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Fluorescence Microscopy Journal Club

In vivo imaging and analysis of cerebrovascular hemodynamic responses and tissue oxygenation in the mouse brain

by Kassandra Kisler, Divna Lazic, Melanie D Sweeney, Shane Plunkett, Mirna El Khatib, Sergei A Vinogradov, David A Boas, Sava Sakadžic´& Berislav V Zlokovic

Nature Protocols, 2018, 13, pp. 1377 – 1402

Imaging cerebrovascular function in healthy and diseased brains in any of the existing mouse models of neurological and vascular disorders is challenging, especially as methods are often not described in sufficient detail and may vary between laboratories. The authors describe in unprecedented detail, the techniques used to evaluate vascular effects in the mouse brain in such a way that they can be more-easily reproduced in any laboratory with the appropriate instrumentation.

All of the protocols can be performed using any of the current Bruker range of multiphoton microscopes. The in vivo protocols include the use of high-resolution two-photon microscopy to evaluate capillary and arteriolar responses to a stimulus, regional hemodynamic responses, and oxygen delivery to the brain. The paper also describes in step-by-step detail how to measure intrinsic nicotinamidedinucleotide (NADH) fluorescence to understand how blood oxygen supply meets the metabolic demands of activated brain tissue, as well as how to perform resting-state absolute oxygen partial pressure (pO2) measurements of brain tissue using phosphorescence lifetime imaging microscopy (PLIM).