Hücre ve Gelişim Biyolojisi çekmesi ekipman Bölümü kullanımı Marc Wozniak. Vanderbilt Üniversitesi Hücre Görüntüleme Paylaşımlı Kaynak. Bu çalışma DMB için ERP AHA hibe 12PRE10950005, DMB için AHA hibe 11GRNT7690040, NIH hibe R01 HL037675 tarafından desteklenmektedir. DMB Kardiyovasküler Araştırma Gladys P. Stahlman Başkanıdır.

Hayvanları kapsayan tüm işlemler uygun hayvan kullanımı ile onaylanmasını sağlayın ve vücut bakımı. 1. Advance Stok Tamponlar hazırlayın <ol><li> 2 L&#39;stok Ca2 + içermeyen Tyrode (Tablo 1) hazırlayın. NaOH ile 7.4 &#39;e pH ayarlayın. </li><li> 2 L&#39;stok ve 1.2 mM Ca2 + Tyrode (Tablo 2) hazırlayın. NaOH ile 7.4 &#39;e pH ayarlayın. </li></ol> Deney gününde 2. Perfüsvonun, Transferi ve Sindirim Tamponlar hazırlayın <ol><li> Ca 2 + Tyrode (Tablo 3), 150 ml perfüzyon tamponu hazırlayın ve bir 0.2 mikron filtre içinden süzme. </li><li> 35 ml hazırlayın Ca 2 + bedelsiz &quot;Tampon A&quot; &quot;Tampon B&quot; (Tablo 5) içeren (Tablo 4) ve 25 ml Ca 2 +. 0.2 mikron filtre aracılığıyla her filtre. </li><li> 0 çözümler hazırlayın.06, 0.24, 0.6, ve karıştırma Tampon A 1.2 mM Ca 2 + ve B Tablo 6 başına. </li><li> Fare vücut ağırlığı (BW) göre 25 mi perfüzyon tamponu (Tablo 7) olarak sindirim enzimleri eritilerek 25 mi sindirim tamponu hazırlayın. Kullanılmadan önce bir 0.2 mikron filtre içinden süzme. </li></ol> 3. Deney Düzeneği <ol><li> Mevcut piyasada bulunan ya da 3 ml&#39;lik bir akış oranı izin vermek için bir peristaltik pompa, boru ve bir ısı değiştirici sargı kullanılarak, sabit bir akış Langendorff cihazına monte / 37 ° C&#39;ye kadar ısıtıldı perfüzyon dak. Basit şematik Bell tarafından 2011 gözden sağlanan, ark 18 edilir. </li><li> Kanül dışarı akış 3 mL / dakikalık bir akış hızında 37 ° C üstünde olacak şekilde, Langendorff aparatındaki dolaşan su banyosu sıcaklığı (~ 47 ° C) ayarlayın. </li><li> Ultra saf tip I, 100 ml su, ardından 15 dakika boyunca perfüzyon sistemine,% 70 etanol ile çalıştırın. Sistemli Run perfüzyon tampon5 dakika boyunca m hava kabarcıklarını ortadan kaldırılmaktadır. </li><li> İstediğiniz bir yöntem ile tüm cerrahi aletleri sterilize. </li><li> Bir 23 G Luer adaptörü saplama ucu PE-50 boru kısa bir parça (yaklaşık 3 mm) takılarak kanül oluşturun. Boru ısı ucu aort yerleştirilmiş olacak, üzerinde bir dudak oluşturmak için. </li></ol> 4. kardiyomiyosit İzolasyon <ol><li> Intraperitoneal enjeksiyon ile, 0.2 ml heparin solüsyonu (1000 lU / ml) ile fare enjekte edilir. </li><li> 5 dakika sonra, izofluoran fare uyutmak. Tam (güçlü ayak tutam hiçbir yanıt) anestezi yaparken,% 70 etanol ile göğüs püskürtün. , Göğüs açın çabuk 4 ° C perfüzyon tamponunda aorta ve yer zarar vermemek için dikkatli olmak, kalbini kesip. Kavramak ve kalp altında kaldırmak için kavisli bir forseps kullanın. Bu ince makas ile kalbin arkasında ekleri kırpmaya alan oluşturur. </li><li> Yavaşça ince forseps iki çift aort tutup ve dikkatli bir şekilde openi çekerek kalp Cannulatekanülün dudak aort ng. Bu koroner damar yoluyla kalbin perfüzyon inhibe gibi, kanül ucu aort kapağı geçmiş ve ventrikül içine olmadığından emin olun. </li><li> Hemen kanülün dudak üzerinde aorta çevresinde 5-0 ipek sütür bir döngü bağlayarak kanüle aorta edin. </li><li> Not: 4.4 ile 4.2 mümkün olduğunca çabuk en kaliteli sindirim için tamamlanmalıdır Adımlar. </li><li> Not: Kalp daha önce perfüzyon tampon akan Langendorff cihazına takılı bir kanül üzerine direkt olarak monte edilebilir. Bununla birlikte, bir teşhir mikroskopu altında perfüzyon tamponu küçük bir şınngaya bağlanmış bir kanül üzerine kalp asılı çok başarılı olduğu bulunmuştur. Bu perfüzyon tamponu bir miktar koroner arterlerin temizlenmesi için takip ve güvenli bir şekilde kanül bağlı kalp sağlanması vasıtasıyla itilmesine izin verir. Sonra kanül perfüzyon tampon çalışan Langendorff&#39;a asılır. </li><li> Perfuse2-3 dakika boyunca 3 ml / dakikalık bir akış hızında serbest Ca2 + perfüzyon tamponu ile kalp. </li><li> Sindirim tampon enzim ve 7-10 dakika boyunca serpmek