The authors acknowledge the following funding sources, Whittier Foundation, Harrington Foundation, National Institutes of Health and Research to Prevent Blindness.

Tüm enükleasyon gözler bir mezbahaya elde edildi ve tüm deneyler kurumsal biyogüvenlik yasalarına uygun olarak yapıldı. <ol><li> Bir yüzey üzerinde enüklee gözü aşağı sabitleyin. Göz çevresinde aşırı doku yoluyla doku işaretçilerine yerleştirerek ve bir strafor tahta içine aşağı güvence bunu yapın. </li><li> Incelemek ve göz perilimbal konjonktivayı ayırın. <ol><li> Limbusta konjonktiva insizyon ve açık açık sklera kapalı incelemek için ince forseps (0.3 forseps) ve Microscissors (Westcott makas) kullanın. Künt diseksiyon daha diseksiyon izin ilerledikçe limbusun boyunca konjonktivayı kesin. </li><li> Mümkün olduğunca sklera açığa için her yolu göze (360 derece) etrafında konjonktivayı çıkarın. NOT: Bu prosedür geri kalanı boyunca, cerrahi iğne veya bir forseps ile göz fikse kolaylaştırmak için konjonktiva küçük bir miktar terk etmek yararlı olur. </li></ol></li><li> Biri boyunca tam kalınlıkta skleral flep oluşturmagöz (neşter bıçağı 69) yan. <ol><li> Yavaşça koyu pigmentli koroid kadar bir neşter (neşter bıçağı 11) ile sklera içine keserek limbusa limbusun ve 3 mm posterior bir ~ 5 mm skleral kesi paralel hale görünür. Koroid kendisi kesilirken için dikkatli olun. </li><li> Yavaşça içeri koroid iterek bu dokular arasındaki potansiyel boşluk büyütmek için mümkün kadar dikkatlice, (künt diseksiyon kullanarak) makasla veya neşter ile ya sklera ve koroid arasındaki düzlemi boyunca teşrih. </li><li> T-şekilli kesi oluşturarak, keskin microscissors ilk skleral kesi dik bir skleral kesi yapmak. </li><li> Sonra açık açık yatan koroid gelen skleral doku kaldırmak ve yavaşça yukarıda belirtildiği gibi sklera uzak koroid itmek için bir çevresel biçimde incelemek için devam ediyor. </li><li> Gerektiği gibi skleral flep büyütün. <ol><li> Yavaşça uzak skleradan koroid teşrih küntsüreci boyunca. </li><li> Limbusun dik yapılan kesi optik sinir ulaşır ve kesi limbusa paralel göz çevresi (45 °) en az üçte biri kadar yapılan flep büyütün. </li><li> Künt diseksiyon sırasında göz işlemek için kaldıraç kaynağı olarak skleral flep kullanın. kapağın karşı çekiş künt diseksiyon için kolaylaştırır. </li></ol></li><li> Diğer skleral flep önceki adımı yineleyin. </li></ol></li><li> Koroid geniş bir alana maruz skleral flep kesip. </li><li> Gözün çevresi (360 derece) etrafında 3. adımda yapılan kesi devam edin. </li><li> Kalan koroid-retina doku çıkarın. <ol><li> Maruz Bölgede, hafifçe geriye kalan koroid-retina dokusu savmak için bir pamuk-ucu kullanın. Alternatif olarak, hafifçe forseps ile doku kapmak ve altta yatan vitreus kapalı soyulması. </li><li> Gerekirse koroid st içine bir kesi yapmakoptik sinir Arting. Sonra retina ve vitreus korteksi maruz yavaşça koroid uzak soyun. </li></ol></li><li> Uzakta skleradan koroid diseksiyon künt Devam ve daha sonra tüm bozulmadan vitrözü elde etmek koroid uzak soyun. </li><li> İstenilen yerde bütün vitrözü konumlandırmak için korneaya bağlı skleral jant kullanın. Bu noktada, vitröz ön sklera ve mercek takılır. </li><li> Tüm sklera kaldırarak, lens etrafında, sklera içinden vitrözü teşrih künt. </li><li> Gerekirse uzak vitreus lensi kepçe künt bir alet kullanın. Çeşitli vitreodynamic deneyler için örnek kullanın. <ol><li> Havaya maruz kalan yüzey alanı, bilinen bir cam beher içinde örnek yerleştirin. Numunenin kenarında oksijen duyarlı probu yerleştirin. Örnek olarak bilinen bir mesafe (R) probu hareket ettirmek için, bir mikro-manipülatör kullanın. </li><li> Teorik difüzyon denklemini elde etmek için difüzyon Fick yasaları kullanın. Dekar ileta adımda 10.1 toplanan vb deneysel difüzyon katsayısı / reaksiyon süresini elde </li></ol></li></ol>

