Name: a craniotomy surgery procedure for chronic brain imaging
Uploaded: 2008-02-15T11:06:00
Description: Watch this Scientific Journal Video about A Craniotomy Surgery Procedure for Chronic Brain Imaging at JoVE.com

<ol>
<li>Anesthetize mice&nbsp;with isoflurane (4% for induction, 1.5-2% for surgery) using IACUC approved procedures. It is important that tail and/or toe pinches are used in order to ensure the animal is fully sedated.</li>
<li>Using a rodent trimmer, shave the hair from the back of the neck up to the eyes.</li>
<li>Place&nbsp;the mouse in a stereotaxic frame, over a surgery water re-circulating blanket. Firmly secure the head with ear bars.</li>
<li>Apply eye ointment, in order to prevent the animal's eye from drying out. </li>
<li>Administe...

<ol>
	<li>Svoboda, K., Denk, W., Kleinfeld, D., Tank, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=+In+vivo+dendritic+calcium+dynamics+in+neocortical+pyramidal+neurons."> In vivo dendritic calcium dynamics in neocortical pyramidal neurons.</a> <em style='font-style: italic'>Nature</em>. <b>85</b>, 161-165 (1997).</li><li>Lendvai, B., Stern, E. A., Chen, B., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+plasticity+of+dendritic+spines+in+the+developing+rat+barrel+cortex+in+vivo.">Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo.</a> <em style='font-style: italic'>Nature</em>. <b>404</b>, 876-881 (2000).</li><li>Trachtenberg, J. T., Chen, B., Knott, G. W., Feng, G., Sanes, J. R., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+in+vivo+imaging+of+experience-dependent+synaptic+plasticity+in+adult+cortex.">Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex.</a> <em style='font-style: italic'>Nature</em>. <b>420</b>, 788-794 (2002).</li><li>Portera-Cailliau, C., Weimer, R. M., DePaola, V., Caroni, P., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diverse+modes+of+axon+elaboration+in+the+developing+neocortex.">Diverse modes of axon elaboration in the developing neocortex.</a> <em style='font-style: italic'>PLoS Biol</em>. <b>3</b>, (2005).</li><li>Holtmaat, A., Wilbrecht, L., Knott, G. W., Welker, E., Svoboda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experience-dependent+and+cell-type-specific+spine+growth+in+the+neocortex.">Experience-dependent and cell-type-specific spine growth in the neocortex.</a> <em style='font-style: italic'>Nature</em>. <b>441</b>, 979-983 (2006).</li></ol>

<p class="jove_content">As we have shown in the video and in the supplementary figures, the cranial window preparation, combined with the use of two-photon microscopy, is a very powerful tool to study in vivo the structure and function of the neocortex. The technique requires rigorous training to become familiar with the relevant anatomy and the fine surgical procedures and skills that this preparation requires.  Only pristine surgeries can be used for chronic imaging. If the dura is manipulated excessively or punctured, the preparation should ...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Carprofen (Rimadyl)</td>
<td>Drug</td>
<td>Pfizer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane (Aerrane)</td>
<td>Surgery</td>
<td>Baxter</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Dexamethasone</td>
<td>Drug</td>
<td>Baxter</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ortho-Jet Powder</td>
<td>Reagent</td>
<td>LANG</td>
<td>&nbsp;</td>
<td>To be mixed with the acrylic</td>
</tr>
<tr>
<td>Jet-Acrylic Liquid</td>
<td>Reagent</td>
<td>LANG</td>
<td>&nbsp;</td>
<td>To be mixed with Ortho-Jet Powder</td>
</tr>
<tr>
<td>Round Glass Cover Slip</td>
<td>Tool</td>
<td>Electron Microscopy Sciences</td>
<td>72195-05</td>
<td>5 mm diameter</td>
</tr>
<tr>
<td>Gelfoam</td>
<td>Surgery</td>
<td>Pharmacia & Upjohn Co. (Pfizer)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
			<div>
			Titanium bars are custom-made		</div>
		

a craniotomy surgery procedure for chronic brain imaging

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an extremely useful method, because it allows the study of the live intact brain. With appropriate preparations, this technique allows the observation of the same cortical area chronically, from minutes to months. In this video, we show a preparation for chronic in vivo imaging of the brain using two-photon microscopy. This technique was initially pioneered by Dr. Karel Svoboda, who is now a Howard Hughes Medical Institute Investigator at Janelia Farm. Preparations like the one shown here can be used for imaging of neocortical structure (e.g., dendritic and axonal dynamics),  to record neuronal activity using calcium-sensitive dyes, to image cortical blood flow dynamics, or for intrinsic optical imaging studies. Deep imaging of the neocortex is possible with optimal cranial window surgeries. Operating under the most sterile conditions possible to avoid infections, together with using extreme care to do not damage the dura mater during the surgery, will result in successful and long-lasting glass-covered cranial windows.


Watch this Scientific Journal Video about A Craniotomy Surgery Procedure for Chronic Brain Imaging at JoVE.com

A Craniotomy Surgery Procedure for Chronic Brain Imaging

<p class="jove_content">This video and protocol demonstrate how to implant a glass-covered cranial window in rodents. These preparations can be used for chronic in vivo two-photon imaging of the neocortex over time scales of months. It may also be used for other types of imaging, including optical intrinsic signal imaging.</p>

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an ...

Research

JoVE Journal

Biology

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

Micro-drive Array for Chronic <em>in vivo</em> Recording: Drive Fabrication

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. ...

Micro-drive Array for Chronic <em>in vivo</em> Recording: Tetrode Assembly

Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential ...

Murine Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural ...

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol ...

Mapping the After-effects of Theta Burst Stimulation on the Human Auditory Cortex with Functional Imaging

Kortlægning af eftervirkningerne af Theta brast Stimulation om den menneskelige auditive cortex med Functional Imaging

Auditory cortex pertains to the processing of sound, which is at the basis of speech or music-related processing1. However, despite considerable recent ...

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

Isolering af humane hepatocytter ved en to-trins collagenaseperfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

Undersøgelse af protein-protein interaktioner i levende celler Brug Bioluminescens Resonance Energy Transfer

Assays based on Bioluminescence Resonance Energy Transfer (BRET) provide a sensitive and reliable means to monitor protein-protein interactions in live ...