기금은 시력을 위해 전투에 의해 제공되었다. 조직은 아이오와 라이온스 안구 은행에서 얻은되었습니다.

1. 앞쪽에 세그먼트 해부. <ol><li> 각막은 먼저 15 ° 블레이드 (그림 1)를 사용하여 앞쪽에 챔버에 limbus에서 절개하여 제거됩니다. 다음 곡선 각막 - scleral 가위 중 하나 블레이드는 앞쪽에 챔버에 삽입됩니다. 서컴프렌셜 인하는 limbus 단 앞쪽에, 각막에 만들어집니다. </li><li> 0.12 콜리 브리 포셉 및 웨스트컷 가위는 모양체에 circumferentially 앞쪽에 아이리스 절단하는 데 사용됩니다. </li><li> 렌즈 핵은 0.12 콜리 브리의 포셉 또는 15 ° 블레이드 (supersharp)와 포셉와 피질과 캡슐로 제거됩니다. </li></ol> 2. 유리체 핵심 흡인. <ol><li> 5 CC의 주사기에 23 게이지 주사 바늘을 삽입하는 중간 유리 (그림 1). </li><li> 약 유리체 코어 1 ML 이상 부드럽게 aspirated입니다. </li><li> 예제는 다음 microfuge 튜브에서 다음 액체 질소에 배치됩니다. </li></ol> 3. 앞쪽에 Hyaloid의 해부. <ol><li> 앞쪽에 hyaloid은 구스 넥 현미경 조명의 입사 광을 조정하여 반투명 링으로 여겨진다. 유리체 핵심 이전 열망은 쉽게 식별을위한 다른 구조에서 앞쪽에 hyaloid를 분리합니다. </li><li> 탄성 조직의 시트는 신중하게 콜리 브리 또는 드레싱 포셉와 모양체에서 끌려나와 Vannas 가위로 절단됩니다. 이 책략이 반복됩니다. </li><li> 예제는 다음 microfuge 튜브에서 다음 액체 질소에 배치됩니다. </li></ol> 4. 유리체베이스 해부. <ol><li> 아이 컵을 안정화시키기 위해 0.12 집게를 사용하여 웨스트컷 가위는 시신경의 머리를 향해 모양체에서 네 개의 동등 간격 스트레스 덜어 인하함으로써 &quot;꽃&quot;눈에하는 데 사용됩니다. </li><li> 모양체 (그림 2)의 갈 거예요의 plicata는 웨스트컷 가위를 사용하여 quadrants 각에서 제거됩니다. </li><li> 유리체 기지를 다음 갈 거예요 플라나과 망막 (그림 2)을 통해 포셉로 오라 세라타의 양쪽에 파악됩니다. </li><li> 유리체 기지에 지속적인 견인이 집게와 함께 적용됩니다. 가 excised 그대로 웨스트컷 가위로 연속 절삭은 &quot;진주의 문자열&quot;모양과 반투명 조직을 추출합니다. </li><li> 예제는 다음 microfuge 튜브에서 다음 액체 질소에 배치됩니다. </li></ol> 5. 유리체 접속 제거. <ol><li> 갈 거예요 플라나은 오라 세라타 (그림 2) 사후 약 3 mm 전단지를 절단하여 웨스트컷 가위로 quadrants의 각 excised 수 있습니다. </li><li> 조직 전단지 사이에 유리체 피질은 세미 투명 필름 시각이다. </li><li> 탄성이 영화는 중 0.12 포셉로 파악 또는 샌드위치 - 셀 수술 스펀지을 준수하여 개최됩니다. 유리체 피질은 다음 떨어진 망막에서 가져온 및 웨스트컷 가위 (그림 1) excised 수 있습니다. </li><li> 샘플은 microfuge 튜브에서 다음 액체 질소에 배치됩니다. 모든 샘플은 실험을 위해 활용 -80 ° C까지에 저장됩니다. </li></ol> 6. 대표 결과 조직 샘플은 특정 실험을위한 다양한 방법으로 처리할 수 있습니다. 우리의 경우, 샘플은 SDS - PAGE (그림 3)에 의해 단백질 분석에 제출했다. <img src="/files/ftp_upload/2455/2455fig1.jpg" alt="그림 1" /> 그림 1. 인간의 눈의 횡단면보기 유리체 신체의 다른 substructures을 묘사한. 가장 앞쪽에 유리가 얇은 collagenous 층 앞쪽에 hyaloid라고합니다. 유리체 코어는 유리 몸 전체 중앙 지역을 구성되어 있습니다. 유리체의이 부분은 집게로 파악되어야하고 단단히 기본 모양체와 망막에 첨부되어 충분한 점성있는 유리 기지에 대비 더 수성이다. 유리체 코어를 포함한 것은 유리체 피질이라고하는 매우 얇은 껍질 collagenous입니다. <img src="/files/ftp_upload/2455/2455fig2.jpg" alt="그림 2" /> 그림 2. 유리체 기본 해부학. 유리체베이스 모양체와 망막을 분리 나누어 줄 수있는 오라 세라타 (흰색 화살표)를 따라 위치한 유리 신체의 반투명 지하입니다. 유리체 기지의 앞쪽에 경계선은 모양체의 갈 거예요 플라나 (흰색 라인)을 통해 확장됩니다. 유리체 기지의 사후 경계선 오라 세라타 (흰색 대쉬)로 사후 2 - 3 - mm를 확장합니다. 소비세에 유리 기지, 포셉는 조직을 파악하고 기본 모양체와 망막에서 멀리 당겨하는 데 사용됩니다. 일단 고가, 웨스트컷 가위는 기지를 따라 절단하는 데 사용됩니다. <img src="/files/ftp_upload/2455/2455fig3.jpg" alt="그림 3" /> 그림 3. 앞쪽에 hyaloid에 대한 한 차원 유리 바디 요소의 SDS - PAGE. 총 단백질 농도, 유리 기지, VITreous 코어 및 유리체 피질은 각각 11.24, 20.1, 16.61, 14.24 MG / ML,했다. 젤 전기 영동은 플라밍고 (바이오 래드) 물들일, 45 분 200 KV에서 수행하고 VersaDoc 이미징 시스템 (생물 래드)를 사용하여 시각되었습니다. 다른 조직에 대한 프로파일 differentially화된 단백질을 나타내는 (별표) 보존 또는 단백질 교차 오염뿐만 아니라, 독특한 밴드 중 하나를 나타내는 몇 가지 유사한 밴드를 보여줍니다.