geçmek, kalp renkli yumuşak ve hafif olacak. </li><li> Not: Sindirim zaman enzim aktivitesi ve kalp büyüklüğüne göre ayarlanabilir. </li><li> Kalp palpably sarkık sonra, Tampon A (Ca 2 + bedelsiz tamponu) ile steril bir p60 çanak kanül ve yerden kalp kaldırmak. </li><li> Atriumlarm ve büyük damarları çıkarın. İsterseniz sağ ventrikül çıkarın. Forseps ile, küçük parçalar halinde ventrikül ayrı. 5 ml steril bir transfer pipet ile birkaç kez pipet başka hücreleri dağıtmak için. </li><li> 250 um naylon ağ filtre içerisinden 50 ml konik tüp içine hücre süspansiyonu filtre edin. </li><li> Süspansiyon, yaklaşık 10 dakika boyunca pelet izin verin. </li><li> 0.06 mM Ca 2 + çözeltisi içine pelet aktarın. Hücreler, 10 dakika boyunca pelet izin verin. En ölü hücreler yüzer olurken Sağlıklı hücreler, pelet olacaktır. </li&gt; <li> Süpernatanı havalandırın, ve 0.24 mM Ca 2 + çözeltisine pelet aktarın. </li><li> 10 dakika inkübasyon, aspirasyon tekrarlayın ve hücreler 1.2 mM çözelti haline gelene kadar kalan iki Ca 2 + çözümleri ile aktarın. Bu noktada, hücrelerin çoğunluğunun çubuk şekilli miyositler olmalıdır. </li></ol> 5. Ölçme Kasılma ve Kalsiyum Geçişler <ol><li> Not: kontraklil ve kalsiyum geçici Hem eş zamanlı olarak ölçülebilir. </li><li> 1 mg / ml arasında bir konsantrasyonda susuz DMSO içinde yeniden süspanse edilerek Fura-02:00 çözelti hazırlayın. Fura-02:00 solüsyonu, -20 ° C&#39;de bir desikatör içinde saklanabilir. </li><li> Yük 1 ul Fura-02:00 ile suspensiyon halindeki hücreler 1 ml. Hücreler koyu 10 dakika sonra aspire yüzer oturmak için izin verin. Hücreler yıkandı ve yıkamalar arasında 10 dakika boyunca karanlıkta pelet sağlayan, Ca 2 + Tyrode çözeltisi ile iki kez yıkayın. </li><li> Isıtmalı pe sağlayan bir sahne prospektüste bir cam lamelBir miyositlerdeki alan uyarıcı ile volta rfusion / aspirasyon ve elektrot. Ters mikroskop yerleştirin sahne insert bir immersiyon yağı hedefi kullanıyorsanız lens petrol bir damla ekleyerek, floresan ölçümü için ayarlanmış. </li><li> 1.2 mM Ca 2 + tirod çözeltisi ısıtmalı yerçekimi perfüzyon kurmak sahnede çözelti 37 ° C ulaşır böylece. Vakum aspirasyon takın. Elektrotlar takın. </li><li> Mikroskop sistemi açın ve rasyometrik floresan ve hücre boyutlandırma veri toplama yazılımı açın. </li><li> An perfüzyon engelleme sağlıklı hücreleri yerleşmek için izin, sahne inserte Fura-02:00 yüklü hücrelerin birkaç damla ekleyin. </li><li> Istenen hücreyi bulmak için 40X objektif kullanın. Sağlıklı bir hücre çubuk şeklinde ve kendiliğinden sözleşme olmamalıdır. Bu gözle görülür ve eşit sözleşme cep 5-20 V&#39;de tempolu gerektiği zaman </li><li> Not: Fizyolojik analizi, yüksek kaliteli parçalanıp sindirilmesi gerekmektedir. Bu sağlıklı görünüyorsun ama pa yok hücrelere sahip olmaları mümkündür<html> fizyolojik analizi için de ce. Sindirim protokol, yüksek kaliteli, sağlıklı miyositleri almak için arıtma gerekebilir. Ayrıca, mutant veya hastalıklı kalpleri protokol değişiklikleri gerektirebilir. </li><li> Objektif rotasyon ve değişim diyafram böylece istenilen hücre ekran ve arka plan üzerinde yatay dizilmiş olan Ayarı görünür değil. </li><li> Miyositinin her sonuna kadar kenar algılama barlar hizalayın ve eşiği ayarlayın. Düzgün sarkomer ile hücrenin bir kısmı üzerinde sarkomer algılama hizalayın. Artık size bu bar yapabilirsiniz, sarkomer uzunluğunun daha doğru matematiksel model bu kadar uzun sarkomer iki popülasyonları üzerinde değil gibi olacaktır. </li><li> Pace hücreleri 2 Hz ve kayıt önce 10-15 saniye boyunca 5-20 V. </li><li> Kayıt izleme. Ortam ışığı iyi kalsiyum geçici kayıt için minimize edilmelidir. </li></ol> 6. Analiz <ol><li> Rasyometrik floresan ve hücre boyutlandırma veri toplama yazılımı kullanarak izlerini analiz edin. </li></ol>title &quot;&gt; 7. Diğer Çalışmalar <ol><li> Fiksasyon ve immün, doğrudan ilaç etkileri, kısa vadeli kültürü ve atomik kuvvet mikroskobu değerlendirirken: Bu protokol dahil olmak üzere diğer deneyler çeşitli için kullanılabilir fazla hücreleri sağlar. </li></ol>

