title: Two-photon microscopy as a tool to study blood flow and neurovascular coupling in the rodent brain.
abstract: The cerebral vascular system services the constant demand for energy during neuronal activity in the brain. Attempts to delineate the logic of neurovascular coupling have been greatly aided by the advent of two-photon laser scanning microscopy to image both blood flow and the activity of individual cells below the surface of the brain. Here we provide a technical guide to imaging cerebral blood flow in rodents. We describe in detail the surgical procedures required to generate cranial windows for optical access to the cortex of both rats and mice and the use of two-photon microscopy to accurately measure blood flow in individual cortical vessels concurrent with local cellular activity. We further provide examples on how these techniques can be applied to the study of local blood flow regulation and vascular pathologies such as small-scale stroke.
authors.affiliation: Department of Physics, University of California at San Diego, La Jolla, CA 92093-0374, USA.
authors.forename: Andy Y
authors.initials: AY
authors.lastname: Shih
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Driscoll
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Drew
authors.forename: Nozomi
authors.initials: N
authors.lastname: Nishimura
authors.forename: Chris B
authors.initials: CB
authors.lastname: Schaffer
authors.forename: David
authors.initials: D
authors.lastname: Kleinfeld
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply*; D002560:Cerebrovascular Circulation / Q000502:physiology*; D006439:Hemodynamics / Q000502:physiology*; D051379:Mice; D008853:Microscopy / Q000379:methods*; D017785:Photons; D051381:Rats