MAK kabul eder bu işten Ulusal Sağlık Enstitüleri için destek / Ulusal Kanser Enstitüsü vermek sayı - 1R15CA152985-01A1. Bu proje aynı zamanda Miami Üniversitesi lisans Araştırma Ödülü programı, Miami Üniversitesi doktora-lisans fırsatları burs programı ve Miami Üniversitesi yaz bilim programı tarafından desteklenmiştir.

videoda yürütülen ve aşağıda açıklanan yordamları kurumsal hayvan bakım ve kullanım Komitesi (IACUC) Miami Üniversitesi'nde tarafından onaylanmış olan.
 1. hazırlanması ve uyarıcı Test <ol><li>Ağımdaki iki farklı alanlara cerrahi işlem, ameliyat masasına ve sonrası operasyon Masası için. Her iki alanda tüm malzeme ve gerekli aletleri ile sahne.
	Bacalı bir başlık altında <ol><li>sahne işletim tablo. Yordamı sürekli ve engelsiz performansını sağlayan bir şekilde ekipman ile tablo düzenlemek.</li>
		<li>Ameliyat sonrası bir tablo aynı odada ve operasyonun ana tablo yakınındaki kurmak. Prosedür boyunca steril ortam olarak her iki tabloları korumak.</li>
	</ol></li> <li>Yer şu ameliyat masasında sağlar: bir cam kavanoz kapağı, bir 15 mL tüp, Cerrahi makas, % 70 etanol, iki köpük panoları, iki forseps, iki 1 mL 21 ölçer şırınga, iki 50 mL tüp, bir santrifüj tüpü bir sıkma şişe bir çift ile , bir kriyojenik şişe, dört cerrahi yastıkları, on iğne, sterilize mendil dispenser ve "Sharps" konteyner.</li>
	<li>Sonrası işlemi masaya aşağıdaki malzemeleri koyun: bir analitik denge, bir 4 L dewar sıvı azot, sığ geniş ağız dewar, kayan bir microtube raf, bir cerrahi makas çifti, iki forseps, dört kriyojenik şişeleri ve bir dispenser steril mendil.</li>
	<li>Doldurmak bir 50 mL ve bir 15 mL tüp % 75 birime formalin ile.</li>
	<li>Her köşede bir PIN kullanarak bir cerrahi pedi diseksiyon panosu olarak hizmet etmek bir yaklaşık 30 cm x 30 cm köpük kuruluna yapıştırmayın. Kalan dört pimleri işlem sırasında kullanın. Organları ameliyat sonrası tabloya aktarmak için cerrahi bir yastık ile daha küçük bir köpük tahta hazırlayın. 
	Dikkat: Isoflurane (% 99.9) bir toksik kimyasal ve havalandırma mahallede Emanet atık anestezik gaz 8 atma gelen en yüksek düzeyde sağlamak için kullanılmalıdır. Araştırmacılar isoflurane kullanarak açık bırak yöntemi ile ilişkili riskleri hakkında daha fazla bilgi Taylor ve Mook 8 tarafından bir makalede bulunabilir.</li>
	<li>Cam kavanoz ve emmek isoflurane (% 99.9) birkaç damla ile bir cerrahi pedi yerleştirin ve fare isoflurane ile doğrudan temas önlemek için üst üzerinde bir kağıt havlu yer. Benzer şekilde, kalan boru hattı ve isoflurane birkaç damla ile ıslatın ve ek bir pad fare ve isoflurane arasında doğrudan temas önlemek için üst üzerinde yer için bir cerrahi pedi kullanın.</li>
	<li>Maksimum doldurana kadar hattı dewar ulaşıldığında sığ geniş ağız içine dökün sıvı nitrojen.</li>