<ol>
	<li>Bishop, P. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+macromolecules+and+supramolecular+organisation+of+the+vitreous+gel.">Structural macromolecules and supramolecular organisation of the vitreous gel.</a> Prog Retin Eye Res. 19, 323-344 (2000).</li><li>Sebag, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+biology+of+pharmacologic+vitreolysis.">Molecular biology of pharmacologic vitreolysis.</a> Trans Am Ophthalmol Soc. 103, 473-494 (2005).</li><li>Yoshimura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+analysis+of+inflammatory+immune+mediators+in+vitreoretinal+diseases.">Comprehensive analysis of inflammatory immune mediators in vitreoretinal diseases.</a> PLoS One. 4, e8158-e8158 (2009).</li><li>Gao, B. B., Chen, X., Timothy, N., Aiello, L. P., Feener, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+vitreous+proteome+in+diabetes+without+diabetic+retinopathy+and+diabetes+with+proliferative+diabetic+retinopathy.">Characterization of the vitreous proteome in diabetes without diabetic retinopathy and diabetes with proliferative diabetic retinopathy.</a> J Proteome Res. 7, 2516-2525 (2008).</li><li>Izuta, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+SOD+and+VEGF+are+increased+in+vitreous+bodies+from+proliferative+diabetic+retinopathy+patients.">Extracellular SOD and VEGF are increased in vitreous bodies from proliferative diabetic retinopathy patients.</a> Mol Vis. 15, 2663-2672 (2009).</li><li>Azimzadeh, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formalin-fixed+paraffin-embedded+(FFPE)+proteome+analysis+using+gel-free+and+gel-based+proteomics.">Formalin-fixed paraffin-embedded (FFPE) proteome analysis using gel-free and gel-based proteomics.</a> J Proteome Res. 9, 4710-4720 (2010).</li></ol>

관심 없음 충돌 선언하지 않습니다.

