Name: semi-automated optical heartbeat analysis of small hearts
Uploaded: 2009-09-16T13:00:00
Description: Watch this Scientific Journal Video about Semi-automated Optical Heartbeat Analysis of Small Hearts at JoVE.com

<p class="jove_content">KO and AC are supported by a grant and a fellowship from the American Heart Association. SIB and RB are supported by grants from NIH.</p>

<p class="jove_content"><a href="https://www.jove.com/files/ftp_upload/1435/SOHA_overview.pdf">Click here to view an overview of the semi automated heartbeat analysis (SOHA) procedure.</a></p>
<p class="jove_content">Semi-automated optical heartbeat analysis</p>
<p class="jove_title">1. Preprocess -</p>
<p class="jove_content">This step provides information on the diastolic and systolic heart diameters.</p>
<p class="jove_content">Heart edges are identified and marked during maximum diastole and systole. Optical recordings can be advanced at sl...

<ol>
	<li>Cammarato, A., Dambacher, C. M., Reedy, M. C., Knowles, A. F., Kronert, W. A., Bodmer, R., Ocorr, K., Bernstein, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myosin+Transducer+mutations+differentially+affect+motor+function,+myofibril+structure,+and+the+performance+of+skeletal+and+cardiac+muscles.">Myosin Transducer mutations differentially affect motor function, myofibril structure, and the performance of skeletal and cardiac muscles.</a> <em style='font-style: italic'>Mol Biol Cell</em>. <b>19</b> (2), 553-562 (2008).</li><li>Ocorr, K., Reeves, N., Wessells, R. J., Fink, M., Chen, H. -. S. V., Akasaka, T., Yasuda, S., Metzger, J., Giles, W., Posakony, J. W., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=KCNQ+potassium+channel+mutations+cause+cardiac+arrhythmias+in+Drosophila+that+mimic+the+effects+of+aging.">KCNQ potassium channel mutations cause cardiac arrhythmias in Drosophila that mimic the effects of aging.</a> <em style='font-style: italic'>Proc Natl Acad Sci U S A</em>. <b>104</b>, 3943-3948 (2007).</li><li>Fink, M., Callol-Massot, C., Chu, A., Ruiz-Lozano, P., Izpisua Belmonte, J. C., Giles, W., Bodmer, R., Ocorr, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+for+the+detection+and+quantification+of+heartbeat+parameters+in+Drosophila,+zebrafish+and+embryonic+mouse+hearts.">A new method for the detection and quantification of heartbeat parameters in Drosophila, zebrafish and embryonic mouse hearts.</a> <em style='font-style: italic'>Biotechniques</em>. <b>46</b>, 101-113 (2009).</li></ol>

<p class="jove_content">The Drosophila model has proven to be a powerful genetic tool that has been used to address a variety of scientific questions ranging from embryological development to learning and memory. Recently this versatile model organism has been used to investigate the genetics of heart function.  A number of attempts to quantify heart physiology in adult Drosophila have relied on observations made in intact flies through the abdominal cuticle. Most of these approaches depend upon visual observation or recordings of changes in li...

<!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>MatLab software</td>
<td>&nbsp;</td>
<td>Math Works, Inc</td>
<td>&nbsp;</td>
<td>Required environment for the analysis software.</td>
</tr>
<tr>
<td>EM-CCD digital camera</td>
<td>&nbsp;</td>
<td>Hamamatsu Corp.</td>
<td>9100 or 9300</td>
<td>Other high speed digital cameras will also work.</td>
</tr>
<tr>
<td>HC image  data capture software</td>
<td>&nbsp;</td>
<td>Hamamatsu Corp.</td>
<td>&nbsp;</td>
<td>Other image capture software that produces movies in avi format will also work.</td>
</tr>
<tr>
<td>Light Microscope with 10x objective </td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>A dipping lens that eliminates the air water interface greatly improves resolution</td>
</tr>		</tbody>
		</table>
		</div>
	

semi-automated optical heartbeat analysis of small hearts

We have developed a method for analyzing high speed optical recordings from <em>Drosophila</em>, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our Semi-automatic Optical Heartbeat Analysis (SOHA) uses a novel movement detection algorithm that is able to detect cardiac movements associated with individual contractile and relaxation events. The program provides a host of physiologically relevant readouts including systolic and diastolic intervals, heart rate, as well as qualitative and quantitative measures of heartbeat arrhythmicity. The program also calculates heart diameter measurements during both diastole and systole from which fractional shortening and fractional area changes are calculated. Output is provided as a digital file compatible with most spreadsheet programs. Measurements are made for every heartbeat in a record increasing the statistical power of the output. We demonstrate each of the steps where user input is required and show the application of our methodology to the analysis of heart function in all three genetically tractable heart models.


Watch this Scientific Journal Video about Semi-automated Optical Heartbeat Analysis of Small Hearts at JoVE.com

Semi-automated Optical Heartbeat Analysis of Small Hearts

<p class="jove_content">We have developed a Semi-automated Optical Heartbeat Analysis method (SOHA) for analyzing high speed optical recordings from <em>Drosophila</em>, zebrafish and embryonic mouse hearts. We demonstrate the application of our methodology to the analysis of heart function in fruit fly and embryonic mouse hearts.</p>

We have developed a method for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our ...

Research

JoVE Journal

Biology

Proper Care and Cleaning of the Microscope

Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a ...

Major Components of the Light Microscope

The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for ...

Phase Contrast and Differential Interference Contrast (DIC) Microscopy

Phase Contrast and Differential Interference Contrast DIC Microscopy

Phase-contrast microscopy is often used to produce contrast for transparent, non light-absorbing, biological specimens. The technique was discovered by ...

Optical Mapping of Langendorff-perfused Rat Hearts

Optical mapping of the cardiac surface with voltage-sensitive fluorescent dyes has become an important tool to investigate electrical excitation in ...

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing ...

Substrate Generation for Endonucleases of CRISPR/Cas Systems

The  interaction of viruses and their prokaryotic hosts shaped the evolution of  bacterial and archaeal life. Prokaryotes developed several strategies to ...

LeafJ: An ImageJ Plugin for Semi-automated Leaf Shape Measurement

High throughput phenotyping (phenomics) is a powerful tool for linking genes to their functions (see review1 and recent examples2-4). Leaves are the ...

Semi-automated Imaging of Tissue-specific Fluorescence in Zebrafish Embryos

Zebrafish embryos are a powerful tool for large-scale screening of small molecules. Transgenic zebrafish that express fluorescent reporter proteins are ...

Recovery of Adult Zebrafish Hearts for High-throughput Applications

Use of the zebrafish model system for studying development, regeneration, and disease is expanding toward use of adult hearts for cell dissociation and ...

Isolation of Cardiomyocytes from Fixed Hearts for Immunocytochemistry and Ploidy Analysis

The adult mammalian heart is composed of various cell types including cardiomyocytes, endothelial cells and fibroblasts. Since it is difficult to reliably ...