	<li>Yer diseksiyon anestezi içine seçilmiş fare odası, Cam Kavanoz ile beraber batırılmış bir yastık bırakır ~ 1 dakika süreyle kapak ile örtülü isoflurane (% 99.9), Yani 
	Not: Bu süre fare fare değişir. Fare bilinçsiz olduğunda, odasından çıkarın ve çalışma kartı üzerine yerleştirin.</li>
	Ventral tarafı yukarı ve onun baş bilim adamı uzak işaret ile yalan söylüyor <li>fare gelecek şekilde yönlendirin. Kafasının isoflurane (% 99.9) birkaç damla ile batırılmış bir cerrahi pedi ile kaplı tüp içine yerleştirin ve fare uyaranlara yanıt vermeyen olduğundan emin olmak için bir ayak çimdik tarafından bir uyarıcı testi gerçekleştirin.
	<ol><li>Bu sınama başarısız olursa ve fare ayak çimdik testine yanıt verir, adımları yineleyin 1.8.</li>
	</ol></li></ol> 2. İlk kesi, kalp ponksiyon ve ötenazi <ol><li>cerrahi köpük kartına fare ekstremitelerin Pin ve fare ile % 70 etanol ventral tarafına ıslak.</li>
	<li>Kürk/cilt üretral açılış yakınındaki Forseps ile pinch ve biraz yukarı doğru çekin. Bir kesi ile karın boşluğu üretral açılması orta hat üzerinden başlayıp çene Cerrahi makas ile kesme yapmak.
	<ol><li>İlk belgili tanımlık kesme başlangıç noktasının yakınını, kürk/deri bir taraftan Forseps ile tut ve başka bir kesi makas ile aşağı ve çapraz olarak doğru arka pençe yapmak.</li>
		<li>Bu şekilde karşı tarafta tekrarlayın. 
		Not: Kürk/cilt daha geniş bir açıklık oluşturmanızı sıkıştırdık, ancak gerekli değildir.</li>
	</ol></li> <li>Kalp bulun ve kesesi çevresinde şırınga iğne tıkanmasını önlemek için kalp, kalp zarını kaldırın.
	<ol><li>Kalp zarını Forseps ile kavramak ve makasla kesti. Kalp ponksiyon dikkatli kalbi şırınga iğne ekleyerek gerçekleştirmek ve pistonu geri çekmek yavaş yavaş başlamak.</li>
		<li>Kalbin pompalama eylemi taklit ve çok hızlı çizim önlemek için iğne pistonu en uygun kan toplanması için kullanılmasını. 
		Not: Genellikle 1 mL kan toplanabilir.</li>
		<li>Kan toplama işlemini tamamladıktan sonra kan santrifüj tüpü ve "Sharps" konteyner içine şırınga atmayın içine ortadan kaldırmak.</li>
		Kalbi delik gerçekleştirildikten sonra <li>kalp bağlanma ekleri kaldırarak ötenazi taşımak. 
		Not: Bu yordam belirli bu çalışmada serum koleksiyonu için koagüle kan izin vermek için kalbi delik önce şırıngada Heparin, bir anti-koagulant eklenmedi. Araştırmacı plazma toplamak için kan pıhtılaşması önlemek isterseniz, ancak, heparin kalbi delik önce şırınga için eklenebilir.</li>
	</ol></li> <li>Çalışma Genotipleme fare gerektirir, bir bölümü Kulak makasla makasla kesme ve yer içine bir santrifüj tüpü genotip doğrulama için.</li></ol>
 3. pankreas çıkarma <ol><li>bulun mide fare sol tarafında. Begin yavaşça (yırtılma önlemek için) pankreas mide ve oniki parmak bağırsağı ayıran iki forseps kullanarak. 