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The authors have nothing to disclose.

Dikkatle vitreus diseksiyonu sırasında yapılmalıdır iki kritik adımlar vardır. Tam kalınlıkta skleral flep oluşturma Adım 3, tüm diseksiyon için çok önemlidir. Bakım tam kalınlıkta skleral kanatçık oluştururken koroid halinde kesmek için önlem alınmalıdır. diğer kritik adım koroid gelen sklera uzak diseksiyon olduğunu. Bu adım dikkatle vitreus dışarı dökmek olabilir hangi koroid birden delikler oluşturarak önlemek için yapılmalıdır. Protokolü değiştirmek ve hala bozulmamış t?...

Bu yöntemin amacı, detay vitreodynamic analiz amaçları için, kadavra göz, camsı çekirdek ve korteks bozulmamış bir bütün dokunulmamış vitreus izole etmek için kullanılan bir tekniktir. Vitreus fizyoloji alanında böyle akışkanlar mekaniği araştırmacılar olarak, çok disiplinli araştırmacı, büyüdü gibi, vitreus 1 fiziksel ve biyomekanik özelliklerini araştırıyor. Bu amaçla, detaylara multi-disipliner araştırmacılar yardımcı olmak için bütün, sağlam camsı vücudu izole etmek için bir tekniktir esastır. Sebag ve ark., 2. ve diğerleri 3 insan kadavra göze şık bütün vitreus diseksiyonu yapılan ve sonuçları illüstrasyonlar gösterdi. Ancak, kullanılan teknik, bağımsız yöntemi çoğaltmak mümkün olmaz detay ve uzman olmayanların tarif edilmemiştir. Diğer çalışmalar, böyle bir aspirasyon veya kısmi diseksiyonu gibi basit yöntemler kullanılarak kadavra gözlerinden vitreus hasat varher ikisi de bütün dokunulmamış vitreus neden yoktur. Gisladottis ve ark., 4 ve Xu ve ark. 5 kadavra gözlerinden hasat vitreus içinde geçirgenliği araştırmak. Camsı ekstraksiyon hiç bir yöntem tarif edilmiştir Bununla birlikte, bu bir şırınga ile vitreus aspire varsayılmıştır. Watts ve ark., 6 cerrahi teknikle tavşan vitreus mizah izole bir yöntem anlatarak bir adım daha ileri gitti. Bununla birlikte, bu yöntem sadece camsı çekirdek bir yalıtım olup camsı korteks ile sonuçlanır. Skeie ve ark., 7 sonra 4 eşsiz bölgelere vitrözü organize ve zarif analiz için her bölümünü incelemek için bir yöntem tarif. Bu teknik, bununla birlikte, bir bütün olarak, bozulmamış bir vitröz ile sonuçlanmaz. Geçerli bir teknik şu anda sadece kadavra gözlerinde yapılır biyofizik deneyler kolaylaştırmak için geliştirilmiştir. Önceki yöntem olarak tarif edilen birBove, 1) hiçbiri tamamen bütün vitreus