation by combining a reflectance confocal microscope with the NIR-II (or shortwave infrared) spectrum. This approach offers significant imaging depth, is straightforward in design, and remains cost-effective. However, the imaging system, which relies on intrinsic signals, could benefit from adjustments in its optical design and post-processing methods to differentiate cortical cells, such as neurons and small blood vessels.AimWe implemented a phase contrast detection scheme to a reflectance confocal microscope using NIR-II spectral range as illumination.ApproachWe analyzed the features retrieved in the images while testing the imaging depth. Moreover, we introduce an acquisition method for distinguishing dynamic signals from the background, allowing the creation of vascular maps similar to those produced by optical coherence tomography.ResultsThe phase contrast implementation is successful to retrieve deep images in the cortex up to 800 μm using a cranial window. Vascular maps were retrieved at similar cortical depth and the possibility of combining multiple images can provide a vessel network.ConclusionsPhase contrast reflectance confocal microscopy can improve the outlining of cortical cell bodies. With the presented framework, angiograms can be retrieved from the dynamic signal in the biological tissue. Our work presents an optical implementation and analysis techniques from a former microscope design.
In this work, we demonstrate the application of Rose Bengal as a photo-thrombotic agent using 2-photon and 3-photon excitation by measuring the response to multiphotonic excitation. We also demonstrate, in the case of 2-photon excitation, that the mechanism used is different from laser injury alone with a control group of FitC injected mice. Preliminary results show that a capillary photo-thrombosis could be performed up to 200 μm and OCT imaging could confirm blockage.
To obtain an accurate representation of a brain structural connectivity, diffusion MRI and fiber tracking depend on a good understanding of white matter fiber structures. Although the tracking methods work well when performed in single orientation fiber bundles, most methods are limited in more complex cases, especially to take into account crossing, fanning, and kissing fibers. A recent international fiber tracking challenge concluded that most tracking algorithms generated 4–5 times more false positive tracks than true tracks on average. This was attributed in large part to a lack of knowledge about the fiber crossing geometry. There is thus a dire need to study more complex fiber geometries to improve the tractography algorithms, for example by classifying those geometries into characteristic crossing topologies (e.g., fanning, curving, bottleneck, pure crossing, ...). Here, we propose a multimodal neuroimaging pipeline to identify and acquire fiber crossing areas in whole mouse brains. Our method uses the Allen Mouse Brain connectivity atlas and tractogram analysis using diffusion MRI techniques to identify candidate regions of interests containing fiber crossings based on two predetermined retrograde viral injection site locations. Based on serial OCT acquisitions, we confirmed the location of crossings. Further experiments will validate in detail the structural nature of crossings using retrograde injections of fluorescent tracers and whole mouse brain serial blockface histology. We believe that this new methodological approach will provide indispensable data for the development of a new generation of tractography algorithms that better resolve complex fiber geometries.
An automated dual-resolution serial optical coherence tomography (2R-SOCT) scanner is developed. The serial histology system combines a low-resolution (25 μm / voxel) 3 × OCT with a high-resolution (1.5 μm / voxel) 40 × OCT to acquire whole mouse brains at low resolution and to target specific regions of interest (ROIs) at high resolution. The 40 × ROIs positions are selected either manually by the microscope operator or using an automated ROI positioning selection algorithm. Additionally, a multimodal and multiresolution registration pipeline is developed in order to align the 2R-SOCT data onto diffusion MRI (dMRI) data acquired in the same ex vivo mouse brains prior to automated histology. Using this imaging system, 3 whole mouse brains are imaged, and 250 high-resolution 40 × three-dimensional ROIs are acquired. The capability of this system to perform multimodal imaging studies is demonstrated by labeling the ROIs using a mouse brain atlas and by categorizing the ROIs based on their associated dMRI measures. This reveals a good correspondence of the tissue microstructure imaged by the high-resolution OCT with various dMRI measures such as fractional anisotropy, number of fiber orientations, apparent fiber density, orientation dispersion, and intracellular volume fraction.
Given known correlations between vascular health and cognitive impairment, the development of tools to image microvasculature in the whole brain could help investigate these correlations. We explore the feasibility of using an automated serial two-photon microscope to image fluorescent gelatin-filled whole rodent brains in three-dimensions (3-D) with the goal of carrying group studies. Vascular density (VD) was computed using automatic segmentation combined with coregistration techniques to build a group-level vascular metric in the whole brain. Focusing on the medial prefrontal cortex, cerebral cortex, the olfactory bulb, and the hippocampal formation, we compared the VD of three age groups (2-, 4.5-, and 8-months-old), for both wild type mice and a transgenic model (APP/PS1) with pathology resembling Alzheimer’s disease (AD). We report a general loss of VD caused by the aging process with a small VD increase in the diseased animals in the somatomotor and somatosensory cortical regions and the olfactory bulb, partly supported by MRI perfusion data. This study supports previous observations that AD transgenic mice show a higher VD in specific regions compared with WT mice during the early and late stages of the disease (4.5 to 8 months), extending results to whole brain mapping.
In this study, an automated serial two-photon microscope was used to image a fluorescent gelatin filled rodent’s brain in 3D. A method to compute vascular density using automatic segmentation was combined with coregistration techniques to build group-level vasculature metrics. By studying the medial prefrontal cortex and the hippocampal formation of 3 age groups (2, 4.5 and 8 months old), we compared vascular density for both WT and an Alzheimer model transgenic brain (APP/PS1). We observe a loss of vascular density caused by the ageing process and we propose further analysis to confirm our results.
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