	Not: mide ve bağırsak pankreas ayırma, o forseps pankreas dokusu organları uzak yol değil ezmek, veya pankreas Forseps ile gözyaşı hafifçe kullanılır çok önemlidir.
	<ol><li>Pankreas ince bağırsak değişir ve ileum bölümlerine ayırmak devam etmek ve son olarak gelen kalın bağırsak caecum.</li>
	</ol></li> <li>Caecum, forseps yeniden konumlandırmak ve pankreas boyunca kalan kolon rektum doğru ayrılması devam. 
	Not: Bu noktada, bu kesmek ve kolon rektum hemen önceki bölgesine mide bölümü kaldırmak uygundur.</li>
	<li>Pankreas bulun ve dalak bağlı. Pankreas fareyi sağ tarafına doğru kaydırın. Pankreas ve göğüs boşluğuna arasında kalan bağlantı Forseps ile tam olarak pankreas ve dalak keşfedemiyoruz ayırmak için ayırmak.</li>
	<li>Pankreas kaldırmak ve sınav için dışarı yayılır. Tanıtım amaçlı pankreas bağlı dalak bırakın.
	<ol><li>Tümünü Kaldır bağ dokusu, pankreas şişman ve Mezenterik dokudan. 
		Not: Bu doku daha beyaz renkte ve böylece kolayca ayırt edici renkte pembe pankreas dokusundan olabilir. Bütün pankreas kaldırılması gerekiyor, bu özellikle önemlidir. Örneğin, pankreas tartılır ve vücut ağırlığı için ya da hayvan grupları arasında karşılaştırıldığında gerekiyor. Ptf1a içinde cre / +; LSI-Kras G12D / + fare modeli, özellikle içinde belgili tanımlık büyük ay, zor fibröz pankreas dokusu mevcut olabilir. Bu durumda, bağırsak tümör dokusunda titreşimli gibi dikkatli pankreas kaldırılması yürütülmelidir. İleri durumlarda, anormal dalak ve karaciğer dokusu de gözlenebilir.</li>
	</ol></li> <li>İsterseniz, diğer organlara bu noktada kaldırmak.</li></ol>
 4. veri toplama ve depolama <ol><li>sonra ayıklama organların Ģunları alanının korunmasına ilişkin örnekler taşımak.</li>
	<li>Her organ tartmak ve onların anılan sıraya göre kriyojenik şişe koyun. 
	Not: zekâh kitle her örnek, daha sonra başvurmak için herhangi bir usulsüzlük kaydedilmelidir.</li>
	<li>Bir kez organları tartılır vardır, ek dondurma için sıvı azot içine koyup.</li>
	<li>Buz gibi ek sonra-80 ° C'de organları uzun süreli depolama için mağaza.</li>
	<li>Yer depolanan formalin örnekleri tezgah üstüne gece ve ertesi sabah değiştirmek onların çözüm formalin % 70 etanol için. 
	Not: Bu örnekler uzun süreli depolama için 4 ° C'de muhafaza edilmelidir.</li>
	<li>Uzun süreli depolama için kan ve kulak yumruk-80 ° C'de dondurmak. Kan serumu koleksiyonundan için kan koyulaştırmak için 30 dk sonra bu santrifüj kapasitesi izin. Bir pipet kullanarak serum bölümü kaldırmak ve sonra-80 ° C.</li>
</ol> 5. Clean Up <ol><li>Sanitize tüm diseksiyon araçları sterilizing mendil ile. Cap isoflurane ile kaplı tüp cerrahi pedi batırılmış. Köpük gemide cerrahi pedi taze bir cerrahi pedi ile değiştirin. Tesisin toplanan değil fare bölümlerini elden &#39; un hayvan elden çıkarma ilkesinin.</li>
</ol>

