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Acute two-photon imaging of the neurovascular unit in the cortex of active mice

In vivo two-photon scanning fluorescence imaging is a powerful technique to observe physiological processes from the millimeter to the micron scale in the intact animal. In neuroscience research, a common approach is to install an acute cranial window and head bar to explore neocortical function und...

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Autores principales: Tran, Cam Ha T., Gordon, Grant R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4318346/
https://www.ncbi.nlm.nih.gov/pubmed/25698926
http://dx.doi.org/10.3389/fncel.2015.00011
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author Tran, Cam Ha T.
Gordon, Grant R.
author_facet Tran, Cam Ha T.
Gordon, Grant R.
author_sort Tran, Cam Ha T.
collection PubMed
description In vivo two-photon scanning fluorescence imaging is a powerful technique to observe physiological processes from the millimeter to the micron scale in the intact animal. In neuroscience research, a common approach is to install an acute cranial window and head bar to explore neocortical function under anesthesia before inflammation peaks from the surgery. However, there are few detailed acute protocols for head-restrained and fully awake animal imaging of the neurovascular unit during activity. This is because acutely performed awake experiments are typically untenable when the animal is naïve to the imaging apparatus. Here we detail a method that achieves acute, deep-tissue two-photon imaging of neocortical astrocytes and microvasculature in behaving mice. A week prior to experimentation, implantation of the head bar alone allows mice to train for head-immobilization on an easy-to-learn air-supported ball treadmill. Following just two brief familiarization sessions to the treadmill on separate days, an acute cranial window can subsequently be installed for immediate imaging. We demonstrate how running and whisking data can be captured simultaneously with two-photon fluorescence signals with acceptable movement artifacts during active motion. We also show possible applications of this technique by (1) monitoring dynamic changes to microvascular diameter and red blood cells in response to vibrissa sensory stimulation, (2) examining responses of the cerebral microcirculation to the systemic delivery of pharmacological agents using a tail artery cannula during awake imaging, and (3) measuring Ca(2+) signals from synthetic and genetically encoded Ca(2+) indicators in astrocytes. This method will facilitate acute two-photon fluorescence imaging in awake, active mice and help link cellular events within the neurovascular unit to behavior.
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spelling pubmed-43183462015-02-19 Acute two-photon imaging of the neurovascular unit in the cortex of active mice Tran, Cam Ha T. Gordon, Grant R. Front Cell Neurosci Neuroscience In vivo two-photon scanning fluorescence imaging is a powerful technique to observe physiological processes from the millimeter to the micron scale in the intact animal. In neuroscience research, a common approach is to install an acute cranial window and head bar to explore neocortical function under anesthesia before inflammation peaks from the surgery. However, there are few detailed acute protocols for head-restrained and fully awake animal imaging of the neurovascular unit during activity. This is because acutely performed awake experiments are typically untenable when the animal is naïve to the imaging apparatus. Here we detail a method that achieves acute, deep-tissue two-photon imaging of neocortical astrocytes and microvasculature in behaving mice. A week prior to experimentation, implantation of the head bar alone allows mice to train for head-immobilization on an easy-to-learn air-supported ball treadmill. Following just two brief familiarization sessions to the treadmill on separate days, an acute cranial window can subsequently be installed for immediate imaging. We demonstrate how running and whisking data can be captured simultaneously with two-photon fluorescence signals with acceptable movement artifacts during active motion. We also show possible applications of this technique by (1) monitoring dynamic changes to microvascular diameter and red blood cells in response to vibrissa sensory stimulation, (2) examining responses of the cerebral microcirculation to the systemic delivery of pharmacological agents using a tail artery cannula during awake imaging, and (3) measuring Ca(2+) signals from synthetic and genetically encoded Ca(2+) indicators in astrocytes. This method will facilitate acute two-photon fluorescence imaging in awake, active mice and help link cellular events within the neurovascular unit to behavior. Frontiers Media S.A. 2015-02-05 /pmc/articles/PMC4318346/ /pubmed/25698926 http://dx.doi.org/10.3389/fncel.2015.00011 Text en Copyright © 2015 Tran and Gordon. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Tran, Cam Ha T.
Gordon, Grant R.
Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title_full Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title_fullStr Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title_full_unstemmed Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title_short Acute two-photon imaging of the neurovascular unit in the cortex of active mice
title_sort acute two-photon imaging of the neurovascular unit in the cortex of active mice
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4318346/
https://www.ncbi.nlm.nih.gov/pubmed/25698926
http://dx.doi.org/10.3389/fncel.2015.00011
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