1. Preparation
1.1. Anesthesia of the rats
<ol>
	<li>Place the rat into a chamber prefilled with isoflurane (depending on the size of the chamber about 3 minutes of 5%) to anesthetize the rat.</li>
	<li>Once anesthetized, remove the rat and weigh it.</li>
	<li>Place the nose of the rat into the anesthesia mask (isoflurane 2-3% in oxygen 1 L/min) to maintain anesthesia. During anesthesia with isoflurane, the heart rates of rats decrease to 300-340 bpm (380-420 bpm in mice).</li>
</ol>
1.2. Insert a tail vein cannula
<ol>
	<li>Wash the tail of the rat with warm water (38 &deg;C) and soap to allow the blood vessels to be more easily seen.</li>
	<li>Cannulate either vein, which can be easily seen on the lateral aspect of the tail. Using a 22-24 G i.v. catheter, begin attempts approximately 1/3 of the way from the tip.</li>
	<li>If the initial efforts are unsuccessful, retry more proximally.</li>
	<li>Take blood sample for analysis of hematocrit (necessary to calculate extracellular volume fraction= ECV).</li>
	<li>Secure the i.v. catheter with skin glue and tape.</li>
</ol>
1.3. Positioning of the electrodes
<ol>
	<li>Clean all footpads with alcohol saturated swab or with soap and water.</li>
	<li>Four electrodes are needed, one for each foot.</li>
	<li>Attach the electrodes to the middle of the footpad and secure them with adhesive tape.</li>
	<li>Twist or plait the ECG cables around each other to minimize the formation of loops that would cause interference with the ECG signal.</li>
</ol>
2. Scanning
2.1. Positioning of the animal and monitoring
<ol>
	<li>Place the rat in a supine or prone position onto the imaging tube and position the heart in the center of the coil.</li>
	<li>Arrange the rat so that its nose rests in the cone which delivers isoflurane anesthesia (2-3% in oxygen 1 L/min). The rat is allowed to breath freely throughout the entire imaging procedure.</li>
	<li>Place the sensor of the thermometer at the anal orifice and fix it on the table with tape.</li>
	<li>Body temperature is maintained at 38&plusmn;1 &deg;C by a warm water heating system. The warming mat is placed under the lower aspect of the animal which extends outside the coil. Please see the schematic representation of the MRI experimental setup (Figure 2) for more details.</li>
	<li>Connect the ECG electrodes, once the ECG signal is stable begin imaging.</li>
</ol>
2.2. Scan protocol/ Imaging parameters
<ol>
	<li>In this study a Philips Ingenia 3.0T clinical MRI system with a solenoid coil with 70 mm inner diameter was used [MR system requirements are a small bore (e.g. 16 cm) or large bore (human size, e.g. 65 cm) with dedicated small animal coil, ECG gating capabilities for heart rates up to 500/min, oxygen supply and a removal system for anesthetic gases].</li>
	<li>Start with a survey to locate the heart in the center of the MRI field of view.</li>
	<li>Once the position is correct, continue with two-chamber and four-chamber cine MRI views to determine the geometry of the stack of short axis images.</li>
	<li>For SALLI, position mid-cavity short axis slice on the two- and four-chamber images (PreSALLI). A combination of 4,000 msec acquisition duration and 4,000 msec relaxation duration allows for sufficient recovery of magnetization along the z-axis and prevents overheating of gradients. To enable functional analysis, phases should be set to at least 12, with temporal undersampling of factor 2 to accelerate acquisition.</li>
	<li>Inject the gadolinium-based contrast agent into the indwelling catheter, without moving the rat. Wait five minutes.</li>
	<li>Start planning SALLI as multislice short axis stack, consisting of at least 7 slices (slice thickness 2.4 mm). Position the slices so that the most distal is below the apex of the heart and the top level is just distal to the cardiac valves. Try to ensure that the short axes are perpendicular to the septum. Almost every 30 minutes (around three SALLIs) inject another dose of contrast agent.</li>
	<li>When image acquisition is finished remove the rat from the scanner and allow it to recover from anesthesia.</li>
</ol>
3. Image Reconstruction
Image reconstruction may be performed either online using predefined reconstruction parameters, or offline using a dedicated image reconstruction tool:
3.1. Online Reconstruction
<ol>
	<li>Various Parameters can be defined including:</li>
</ol>
<ul>
	<li>For T1 mapping: location and width of the reconstruction window within the cardiac cycle (e.g. from 50 to 100 msec to achieve systolic T1 map at heart rate 300/min)</li>
	<li>For IR-prepared (LGE) images: location and width of the inversion time (TI) window and number of images to be reconstructed within that window (e.g. TI 100-300 msec, 5 images, to generate 5 LGE images with TIs 100-150-200-250-300 msec).</li>
</ul>
3.2. Offline Reconstruction
<ol>
	<li>When using a dedicated software package (Gyrotools, Z&uuml;rich, Switzerland) for image reconstruction, extract the three sets of data (cine MR, inversion recovery and T1 maps) in a more interactive way.</li>
	<li>Adjust the definition of end-systole or end-diastole for the T1 by selecting the corresponding cine image.</li>
	<li>Visualize T1 maps as color maps. Find exact values by passing the mouse over an area of interest. Images can be stored in various image formats for further analysis (e.g. calculation of ECV from pre- and post-contrast T1 values of myocardium and blood pool) 12.</li>
</ol>