<ol>
	<li>Fesinmeyer, M. D., Austin, M. A., Li, C. I., De Roos, A. J., Bowen, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+Survival+by+Histologic+Type+of+Pancreatic+Cancer.">Differences in Survival by Histologic Type of Pancreatic Cancer.</a> Cancer Epidemiol Biomarkers Prev. 14 (7), 1766-1773 (2005).</li><li>Jackson-Grusby, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+cancer+in+mice.">Modeling cancer in mice.</a> Oncogene. 21 (35), 5504-5514 (2002).</li><li>Heid, I., 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=Early+requirement+of+Rac1+in+a+mouse+model+of+pancreatic+cancer.">Early requirement of Rac1 in a mouse model of pancreatic cancer.</a> Gastroenterology. 141 (2), 719-730 (2011).</li><li>Hingorani, S. R., 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=Preinvasive+and+invasive+ductal+pancreatic+cancer+and+its+early+detection+in+the+mouse.">Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse.</a> Cancer cell. 4 (6), 437-450 (2003).</li><li>Shi, C., 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=KRAS2+Mutations+in+Human+Pancreatic+Acinar-Ductal+Metaplastic+Lesions+Are+Limited+to+Those+with+PanIN+Implications+for+the+Human+Pancreatic+Cancer+Cell+of+Origin.">KRAS2 Mutations in Human Pancreatic Acinar-Ductal Metaplastic Lesions Are Limited to Those with PanIN Implications for the Human Pancreatic Cancer Cell of Origin.</a> Mol Cancer Res. 7 (2), 230-236 (2009).</li><li>LaConti, J. J., 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=Distinct+serum+metabolomics+profiles+associated+with+malignant+progression+in+the+KrasG12D+mouse+model+of+pancreatic+ductal+adenocarcinoma.">Distinct serum metabolomics profiles associated with malignant progression in the KrasG12D mouse model of pancreatic ductal adenocarcinoma.</a> BMC Genomics. 16 (1), 1-10 (2015).</li><li>Ludwig, M. R., 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=Surveying+the+serologic+proteome+in+a+tissue-specific+kras(G12D)+knockin+mouse+model+of+pancreatic+cancer.">Surveying the serologic proteome in a tissue-specific kras(G12D) knockin mouse model of pancreatic cancer.</a> Proteomics. 16 (3), 516-531 (2016).</li><li>Taylor, D. K., Mook, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+Waste+Anesthetic+Gas+Concentrations+Associated+with+the+Open-Drop+Method.">Isoflurane Waste Anesthetic Gas Concentrations Associated with the Open-Drop Method.</a> J Am Assoc Lab Anim Sci. 48 (1), 61-64 (2009).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Pharmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Li, D. S., Yuan, Y. H., Tu, H. J., Liang, Q. L., Dai, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+protocol+for+islet+isolation+from+mouse+pancreas.">A protocol for islet isolation from mouse pancreas.</a> Nature Protocols. 4 (11), 1649-1652 (2009).</li><li>Auer, H., 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=The+effects+of+frozen+tissue+storage+conditions+on+the+integrity+of+RNA+and+protein.">The effects of frozen tissue storage conditions on the integrity of RNA and protein.</a> Biotech Histochem. 89 (7), 518-528 (2014).</li><li>Ma, J., Leung, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limbic+System+Participates+in+Mediating+the+Effects+of+General+Anesthetics.">Limbic System Participates in Mediating the Effects of General Anesthetics.</a> Neuropsychopharmacology. 31, 1177-1192 (2006).</li><li>Ijichi, H., 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=Aggressive+pancreatic+ductal+adenocarcinoma+in+mice+caused+by+pancreas-specific+blockade+of+transforming+growth+factor-&#946;+signaling+in+cooperation+with+active+Kras+expression.">Aggressive pancreatic ductal adenocarcinoma in mice caused by pancreas-specific blockade of transforming growth factor-&#946; signaling in cooperation with active Kras expression.</a> Genes Dev. 20 (22), 3147-3160 (2006).</li><li>Abiatari, I., 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=Moesin-dependent+cytoskeleton+remodelling+is+associated+with+an+anaplastic+phenotype+of+pancreatic+cancer.">Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.</a> J Cell Mol Med. 14 (5), 1166-1179 (2010).</li><li>Goodpaster, A. M., Romick-Rosendale, L. E., Kennedy, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=a+Statistical+significance+analysis+of+nuclear+magnetic+resonance-based+metabonomics+data.">a Statistical significance analysis of nuclear magnetic resonance-based metabonomics data.</a> Analytical biochemistry. 401, 134-143 (2010).</li><li>Yadav, 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=Idiopathic+Tumefactive+Chronic+Pancreatitis:+Clinical+Profile,+Histology,+and+Natural+History+After+Resection.">Idiopathic Tumefactive Chronic Pancreatitis: Clinical Profile, Histology, and Natural History After Resection.</a> Clin Gastroenterol Hepatol. 1 (2), 129-135 (2003).</li></ol>