vücudu izole çünkü sınırlıdır, 2) hasat vitreus çekirdek ve korteks 3) vitreus anatomik yapısı korunur değil, homojenize, ya da 4) diseksiyon teknikleri yeterince diğer alanlardaki araştırmacılar tarafından çoğaltma için ayrıntılı değil . Buna ek olarak, sklera ve koroid, vitreus görselleştirme opaklığına nedeniyle sağlam gözün sınırlıdır. Bütün bu gözün içine yapılabilir ölçümlerin hassasiyetini ve fizibilite sınırlar. Buna ek olarak, vitrözü çevreleyen anatomik yapılar vitreus biyokimyasal ve fiziksel özelliklerinin çalışma karıştırabilir. Son yıllarda, camsı bilimin vücut müthiş büyüdü ve tüm vitreus bireysel parçaları farklı özelliklere sahip olduğuna inanmak için bir neden yoktur. Vitreodynamics Araştırma, için vitreus fiziksel biyomekanik ve kimyasal özellikleri araştıran artan bir ilgi vardırBöyle ilaç dağıtım, intravitreal oksijenlenme 8 ve vitrektomi gibi klinik tıpta uygulamaları vardır h. Vitrözü işlemek için farmakolojik ajanlar kullanan Farmakolojik vitreodynamics, vitrektomi sonuçları 9 iyileştirmek için kullanılabilir. Biyomekanik özellikleri intravitreal ilaç verme teknolojileri 10-12 geliştirmek için kullanılabilir camsı sıvı akışı, modellemek için kullanılır. Vitreus çeşitli kesimleri fiziksel özellikleri vitro-retinal oksijen taşıma 13 anlaşılması açısından büyük önem taşıyor. Önerilen vitreus diseksiyon tekniği bozulmadan vitreus çeşitli özelliklerini incelemek için kullanılabilir. Bu tezgah üstü deneyleri daha iyi görselleştirme ile tüm, sağlam camsı organları yapılması mümkün kılar. Özet olarak, vitröz çalışma için mevcut yöntemler ya yeterince tarif değildir, ya da eksik bir izolasyon camsı nüve ve korteks ile sonuçlanır. Bu nedenle, e gerçekleştirmek için bir ihtiyaç vardırşeffaf göz modelinde xperiments kadavra gözün var vitreus anatomisi korurken.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>0.3 forceps</td>
			<td>Storz Opthalmics</td>
			<td>E1793</td>
			<td></td>
		</tr>
		<tr>
			<td>Westcott Tenotomy Scissors Curved Right</td>
			<td>Storz Opthalmics</td>
			<td>E3320 R</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Handle No. 3</td>
			<td>VWR</td>
			<td>25607-947</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Blade, #11, for #3 Handle</td>
			<td>VWR</td>
			<td>470174-844</td>
			<td></td>
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</table>

