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      Application of Thinned-Skull Cranial Window to Mouse Cerebral Blood Flow Imaging Using Optical Microangiography

      1 , 1 , 2 , 1 , *
      PLoS ONE
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          In vivo imaging of mouse brain vasculature typically requires applying skull window opening techniques: open-skull cranial window or thinned-skull cranial window. We report non-invasive 3D in vivo cerebral blood flow imaging of C57/BL mouse by the use of ultra-high sensitive optical microangiography (UHS-OMAG) and Doppler optical microangiography (DOMAG) techniques to evaluate two cranial window types based on their procedures and ability to visualize surface pial vessel dynamics. Application of the thinned-skull technique is found to be effective in achieving high quality images for pial vessels for short-term imaging, and has advantages over the open-skull technique in available imaging area, surgical efficiency, and cerebral environment preservation. In summary, thinned-skull cranial window serves as a promising tool in studying hemodynamics in pial microvasculature using OMAG or other OCT blood flow imaging modalities.

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          Most cited references24

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          Optical coherence tomography.

          A technique called optical coherence tomography (OCT) has been developed for noninvasive cross-sectional imaging in biological systems. OCT uses low-coherence interferometry to produce a two-dimensional image of optical scattering from internal tissue microstructures in a way that is analogous to ultrasonic pulse-echo imaging. OCT has longitudinal and lateral spatial resolutions of a few micrometers and can detect reflected signals as small as approximately 10(-10) of the incident optical power. Tomographic imaging is demonstrated in vitro in the peripapillary area of the retina and in the coronary artery, two clinically relevant examples that are representative of transparent and turbid media, respectively.
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            Collateral circulation.

            The collateral circulation plays a pivotal role in the pathophysiology of cerebral ischemia. Current knowledge of the collateral circulation remains sparse, largely because of prior limitations in methods for evaluation of these diminutive routes of cerebral blood flow. Anatomic descriptions of the collateral circulation often focus on more proximal anastomoses at the circle of Willis, neglecting secondary collateral pathways provided by leptomeningeal vessels. Pathophysiological recruitment of collateral vessels likely depends on the temporal course of numerous compensatory hemodynamic, metabolic, and neural mechanisms. Subsequent endurance of these protective vascular pathways may determine the severity of ischemic injury. Characterization of the collateral circulation with advanced neuroimaging modalities that provide angiographic information and perfusion data may elucidate critical determinants of collateral blood flow. Such information on the status of the collateral circulation may be used to guide therapeutic interventions. Prognostication and risk stratification may also be improved by routine evaluation of collateral blood flow. Contemporary understanding of the collateral circulation may be greatly enhanced through further refinement of neuroimaging modalities that correlate angiographic findings with perfusion status, providing the basis for future therapeutic and prognostic applications.
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              Dendritic spines as basic functional units of neuronal integration.

              Most excitatory synaptic connections occur on dendritic spines. Calcium imaging experiments have suggested that spines constitute individual calcium compartments, but recent results have challenged this idea. Using two-photon microscopy to image fluorescence with high resolution in strongly scattering tissue, we measured calcium dynamics in spines from CA1 pyramidal neurons in slices of rat hippocampus. Subthreshold synaptic stimulation and spontaneous synaptic events produced calcium accumulations that were localized to isolated spines, showed stochastic failure, and were abolished by postsynaptic blockers. Single somatic spikes induced fast-peaking calcium accumulation in spines throughout the cell. Pairing of spikes with synaptic stimulation was frequently cooperative, that is, it resulted in supralinear calcium accumulations. We conclude: (1) calcium channels exist in spine heads; (2) action potentials invade the spines; (3) spines are individual calcium compartments; and (4) spines can individually detect the temporal coincidence of pre- and postsynaptic activity, and thus serve as basic functional units of neuronal integration.

                Author and article information

                Role: Editor
                PLoS One
                PLoS ONE
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                26 November 2014
                : 9
                : 11
                : e113658
                [1 ]Department of Bioengineering, University of Washington, Seattle, Washington, United States of America
                [2 ]Department of Electrical Engineering, University of Washington, Seattle, Washington, United States of America
                Singapore Immunology Network, Singapore
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: YL UB. Performed the experiments: YL UB. Analyzed the data: UB. Contributed reagents/materials/analysis tools: RW. Wrote the paper: YL UB RW.

                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                : 16 June 2014
                : 2 October 2014
                Page count
                Pages: 13
                Funding came from the National Institutes of Health (R01EB009682 and R01HL093140). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Research Article
                Biology and Life Sciences
                Biomedical Engineering
                Engineering and Technology
                Medicine and Health Sciences
                Cardiovascular Imaging
                Research and Analysis Methods
                Imaging Techniques
                In Vivo Imaging
                Custom metadata
                The authors confirm that, for approved reasons, some access restrictions apply to the data underlying the findings. Data are available at Figshare: http://figshare.com/s/0d346be0609011e4b05506ec4bbcf141.



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