<ol>
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D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FGF23+is+a+novel+regulator+of+intracellular+calcium+and+cardiac+contractility+in+addition+to+cardiac+hypertophy.">FGF23 is a novel regulator of intracellular calcium and cardiac contractility in addition to cardiac hypertophy.</a> American Journal of Physiology - Endocrinology and Metabolism. , (2013).</li><li>Helmes, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Titin+Determines+the+Frank-Starling+Relation+in+Early+Diastole.">Titin Determines the Frank-Starling Relation in Early Diastole.</a> The Journal of General Physiology. 121, 97-110 (2003).</li><li>Bell, R. M., Mocanu, M. M., Yellon, D. 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Biz ifşa hiçbir şey yok.

Kalite kardiyomiyositlerin izolasyonu uygulama ve optimizasyon bazı düzeyi gerektirir. Bu protokol büyük ölçüde sindirim sonucunu etkileyecek önemli adımları içermektedir. Performans ve protokol giderirken bu dikkatle düşünülmelidir. Kanülasyon ve perfüzyon için kalp kaldırılması süre az, ideal olarak, beş dakikadan daha aza düşürülmesi gerekmektedir. Buz soğukluğunda tamponda kalp fare çıkarılması ve kesme ve kanüle sonra buz soğukluğunda perfüzyon tampon maddesi içinde hemen kalp ...