Yazarlar ifşa gerek yok.

Mevcut yöntemler açısından önemi Fare diseksiyonlarının resmi olmayan diğer videolar var iken, ilk profesyonel kalitede video bu makale sağlar, akran gözden, görsel gösteri tüm çıkış için gerekli ve fare pankreas diseksiyon10 tarafından hasat ayrıntılı adımlar . Metabolik aktivite ve insülin için ana bir organ olan pankreas ile üretim, diseksiyon ve pankreas hasat sağlayan fizyolojik özellikleri11korunması için. Pankreas izo...

Fare insan pankreas kanseri1,2önemli bir hayvan modeli olarak ortaya çıkmıştır. Ptf1a içindecre / +; LSI-KrasG12D / + fare modeli, sıçan sarkomu (K-Ras) onkogen pankreas İntraepitelyal Neoplazi (PanINs), bu ilerleme bilinen pankreasın prekanseröz lezyonlar inisiyasyon sonuçlanan pankreasın sadece aktif Kristen pankreas duktal Adenokanserine için genellikle PDACs3adlandırılır. Bu fare modeli sistemi insan pankreas kanseri4,5, PanINs hayat ve sık sık PDAC içinde ilerleme ilk beş ay içinde ortaya ek avantajı için en iyi mevcut hayvan modellerinden birini sağlayan bir Pankreas kanseri en sık 60-70 yaşın insanlarda ortaya çıkar, ancak yıl4,5, tek.
Pankreas diseksiyon Ptf1a dan tarafından çıkarılmasıcre / +; LSI-KrasG12D / + fareler çeşitli yaş itibariyle sağlar detaylı boyuna histolojik muayene pankreas kanseri gelişimi için en erken PanIN aşamaları ilerleme PDAC3,4 ' e değişen , 5. pankreas yaşları 5 ila 15 ay arasında değişen, hasat da kullanılabilir doku özleri pankreas4 metabolizması sağlıklı hastalıklı doku geçiş sırasında oluşan küresel değişiklikleri tanımlamak için hazırlamak için 6,7.
Bu yazı bir fare pankreas çekimi gerçekleştirmek için gereken adımları, tam bir görsel kılavuzu sunar ve daha fazla çözümleme için bir pankreas depolamak için yönergeler sağlar. Bu kılavuz-ecek var olmak aynı derecede değerli bireyler de dahil olmak üzere diğer pankreas Hastalıkları Araştırma tip ı diyabet ve özellikle öğrenciler ve araştırmacılar fare pankreas kullanarak hasat içeren araştırma için yeni yararlı olmalıdır diseksiyon metabolik profil oluşturma veya histolojik analizler için.

<table border="0" cellpadding="0" cellspacing="0" width="790">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass Jar&#160;</td>
			<td style="width:288px;">Corning</td>
			<td style="width:119px;">3140-150</td>
			<td style="width:167px;">The glass jar used in the video has been discontinued.&#160; This is its replacement.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Lid of Glass Jar</td>
			<td style="width:288px;">Corning</td>
			<td style="width:119px;">9985-150</td>
			<td>The glass jar lid used in the video has been discontinued.&#160; This is its replacement.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">15 mL Falcon Tubes</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">339650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical Scissors</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">9201</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Squeeze bottle&#160;</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">03-409-10DD</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">100% Ethanol</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">22-032-103</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Formalin</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">245-684</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Foam Boards</td>
			<td style="width:288px;">Therapak</td>
			<td style="width:119px;">562908</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Forceps</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">200205SHN</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1 mL 21G Syringes</td>
			<td style="width:288px;">BD Biosciences</td>
			<td style="width:119px;">309624</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50 mL Falcon Tubes</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">339652</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">2.0 mL Microcentrifuge Tubes&#160;</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">02-681-258</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgical Pads</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">S67011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">T-Pins Length:&#160; 2&#34;&#160;</td>
			<td style="width:288px;">Advance Store Products</td>
			<td>X32T-05</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sterilizing Wipes</td>
			<td style="width:288px;">Professional Disposables International Inc.</td>
			<td style="width:119px;">Q85084</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Sharps Container</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">14-827-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Analytical Balance</td>
			<td style="width:288px;">Marshall Scientific</td>
			<td style="width:119px;">ME-AE200</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">4L Dewar</td>
			<td style="width:288px;">Taylor-Wharton</td>
			<td style="width:119px;">4LD</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Shallow Wide Mouth Dewar</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">F3087-V</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Floating Microtube Rack</td>
			<td style="width:288px;">VWR</td>
			<td style="width:119px;">60986-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Cryogenic Vial 1.2 mL, Sterile</td>
			<td style="width:288px;">Fisher Scientific</td>
			<td style="width:119px;">10-500-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Isothesia (Isoflurane)&#160;</td>
			<td style="width:288px;">Henry Schein Animal Health</td>
			<td style="width:119px;">050033</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Liquid Nitrogen</td>
			<td style="width:288px;">Wright Brothers</td>
			<td style="width:119px;">NIT-60-XX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mouse Kras Strain</td>
			<td style="width:288px;">The Jackson Laboratory</td>
			<td style="width:119px;">OO8179</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Mouse Cre Strain</td>
			<td style="width:288px;">MMRRC</td>
			<td style="width:119px;">OOO435-UNC</td>
			<td></td>
		</tr>
	</tbody>
</table>

dissection of the mouse pancreas for histological analysis and metabolic profiling