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.

Protokol sonrasında sağlam çekirdek ve korteks (Şekil 3) ile başarılı bir vitreus diseksiyon yol açacaktır. Bu vitröz korteks yapıştırılmış retina kalıntı parçalar bellidir. Bozulmamış tam vitröz hümör, belirli vitreodynamic deneyler için çeşitli şekillerde kullanılabilir. Bizim durumumuzda, difüzyon sağlam vitreus oksijen oranı ve karşılık gelen zaman sabiti (Şekil 2) incelendi var. Bizim yöntemi kullanılarak (çekirdek ve korteks) ayrılmıştır ...

Watch this Scientific Journal Video about Whole Vitreous Humor Dissection for Vitreodynamic Analysis at JoVE.com

Whole Vitreous Humor Dissection for Vitreodynamic Analysis

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 ...

whole-vitreous-humor-dissection-for-vitreodynamic-analysis

Research

JoVE Journal

Medicine

11.3K Görüntüleme.  University of Southern California. 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.

Video: Whole Vitreous Humor Dissection for Vitreodynamic Analysis

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

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

Farelerde Uzun Süre Whole Body Hipertermi için protokol

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

3D Whole-heart Myocardial Tissue Analysis

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other biologicals that enhance angio- or vasculogenesis into the infarct border zone (IBZ), tissue perfusion is improved, and the myocardium can be protected from further damage. For maximum therapeutic effect, it is hypothesized that the regenerative substance is best delivered to the IBZ. This requires accurate injections and has led to the development of new injection techniques. To validate these new techniques, we have designed a validation protocol based on myocardial tissue analysis. This protocol includes whole-heart myocardial tissue processing that enables detailed two-dimensional (2D) and three-dimensional (3D) analysis of the cardiac anatomy and intramyocardial injections. In a pig, myocardial infarction was created by a 90-min occlusion of the left anterior descending coronary artery. Four weeks later, a mixture of a hydrogel with superparamagnetic iron oxide particles (SPIOs) and fluorescent beads was injected in the IBZ using a minimally-invasive endocardial approach. 1 h after the injection procedure, the pig was euthanized, and the heart was excised and embedded in agarose (agar). After the solidification of the agar, magnetic resonance imaging (MRI), slicing of the heart, and fluorescence imaging were performed. After image post-processing, 3D analysis was performed to assess the IBZ targeting accuracy. This protocol provides a structured and reproducible method for the assessment of the targeting accuracy of intramyocardial injections into the IBZ. The protocol can be easily used when the processing of scar tissue and/or validation of the injection accuracy of the whole heart is desired.

This protocol describes a novel method for the 3D comparison of whole-heart myocardial tissue with MRI. This is designed for the accurate assessment of intramyocardial injections in the infarct border zone of a chronic porcine model of myocardial infarction.

3D Tüm kalp Miyokard Doku Analizi

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

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 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 ...

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&#8212;Application in Premanifest Huntington's Disease

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy begins many years prior to the motor onset (during the &#34;premanifest&#34; period), but the spatiotemporal pattern of regional atrophy across the brain has not been fully characterized. Here we demonstrate an online cloud-computing platform, &#34;MRICloud&#34;, which provides atlas-based whole-brain segmentation of T1-weighted images at multiple granularity levels, and thereby, enables us to access the regional features of brain anatomy. We then describe a regression model that detects statistically significant inflection points, at which regional brain atrophy starts to be noticeable, i.e. the &#34;change-point&#34;, with respect to a disease progression index. We used the CAG-age product (CAP) score to index the disease progression in HD patients. Change-point analysis of the volumetric measurements from the segmentation pipeline, therefore, provides important information of the order and pattern of structural atrophy across the brain. The paper illustrates the use of these techniques on T1-weighted MRI data of premanifest HD subjects from a large multicenter PREDICT-HD study. This design potentially has wide applications in a range of neurodegenerative diseases to investigate the dynamic changes of brain anatomy.

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&amp;#8212;Application in Premanifest Huntington&#039;s Disease

This paper describes a statistical model for volumetric MRI data analysis, which identifies the &#34;change-point&#34; when brain atrophy begins in premanifest Huntington&#39;s disease. Whole-brain mapping of the change-points is achieved based on brain volumes obtained using an atlas-based segmentation pipeline of T1-weighted images.

Bütün beyin segmentasyon ve değişim noktası analizi anatomik beyin MR'ı — uygulama Premanifest Huntington hastalığı

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy ...

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.

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

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 ...

Imaging and Analysis of Oil Red O-Stained Whole Aorta Lesions in an Aneurysm Hyperlipidemia Mouse Model

Apolipoprotein E (Apoe)- or low density lipoprotein receptor (Ldlr)-deficient hyperlipidemic mice are the two most commonly used models for atherosclerosis research. They are used to study the impact of a various genetic factors and different cell types on atherosclerotic lesion formation and as well as test the development of new therapies. Isolation, excision of the whole aorta, and quantification of Oil Red O-stained atherosclerotic lesions are basic morphometric methods used to evaluate atherosclerotic burden. The goal of this protocol is to describe an optimized, step-by-step surgical method to dissect, perfuse-fix, isolate, stain, image and analyze atherosclerotic lesions in mouse aortas with Oil Red O. Because atherosclerotic lesions can form anywhere in the entire aortic tree, this whole aorta Oil Red O staining method has the advantage of evaluating lipid-laden plaques in the entire aorta and all branches in a single mouse. In addition to Oil Red O staining, fresh isolated whole aortas can be used for variety of in vitro and in vivo experiments and cell isolations.