Cardiomyocytes provide the contractile force for the heart. Each myocyte contains organized cytoskeletal and contractile elements essential to contraction. The heart not only contains cardiomyocytes but also fibroblasts, connective tissue, and modified myocytes such as Purkinje fibers. Changes in any or all of these cell types can be seen in heart failure, making determination of the ultimate cause of heart dysfunction difficult. Decreased ventricular contractility of a failing heart is preceded by varying degrees of fibrous tissue build-up and hypertrophy and dysfunction of the cardiomyocytes1. Arrhythmias can be caused by dysfunction in the conducting cells2, fibrosis interrupting cardiac conduction3, or changes in the expression of ion channels in cardiomyocytes1,4. Indeed, these problems often co-exist in a complicated fashion.
Isolating and studying cardiomyocytes allows one to examine contractile and electrical dysfunction of individual myocytes in the context of overall heart function. Techniques to isolate individual cardiac myocytes were developed over 40 years ago in an effort to better study physiology of this &#8216;workhorse&#8217; cell of the heart. Isolation techniques have been improved to increase yield and quality of cells, with the vital development of coronary perfusion via cannulation of the aorta, first performed in 19705. This article describes cardiomyocyte isolation followed by measurement of contractility and calcium transients using ratiometric fluorescence and cell dimensioning data acquisition software. In both genetic and environmental models of heart disease, utilizing this protocol provides key information about cell contractility, sarcomeric contractility and relaxation, calcium transients, and cytoskeletal disruption. This system is often used to test the effect of drugs on isolated cardiomyocytes6,7. We employ the system to compare contractility and calcium transients of cardiomyocytes between wild type mice and mice with a mutation leading to dilated cardiomyopathy.
Studying individual cardiac myocytes has numerous advantages over other commonly used methods to study cardiac anatomy and physiology. Echocardiography and electrocardiography provide information about the contractile function and electrical conduction of the heart overall. Neither of these methods explains the cause or nature of the dysfunction below the level of the whole organ. Measuring contractility of individual cardiomyocytes using edge-detection and sarcomere length algorithms can demonstrate that changes within the contractile cell of the heart are present within global heart dysfunction. Measuring calcium transients demonstrates the influence of changes in ion flux at a cellular level to contractile dysfunction or arrhythmias.
In addition to complementing whole organ studies, this protocol provides cells easily stained for sub-cellular components. It is possible to see cytoskeletal changes by staining tissue sections of the whole heart; however, one is only able to visualize a cross section of cells. A whole cardiomyocyte is thicker than the typical tissue section, and given the arrangement and length of the cardiomyocytes, it is difficult to see whole cells in a single section. Isolated cardiomyocytes can be fixed immediately and stained for a variety of cytoskeletal elements and ion channels. Confocal images can be compiled into full thickness z-stacks of the cell. Additionally, staining isolated cardiac myocytes allows for staining of components that are impossible to visualize in sections, such as the membrane invaginations known as t-tubules.
Cultured cell lines of cardiac myocytes do exist, some of which have similar transcriptional profiles and phenotypic features to cardiac myocytes. Some, such as the HL-1 cell line, even retain some sarcomeric organization and rudimentary contractile ability8. Despite these qualities, cultured cells lack the &#8220;box car&#8221; shape and t-tubule structure of cardiomyocytes and have less sarcomeric organization. These components are essential to the function of cardiomyocytes in vivo. In addition, using cultured cells allows genetic manipulation but not in vivo physiologic manipulation. Isolated cardiomyocytes retain their cellular organization long enough to study but are still easily imaged and even transduced9. 
Isolated cardiomyocytes are extremely versatile, providing a substrate to identically analyze cells from varied experimental conditions. Isolation and physiological analysis of myocytes have been used by countless labs for a variety of experiments including studying the effect of drugs6,7 or small molecules10, environmental stressors11, infection12, illness13, or genetic mutation14,15 on contractility and/or calcium transients and studying the regulation of contractility and calcium release within a cardiomyocyte15-17. With the application of any of these stressors, changes in heart function could be due to any combination of changes in the cardiomyocytes themselves and changes in the surrounding environment of the heart, such as scarring, changes in conductivity, or changes to the extracellular matrix. This is an ideal protocol for answering research questions regarding individual myocyte structure and function in the context of heart failure from any cause.
