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Two-Photon Imaging of Cerebral Vasodynamics in Awake Mice During Health and Disease

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Advances in Intravital Microscopy

Abstract

The energy demands of the brain are supplied by a dynamic and topologically organized vascular system. In vivo two-photon laser-scanning microscopy has become an essential tool for studying several aspects of cerebrovascular function, including neurovascular coupling, collateral arterial flow, and vasculopathies associated with stroke and neurodegeneration. Recently there has been a strong effort to perform imaging studies on mice in the awake state. This is to avoid the dampening effect that anesthetics have on neurovascular dynamics, and also to enable studies on trained behaviors. Studying awake mice has revealed a rich dynamism of cerebral blood flow control, but also raised a number of challenges in the collection of rigorous and meaningful datasets. In this chapter, we describe techniques routinely used for two-photon blood flow imaging in awake mice. This includes chronic window implantations and head fixation techniques to reduce movement during imaging, as well as blood flow data processing techniques that are robust to movement artifacts. Finally, we describe a non-invasive photothrombotic stroke model that can be performed on awake mice for imaging of ischemic pathology.

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Abbreviations

ACSF:

Artificial cerebral spinal fluid

BOLD fMRI:

Blood oxygen level dependent functional magnetic resonance imaging

PoRTS:

Polished and reinforced thinned skull

RBC:

Red blood cell

TPLSM:

Two-photon laser scanning microscopy

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Acknowledgements

Our work is generously supported by grants to A.Y.S. from the National Institutes of Health for generous support (1R21NS085402-01), Dana Foundation (David Mahoney Neuroimaging Program) and South Carolina Clinical and Translational Institute (UL1TR000062).

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Authors and Affiliations

  1. Department of Neurosciences, Medical University of South Carolina, Charleston, SC, USA

    Philipp M. Summers, Zachary J. Taylor & Andy Y. Shih

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  1. Philipp M. Summers
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  2. Zachary J. Taylor
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  3. Andy Y. Shih
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Correspondence to Andy Y. Shih .

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Editors and Affiliations

  1. National Institutes of Health, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA

    Roberto Weigert

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Summers, P.M., Taylor, Z.J., Shih, A.Y. (2014). Two-Photon Imaging of Cerebral Vasodynamics in Awake Mice During Health and Disease. In: Weigert, R. (eds) Advances in Intravital Microscopy. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-9361-2_2

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