This protocol provides a step-by-step procedure to analyze atherosclerotic burden in mice. Investigators can use this protocol to compare the abundance, location, and size of atherosclerotic lesions in different animals.

Anevrizma Hiperlipidemi Fare Modelinde Yağ Kırmızı O-Boyalı Bütün Aort Lezyonlarının Görüntülenmesi ve Analizi

Apolipoprotein E (Apoe)- or low density lipoprotein receptor (Ldlr)-deficient hyperlipidemic mice are the two most commonly used models for ...

Location, Dissection, and Analysis of the Murine Stellate Ganglion

The autonomic nervous system is a substantial driver of cardiac electrophysiology. Especially&#160;the role of its sympathetic branch is an ongoing matter of investigation in the pathophysiology of ventricular arrhythmias (VA). Neurons in the stellate ganglia (SG)&#160;&#8211;&#160;bilateral star-shaped structures of the sympathetic chain &#8211;&#160;are an important component of the sympathetic infrastructure. The SG are a recognized target for&#160;treatment via cardiac sympathetic denervation in patients with therapy-refractory VA. While neuronal remodeling and glial activation in the SG have been described in patients with VA, the underlying cellular and molecular processes that potentially precede the onset of arrhythmia are only insufficiently understood and should be elucidated to improve autonomic modulation. Mouse models allow us to study sympathetic neuronal remodeling, but identification of the murine SG is challenging for the inexperienced investigator. Thus, in-depth cellular and molecular biological studies of the murine SG are lacking for many common cardiac diseases. Here, we describe a basic repertoire for dissecting and studying the SG in adult mice for analyses at RNA level (RNA isolation for gene expression analyses, in situ hybridization), protein level (immunofluorescent whole mount staining), and cellular level (basic morphology, cell size measurement). We present potential solutions to overcome challenges in the preparation technique, and how to improve staining via quenching of autofluorescence. This allows for the visualization of neurons as well as glial cells via established markers in order to determine cell composition and remodeling processes. The methods presented here allow characterizing the SG to gain further information on autonomic dysfunction in mice prone to VA and can be complemented by additional techniques investigating neuronal and glial components of the autonomic nervous system in the heart.

Pathophysiological changes in the cardiac autonomic nervous system, especially in its sympathetic branch, contribute to the onset and maintenance of ventricular arrhythmias. In the present protocol, we show how to characterize murine stellate ganglia to improve the understanding of the underlying molecular and cellular processes.

Murine Stellate Ganglion'un Yeri, Diseksiyonu ve Analizi

The autonomic nervous system is a substantial driver of cardiac electrophysiology. Especially&#160;the role of its sympathetic branch is an ongoing matter ...

A Non-Invasive Method for Generating the Cyclic Loading-Induced Intra-Articular Cartilage Lesion Model of the Rat Knee

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely correlated with a history of articular cartilage damage and ligament avulsion. Surgical animal models of OA of the knee play an important role in understanding the onset and progression of post-traumatic OA and aid in the development of novel therapies for this disease. However, non-surgical models have been recently considered to avoid traumatic inflammation that could affect the evaluation of the intervention. 
In this study, an intra-articular cartilage lesion rat model induced by in vivo cyclic compressive loading was developed, which allowed researchers to (1) determine the optimal magnitude, speed, and duration of load that could cause focal cartilage damage; (2) assess post-traumatic spatiotemporal pathological changes in chondrocyte vitality; and (3) evaluate the histological expression of destructive or protective molecules that are involved in the adaptation and repair mechanisms against joint compressive loads. This report describes the experimental protocol for this novel cartilage lesion in a rat model.

Here, we present the cyclic loading-induced intra-articular cartilage lesion model of the rat knee, generated by 60 cyclic compressions over 20 N, resulting in damage to the femoral condylar cartilage in rats.

Sıçan Dizinin Döngüsel Yüklemeye Bağlı Eklem İçi Kıkırdak Lezyon Modelini Oluşturmak için Non-İnvaziv Bir Yöntem

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely ...