Name: a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity
Uploaded: 2017-05-07T13:00:00
Description: Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

This work was supported by a Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant #40352, Campus Alberta for Innovation Program Chair, Alberta Alzheimer Research Program to MHM, and NSERC CREATE in BIF doctoral fellowship and AIHS postgraduate fellowship to MK. We thank Pu Min Wang for the development of this protocol and for surgical training, and Behroo Mirza Agha and Di Shao for husbandry.

The following protocol follows the University of Lethbridge Animal Care Committee (ACC) guidelines, and is conducted in accordance with the standards of the Canadian Council on Animal Care (CCAC).
1. Preparation
<ol>
	<li>For prolonged study periods, autoclave all opened surgical supplies and ensure that sterility is maintained throughout the surgery. If multiple surgeries are required, autoclave between surgeries.&#160;</li>
	<li>Ensure there is plenty of brain buffer on hand (at least 50 m...

<ol>
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This innovative protocol for a large cranial window enables simultaneous imaging over the temporal and parietal areas of the cerebral cortex. Combined with optical imaging, it can help to reveal neural dynamics within cortical areas during spontaneous and stimulus-induced activity. This expansive craniotomy also exposes a large extension of the cortical vasculature network, including the proximal end of the middle cerebral artery (MCA), enabling in vivo imaging of blood flow and direct manipulation of lateral ve...

The craniotomy is a standard procedure used by neuroscientists to reveal a portion of the brain. Since the dawn of electrophysiology, the craniotomy has allowed unprecedented breakthroughs in the field of neuroscience. Dense mapping of the cerebral cortex with electrodes has led to experiments testing hypotheses and theories based on these maps. We have recently entered a new era where the craniotomy is being utilized for in vivo imaging of cortical blood flow1,2,3 and neurovascular architecture4, enabling real time visualization of cortical activity within the exposed areas5,6,7. Although many studies use craniotomies combined with in vivo optical imaging techniques to study the structure and function of cortical neurons, glia, and cortical vasculature8,9, further investigations are limited by small areas of exposed cortex (but see10).
The purpose of this protocol is to provide a method for creating a large lateral craniotomy, exposing the cerebral cortex from the midline to the squamosal bone, and extending beyond bregma and lambda. This large craniotomy enables simultaneous viewing of the association cortices (retrosplenial, cingulate, and parietal), primary and secondary motor, somatosensory, visual, and the auditory cortex. This method has been previously coupled with voltage sensitive dye imaging (VSDI) to investigate how multiple cortical areas interact with one another during spontaneous and stimulus-induced cortical activity5,11,12. The most challenging aspects of this procedure include positioning the head of the animal, fixing the head plate, and avoiding hemorrhage while separating the temporal muscle from the parietal bone. Care must also be taken during the drilling and skull removal processes as the skull curves at an oblique angle.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Heating Pad&#160;</td>
			<td>FHC</td>
			<td>40-90-2</td>
			<td></td>
		</tr>
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			<td>Fine Scissors</td>
			<td>Fine Science Tools</td>
			<td>14058-09</td>
			<td></td>
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		<tr>
			<td>Forceps&#160;</td>
			<td>Fine Science Tools</td>
			<td>11251-35</td>
			<td>2 or more pairs are recommended</td>
		</tr>
		<tr>
			<td>Spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15000-00, 15000-10</td>
			<td>1 pair should be designated for dura removal&#160;</td>
		</tr>
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			<td>Jet tooth shade powder</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Powder</td>
			<td>to be mixed with the Jet Liquid</td>
		</tr>
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			<td>Jet tooth shade liquid</td>
			<td>LANG Dental</td>
			<td>Jet Tooth Shade Liquid</td>
			<td>to be mixed wihth the Jet Powder&#160;</td>
		</tr>
		<tr>
			<td>Drill Heads - Carbide Burs FG 1/4 389</td>
			<td>Midwest Dental</td>
			<td>385201</td>
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			<td>Agarose Powder</td>
			<td>Sigma-Aldrich</td>
			<td>A9793</td>
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			<td>Gelfoam</td>
			<td>Sinclair Dental Canada</td>
			<td>Pfizer Gelfoam</td>
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			<td>Isoflurane</td>
			<td>Western Drug Distribution Centre Ltd</td>
			<td>124125</td>
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			<td>Lidocaine 2% Epinephrine</td>
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			<td>Dexamethazone 5 mg/mL</td>
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</table>

a large lateral craniotomy procedure for mesoscale wide-field optical imaging of brain activity

The craniotomy is a commonly performed procedure to expose the brain for in vivo experiments. In mouse research, most labs utilize a small craniotomy, typically 3 mm x 3 mm. This protocol introduces a method for creating a substantially larger 7 mm x 6 mm cranial window exposing most of a cerebral hemisphere over the mouse temporal and parietal cortices (e.g., bregma 2.5 - 4.5 mm, lateral 0 - 6 mm). To perform this surgery, the head must be tilted approximately 30&#176; and much of the temporal muscle must be retracted. Due to the large amount of bone removal, this procedure is intended only for acute experiments with the animal anesthetized throughout the surgery and experiment.
The main advantage of this innovative large lateral cranial window is to provide simultaneous access to both medial and lateral areas of the cortex. This large unilateral cranial window can be used to study the neural dynamics between cells, as well as between different cortical areas by combining multi-electrode electrophysiological recordings, imaging of neuronal activity (e.g., intrinsic or extrinsic imaging), and optogenetic stimulation. Additionally, this large craniotomy also exposes a large area of cortical blood vessels, allowing for direct manipulation of the lateral cortical vasculature.

To study the interactions between cortical areas within a single hemisphere, we used a large craniotomy extending across the sagittal sinus and 5 - 6 mm lateral. This cranial window included primary (motor, somatosensory, visual, auditory), secondary (motor, visual), and association (retrosplenial, cingulate, parietal association) cortices of right brain hemispheres (Figure 3A). For this work we used voltage sensitive dye (VSD) imaging, which reflects changes...

Watch this Scientific Journal Video about A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity at JoVE.com

A Large Lateral Craniotomy Procedure for Mesoscale Wide-field Optical Imaging of Brain Activity

This protocol presents a method for creating a large unilateral craniotomy over the temporal and parietal regions of the mouse cerebral cortex. This is especially useful for real time imaging over an expansive area of a cortical hemisphere.

The craniotomy is a commonly performed procedure to expose the brain for in vivo experiments. In mouse research, most labs utilize a small craniotomy, ...

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