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All animal procedures were conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals, and approved by the local animal care authorities. No conflicts of interest declared.

Here we present a method to generate multi-modality (cine MR, inversion-recovery and T1 maps) MR images in small animals. Small animal research plays an increasingly important role in cardiovascular disease research, and CMR is a powerful tool allowing us to study the function, structure and tissue composition of the myocardium. However, small animal CMR has a number of unique challenges owing to the high heart rates and small heart size. Improved methods of imaging, including more time efficient methods will play a furt...

Myocarditis is a major cause of acute heart failure, sudden death, and chronic dilated cardiomyopathy 7. CMR has been established as the gold standard technique for measurement of function and for in vivo tissue analysis. Novel imaging techniques and improvements in imaging could not only improve the diagnosis of myocarditis, but also aid the study of the pathophysiology and speed identification of therapeutic targets 8-10. Small animal imaging is an important tool for the study of cardiovascular diseases. Techniques routinely used in clinical CMR, such as late gadolinium enhancement (LGE) cannot easily be transferred to small animal CMR due to the animals' high heart rates; but could already be shown 11. The small animal Look-Locker inversion recovery (SALLI) approach generates multi-modal image data sets allowing for the comprehensive assessment of both cardiac function and morphology (Figures 1 and 3). Here we show, in detail, the procedure and setup for small animal imaging with a typical SALLI protocol. In particular we show the reconstruction and analysis of the T1 imaging data sets.

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>Adhesive Tape (Silkafix)</td>
			<td>Lohmann und Rauscher</td>
			<td>34327</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Gadopentetat-Dimeglumin (Magnevist)</td>
			<td>Bayer</td>
			<td>G-00012163</td>
			<td>2mmol/Kg</td>
		 </tr>
		 <tr>
			<td>Introcan Safety-W (G24)</td>
			<td>B. Braun</td>
			<td>4254503-01</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Red Dot, Neonatal Monitoring Electrode with Pre-Attached Lead Wire</td>
			<td>3M</td>
			<td>2269T</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Skin glue (Histoacryl)</td>
			<td>B. Braun</td>
			<td>1050052</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Scales (Typ 440)</td>
			<td>Kern</td>
			<td>95088</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Skin desinfection (Softasept N)</td>
			<td>B. Braun Petzold</td>
			<td>360250</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Thermometer </td>
			<td>LumaSense Technologies</td>
			<td>Luxtron 812</td>
			<td></td>
		 </tr>
		</tbody>
 </table>

assessment of cardiac function and myocardial morphology using small animal look-locker inversion recovery (salli) mri in rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Here we demonstrate the findings from a healthy Sprague Dawley Rat. With a well-prepared animal setup we described before, it is possible to get a stable ECG signal. Similar to human cardiac MRI protocols, we start with a survey to locate the heart. If the position of the rat in the coil is correct, we continue with a two-chamber view and a four-chamber view to detect the geometry for the SALLI stack.
The SALLI technique produces three types of images acquired at the same time. These are, firs...

Watch this Scientific Journal Video about Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats at JoVE.com

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

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12.6K Views.  German Heart Institute Berlin. Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.