Although it has countless advantages over other techniques, physiological analysis of isolated cardiomyocytes is not ideal for every research question. Isolation is a terminal procedure, thus cardiomyocytes can only be assessed at one time in the mouse&#8217;s life. This can be partially overcome using cohorts and sacrificing individual mice along the course of illness development or by monitoring the mouse to determine whether it has clinically significant heart failure before isolation. Although this protocol can determine whether cardiomyocyte dysfunction has occurred, one cannot draw the conclusion that this dysfunction is the root cause of heart failure without additional experimental information. It is possible that cardiomyocyte dysfunction itself is secondary to other changes in the heart. Despite these challenges, studying cardiomyocytes offers valuable information in determining the nature of dysfunction in the heart, especially when combined with other experiments. 
The following protocol is adapted from one provided by Dr. Chee Lim, Vanderbilt University. Although contractility and calcium flux data are highly reproducible and useful in comparing hearts with genetic or environmental manipulation, isolation itself remains a technique dependent protocol, requiring optimization. Minimizing the time between removing the heart from the mouse and perfusing the heart via aortic cannulation is essential for a quality digestion. In addition, digestion time and enzyme concentration can be optimized for the highest quality myocytes. Steps likely requiring optimization, along with suggestions for optimization, are noted below.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">NaCl</td>
			<td style="width:221px;">RPI</td>
			<td style="width:159px;">S23020</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">KCl</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">P-3911</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:253px;">MgCl2</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">M-8266</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">HEPES</td>
			<td style="width:221px;">EM Science</td>
			<td style="width:159px;">S320</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:253px;">NaH2PO4</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">S5011</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:253px;">CaCl2 &#376; 2H2O</td>
			<td style="width:221px;">Fisher</td>
			<td style="width:159px;">C79</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">D-(+)-Glucose</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">G7528</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">2,3-Butanedione-monoxime (BDM)</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">B-0753</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Taurine</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">T-0625</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">A-7030</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Collagenase B</td>
			<td style="width:221px;">Roche Diagnostics</td>
			<td style="width:159px;">1-088-823</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Collagenase D</td>
			<td style="width:221px;">Roche Diagnostics</td>
			<td style="width:159px;">1-088-882</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Protease XIV</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">P-5147</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Heparin</td>
			<td style="width:221px;">APP Pharmaceuticals, LLC</td>
			<td style="width:159px;">401586D</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Isofluorane</td>
			<td style="width:221px;">Butler Schein</td>
			<td style="width:159px;">11695-6776-2</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">Fura-2 AM</td>
			<td style="width:221px;">Teflabs</td>
			<td style="width:159px;">103</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">DMSO &#8211; anhydrous</td>
			<td style="width:221px;">Sigma</td>
			<td style="width:159px;">276855</td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:253px;">IonOptix cell pacing and fluorescent analysis system</td>
			<td style="width:221px;">IonOptix</td>
			<td style="width:159px;"></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">250 um nylon mesh filter</td>
			<td style="width:221px;">Sefar America</td>
			<td style="width:159px;">Lab Pak 03-250/50</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">23G leur-stub adaptor</td>
			<td style="width:221px;">Becton Dickinson</td>
			<td style="width:159px;">1482619E</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:253px;">PE-50 tubing</td>
			<td style="width:221px;">Becton Dickinson</td>
			<td style="width:159px;">427410</td>
			<td></td>
		</tr>
	</tbody>
</table>