Biz pankreas belirli transkripsiyon faktörü, 1a cre-recombinase soruşturma edilmiştir; füme balığı-dur-lox-Kristen sıçan sarkomu, 12 kodon aspartik asit glisin (Ptf1acre / +; LSL-KrasG12D / +) fare zorlanma insan pankreas kanseri bir model olarak. Bizim mevcut çalışmalar amacı pankreas kanseri ilerleme roman metabolik biyolojik belirlemektir. İdrar, dışkı, kan ve pankreas dokusu özler yanı sıra pankreas kanseri ilerleme yerleştirmeye histolojik analizler metabolik profil oluşturma gerçekleştirdiniz. Fare pankreas insanlar gibi iyi tanımlanmış bir solid organ değil, ama oldukça kolayca bireyler ile fare iç anatomi yabancı veya az veya hiç deneyimi gerçekleştirme sahip bireyler tarafından tanımlanmış değildir havza dağıtılmış bir yumuşak doku fare organ diseksiyonu. Bu makalenin amacı ayrıntılı bir step-wise sağlamaktır diseksiyon tarafından fare pankreas kaldırma yeni başlayanlar Kılavuzu için görsel gösteri. Bu madde öğrenciler ve araştırmacılar yeni özellikle değerli diseksiyon metabolik profil oluşturma veya histolojik analizler için tarafından fare pankreas hasat gerektirir araştırma olmalıdır.

Şekil 1 çalışma çevre bölgesi özetini gösterir ve Şekil 2 post işlem alanı gösterir. Süre bu ayar donanımları en az miktarda sağlar ve evreleme, bireylerin bu bireysel gereksinimlerinizi en iyi karşılayacak şekilde değiştirmek tercih edebilir. Protokol deneme belirli ihtiyaçlarına göre optimize edilmelidir. Bu yordamı uygun euthanization9gerektiren bir fare hayat sonlandıran bir ?...

Watch this Scientific Journal Video about Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling at JoVE.com

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

Video bu makalede ayrıntılı bir gösteri diseksiyon histolojik analizi ve metabolik profil oluşturma için başarıyla bir fareden pankreas kaldırmak için gerekli yordamlar sağlar.

Fare pankreas diseksiyon histolojik analiz ve metabolik profil oluşturma

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

dissection-mouse-pancreas-for-histological-analysis-metabolic

Research

JoVE Journal

Medicine

36.2K Görüntüleme.  Miami University. Video bu makalede ayrıntılı bir gösteri diseksiyon histolojik analizi ve metabolik profil oluşturma için başarıyla bir fareden pankreas kaldırmak için gerekli yordamlar sağlar.

Video: Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

Dissection of Human Vitreous Body Elements for Proteomic Analysis

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

This video shows an effective technique for differentiating and dissecting the various semi-transparent structures of the human vitreous body in post mortem eyes.

Proteomik Analizi İnsan Vitreus Gövde Elemanları diseksiyonu

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions ...

Tıp

Collection Protocol for Human Pancreas

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation of pancreas recovered from cadaveric organ donors. The pancreas is divided into 3 main regions (head, body, tail) followed by serial transverse sections throughout the medial to lateral axis. Alternating sections are used for fixed paraffin and fresh frozen blocks and remnant samples are minced for snap frozen sample preparations, either with or without RNAse inhibitors, for DNA, RNA, or protein isolation. The overall goal of the pancreas dissection procedure is to sample the entire pancreas while maintaining anatomical orientation.
Endocrine cell heterogeneity in terms of islet composition, size, and numbers is reported for human islets compared to rodent islets 1. The majority of human islets from the pancreas head, body and tail regions are composed of insulin-containing &beta;-cells followed by lower proportions of glucagon-containing &alpha;-cells and somatostatin-containing &delta;-cells. Pancreatic polypeptide-containing PP cells and ghrelin-containing epsilon cells are also present but in small numbers. In contrast, the uncinate region contains islets that are primarily composed of pancreatic polypeptide-containing PP cells 2. These regional islet variations arise from developmental differences. The pancreas develops from the ventral and dorsal pancreatic buds in the foregut and after rotation of the stomach and duodenum, the ventral lobe moves and fuses with the dorsal 3. The ventral lobe forms the posterior portion of the head including the uncinate process while the dorsal lobe gives rise to the rest of the organ. Regional pancreatic variation is also reported with the tail region having higher islet density compared to other regions and the dorsal lobe-derived components undergoing selective atrophy in type 1 diabetes4,5.
Additional organs and tissues are often recovered from the organ donors and include pancreatic lymph nodes, spleen and non-pancreatic lymph nodes. These samples are recovered with similar formats as for the pancreas with the addition of isolation of cryopreserved cells. When the proximal duodenum is included with the pancreas, duodenal mucosa may be collected for paraffin and frozen blocks and minced snap frozen preparations.

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

İnsan Pankreas için Toplama Protokolü

This dissection and sampling procedure was developed for the Network for Pancreatic Organ Donors with Diabetes (nPOD) program to standardize preparation ...

Whole Vitreous Humor Dissection for Vitreodynamic Analysis

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous studies have shown the results of such dissections, the detailed steps have not been described, precluding researchers outside the field from replicating their methods. Other studies harvest vitreous either through aspiration, which does not maintain the vitreous structure anatomy, or through partial dissection, which only isolates the vitreous core. The proposed method isolates the whole vitreous body, with the vitreous core and cortex intact, while maintaining vitreous anatomy and structural integrity. In this method, a full thickness scleral flap in an enucleated porcine eye is first created and through this, the choroid tissue can be separated from the sclera. The scleral flap is then expanded and the choroid is completely separated from the sclera. Finally the choroid-retina tissue is peeled off the vitreous to leave an isolated intact vitreous body. The proposed vitreous dissection technique can be used to study physical properties of the vitreous humor. In particular, this method has significance for experimental studies involving drug delivery, vitreo-retinal oxygen transport, and intraocular convection.

The goal of this protocol is to show an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes.

Vitreodynamic Analiz Tüm Vitreus Mizah Diseksiyon

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous ...

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

This protocol describes an injury model involving the surgical ligation of the pancreatic duct in the adult mouse pancreas, resulting in severe injury that establishes an environment that allows beta cell neogenesis and proliferation. This model can be used as a tool to study mechanisms involved in beta cell formation.

Endokrin ve Ekzokrin yeniden programlanması ve Silahların Yayılmasını Önleme Modeli Olarak Pankreas kanalının bağlanmasını yoluyla Fare Pankreas Cerrahi Sakatlık

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Do's and Don'ts in the Preparation of Muscle Cryosections for Histological Analysis

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of neuromuscular disorders. However, in order to do so, proper care needs to be taken when excising and preserving tissues to achieve optimal staining. One method of tissue preservation involves fixing tissues in formaldehyde and then embedding them with paraffin wax. This method preserves morphology well and allows for long-term storage at RT but is cumbersome and requires handling of toxic chemicals. Further, formaldehyde fixation results in antigen cross-linking, which necessitates antigen retrieval protocols for effective immunostaining. On the contrary, frozen sectioning does not require fixation and thus retains biological antigen conformation. This method also provides a distinct advantage in quick turn around time, making it especially useful in situations needing quick histological evaluation like intraoperative surgical biopsies. Here we describe the most effective method of preparing muscle biopsies for visualization with different histological and immunological stains.

Here we demonstrate the most efficient methods for freezing, embedding, cryosectioning, and staining of muscle biopsies to avoid freezing artifacts.

Histolojik Analiz kas cryosections Hazırlanmasında Yapılması ve Yapılmaması Gerekenler

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

Fare modeli değiştirilmiş manşet tekniği kullanarak pankreas nakli için

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed in mice is high. Phenotypic characterization of these mouse models also requires the examination of cardiac function and morphology. Echocardiography and magnetic resonance imaging (MRI) are commonly used approaches to characterize cardiac function and morphology in mice. Echocardiographic and MRI equipment specialized for use in small rodents is&#160;expensive and requires a dedicated space. This protocol describes cardiac measurements in mice using a clinical echocardiographic system with a 15 MHz human vascular probe. Measurements are performed on anesthetized adult mice. At least three image sequences are recorded and analyzed for each animal in M-mode in the parasternal short-axis view. Afterwards, cardiac histological examination is performed, and cardiomyocyte diameters are determined on hematoxylin-eosin- or wheat germ agglutinin (WGA)-stained paraffin sections. Vessel density is determined morphometrically after Pecam-1 immunostaining. The protocol has been applied successfully to pharmacological studies and different genetic animal models under baseline conditions, as well as after experimental myocardial infarction by the permanent ligation of the left anterior descending coronary artery (LAD). In our experience, echocardiographic investigation is limited to anesthetized animals and is feasible in adult mice weighing at least 25 g.