isolation and physiological analysis of mouse cardiomyocytes

Cardiomyocytes, the workhorse cell of the heart, contain exquisitely organized cytoskeletal and contractile elements that generate the contractile force used to pump blood. Individual cardiomyocytes were first isolated over 40 years ago in order to better study the physiology and structure of heart muscle. Techniques have rapidly improved to include enzymatic digestion via coronary perfusion. More recently, analyzing the contractility and calcium flux of isolated myocytes has provided a vital tool in the cellular and sub-cellular analysis of heart failure. Echocardiography and EKGs provide information about the heart at an organ level only. Cardiomyocyte cell culture systems exist, but cells lack physiologically essential structures such as organized sarcomeres and t-tubules required for myocyte function within the heart. In the protocol presented here, cardiomyocytes are isolated via Langendorff perfusion. The heart is removed from the mouse, mounted via the aorta to a cannula, perfused with digestion enzymes, and cells are introduced to increasing calcium concentrations. Edge and sarcomere detection software is used to analyze contractility, and a calcium binding fluorescent dye is used to visualize calcium transients of electrically paced cardiomyocytes; increasing understanding of the role cellular changes play in heart dysfunction. Traditionally used to test drug effects on cardiomyocytes, we employ this system to compare myocytes from WT mice and mice with a mutation that causes dilated cardiomyopathy. This protocol is unique in its comparison of live cells from mice with known heart function and known genetics. Many experimental conditions are reliably compared, including genetic or environmental manipulation, infection, drug treatment, and more. Beyond physiologic data, isolated cardiomyocytes are easily fixed and stained for cytoskeletal elements. Isolating cardiomyocytes via perfusion is an extremely versatile method, useful in studying cellular changes that accompany or lead to heart failure in a variety of experimental conditions.

Kontraklil ve geçici kopyalamaları (Şekil 2) toplandığında, verileri kolayca uygun yazılım ile analiz edilir. Bütün izleme, veya bunların kısımları (Şekil 3) ortalama edilebilir. Kasılma çeşitli yollarla analiz edilebilir. Sistolik fonksiyonu için, bir kısalma zirve fraksiyonel kısalma, ya da zamanla daralma hızı ile kasılma büyüklüğünü ve daralma hızı değerlendirmek. Diyastolik fonksiyon% 50 kısalma ve gevşeme hızı zaman ile benzer şekilde analiz e...

Watch this Scientific Journal Video about Isolation and Physiological Analysis of Mouse Cardiomyocytes at JoVE.com

Isolation and Physiological Analysis of Mouse Cardiomyocytes

Individual cardiomyocytes from wild type and mutant mice can be isolated from the heart in order to study their contractility and calcium transients. This allows characterization of the contribution of cellular dysfunction to heart dysfunction from any cause.

Fare kardiyomiyositlerde İzolasyon ve Fizyolojik Analizi

Cardiomyocytes, the workhorse cell of the heart, contain exquisitely organized cytoskeletal and contractile elements that generate the contractile force ...

isolation-and-physiological-analysis-of-mouse-cardiomyocytes

Research

JoVE Journal

Biology

22.8K Görüntüleme.  Vanderbilt University.  Individual cardiomyocytes from wild type and mutant mice can be isolated from the heart in order to study their contractility and calcium transients. This allows characterization of the contribution of cellular dysfunction to heart dysfunction from any cause. 