Echocardiographic examination is frequently used in mice. Expensive high-resolution ultrasound devices have been developed for this purpose. This protocol describes an affordable echocardiographic procedure combined with histological morphometric analyses to determine cardiac morphology.

Kardiyak Morfoloji Ekokardiyografik ve histolojik muayene fare

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

Dissection and Explant Culture of Murine Allantois for the In Vitro Analysis of Allantoic Attachment

The placenta is essential for the growth and development of mammalian embryos. For this reason, numerous genetic alterations and likely also environmental insults that disturb placenta development or function can cause early pregnancy loss in mice and humans. Nevertheless, simple in vitro assays to screen for potential effects on placenta formation are lacking. Here, we focus on modeling the first and critical step in placenta formation, which consists of the attachment of the allantois to the chorion. We describe a method to rapidly assess the attachment of allantoic explants on immobilized &#945;4&#946;1 integrin, which serves as a chorio-mimetic substrate.This in vitro approach enables a qualitative evaluation of the attachment and spreading behavior of multiple allantois explants at different consecutive time points. The protocol may be used to investigate the effect of targeted mouse mutations, drugs, or various environmental factors that have been linked to pregnancy complications or fetal loss on allantois attachment ex vivo.

Dissection and Explant Culture of Murine Allantois for the In Vitro Analysis of Allantoic Attachment

We describe an in vitro assay to model chorioallantoic attachment, the first step in placenta formation. The protocol demonstrates the dissection and explant culture of murine allantoides on immobilized &#945;4&#946;1 integrin. Allantois attachment is evaluated microscopically at pre-determined time points.

Diseksiyon ve Explant kültür Allantoic eki Vitro analizi için fare Allantois ve

The placenta is essential for the growth and development of mammalian embryos. For this reason, numerous genetic alterations and likely also environmental ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

Aortopathy aort anevrizması β-aminopropionitrile-indüklenen ve diseksiyon, fare modeli nicel mikro-CT Analizi

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Mechanisms Underlying Gut Hormone Secretion Using the Isolated Perfused Rat Small Intestine

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, nutrient absorption and distribution, and post-prandial glucose excursions. Understanding the molecular mechanisms that regulate gut hormone secretion is fundamental for understanding and translating gut hormone physiology. Traditionally, the mechanisms underlying gut hormone secretion are either studied in vivo (in experimental animals or humans) or using gut hormone-secreting primary mucosal cell cultures or cell lines. Here, we introduce an isolated perfused rat small intestine as an alternative method for studying gut hormone secretion. The virtues of this model are that it relies on the intact gut, meaning that it recapitulates most of the physiologically important parameters responsible for the secretion in in vivo studies, including mucosal polarization, paracrine relationships and routes of perfusion/stimulus exposure. In addition, and unlike in vivo studies, the isolated perfused rat small intestine allows for almost complete experimental control and direct assessment of secretion. In contrast to in vitro studies, it is possible to study both the magnitude and the dynamics of secretion and to address important questions, such as what stimuli cause secretion of different gut hormones, from which side of the gut (luminal or vascular) is secretion stimulated, and to analyze in detail molecular sensors underlying the secretory response. In addition, the preparation is a powerful model for the study of intestinal absorption and details regarding the dynamics of intestinal absorption including the responsible transporters.

Here, we present a powerful and physiological model to study the molecular mechanisms underlying gut hormone secretion and intestinal absorption &#8212; the isolated perfused rat small intestine.

Gut hormonu salgılanması izole kullanarak temel mekanizmaları fare bağırsağı periosteum

The gut is the largest endocrine organ of the body, producing more than 15 different peptide hormones that regulate appetite and food intake, digestion, ...