유리체베이스, 코어, 피질, 그리고 앞쪽에 hyaloid : 유리 몸은 누구의 분자 구성이 저조한 특히 substructures의 수준에서 이해 반투명 젤입니다. 유리체 핵심은 당뇨 망막 병증과 같은 질환과 연관된되었습니다 콘드로이틴 황산염의 proteoglycans, heparan의 황산염의 proteoglycans와 유리체 핵심에 hyaluronan. 1,2 단백질 생체와 함께, 콜라겐으로 합성한 II, V, IX, 그리고 XI가 포함되어 있습니다. 3-5 어떻게 이러한 단백질은 differentially substructures 각 표현하고 있으며, 많은 경우에 특정 단백질 신원은 알려져 있지 않습니다. 이러한 내용은 특정 vitreoretinal 질환과 관련된 단백질의 기원에 대한 통찰력을 제공하고 대상 미래 요법을하는 데 도움이 될 수 있습니다. 조직의 절개에 대한 최적의 해부 간격이 결정되지 않았지만, 단백질이 저하 하류 실험에 영향을 미칠 수 있습니다. 예를 들어, immunohistochemistry 12 시간 사후 눈에 영향과 일부 특정 효소 활동 (관찰되지 않은) 몇 시간 이내에 줄어들 수 있습니다. 이 연구의 모든 조직은 proteomics의 analsyis을위한 단백질 표현이나 적합성에 큰 변화없이 사망 2, 8 시간 사이에 수집되었다. 보존의 액체 질소 동결 방법이 LC-MS/MS 다른 조직에서 증명되었습니다 정착액 상호 연결로 인한 단백질 구조의 작은 변화를 방지하기 위해 고정 이상 선택합니다. 6 Proteomic 연구 정확하게하는 능력에 따라 달라집니다 본 동영상 실험에서 보여주 유리체의 다른 구획을 해부하다. 우리는 1 차원 SDS - PAGE를 사용하여 절개 기술을 입증했습니다. 우리의 결과는 제안으로, differentially 다양한 유리 바디 substructures에 단백질이 표현됩니다. 이러한 단백질을 확인하는 것이 유리 compartmentalization의 자세한 이해를 제공합니다.

<table border="1"><tbody><tr><td> 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> 0.12 포셉 </td><td> Storz Ophthalmics </td><td> E1502 </td></tr><tr><td> 5 CC 주사기 </td><td> 백톤 디킨슨 - </td><td> 309603 </td></tr><tr><td> 톱니 모양의 스트레이트 드레싱 포셉 </td><td> Storz Ophthalmics </td><td> E1400 </td></tr><tr><td> 23 게이지 바늘 </td><td> 백톤 디킨슨 - </td><td> 305145 </td></tr><tr><td> 콜리 브리 포셉 </td><td> Storz Ophthalmics </td><td> 132분의 2 </td></tr><tr><td> Castroviejo 직각 각막 가위 </td><td> Storz Ophthalmics </td><td> E3223 </td></tr><tr><td> Vannas 곡선 Capsulotomy 가위 </td><td> Storz Ophthalmics </td><td> E3387 </td></tr><tr><td> 샌드위치 - 셀 외과 스피어스 </td><td> 메드 트로닉 </td><td> 0,008,680 </td></tr><tr><td> 웨스트컷 곡선 Tenotomy 가위 </td><td> Storz Ophthalmics </td><td> E3320 </td></tr></tbody></table>

dissection of human vitreous body elements for proteomic analysis

유리체는 안구와 overlies 망막의 내부를 채우고 광학 명확 collagenous 세포외 기질이다. 유리체 substructures와 망막 사이에 1,2 비정상적인 상호 작용은 망막 눈물과 분리, 황반 주름, 황반 구멍 등 여러 vitreoretinal 질환, 기초 연령 관련 황반 변성, vitreomacular 트랙션, proliferative vitreoretinopathy, proliferative 당뇨성 망막 병증의 및 상속 vitreoretinopathies. 1,2 유리체 substructures의 분자 구성을 알 수 없습니다. 유리체 시체가 제한된 수술 액세스할 수있는 투명하기 때문에, 그것은 분자 수준에서 그 substructures을 공부하기 어려운되었습니다. 우리는 proteomic 및 생화 학적 분석을 위해 이러한 조직을 분리하여 보존하는 방법을 개발했습니다. 이 실험 동영상 해부 기술은 유리 기지 앞쪽에의 hyaloid, 유리 코어, 그리고 사후에 인간의 눈에서 유리 피질을 분리하는 방법을 보여줍니다. 각 유리체 구성 요소 중 하나는 차원 SDS - PAGE 분석은 우리의 해부 기술은 인간의 유리 본체의 각 기초 공사에 해당하는 사 고유의 단백질 프로파일 결과 것으로 나타났다. differentially들에게만 단백질의 식별이 후보 분자는 다양한 vitreoretinal 질병을 기본 공개합니다.

Watch this Scientific Journal Video about Dissection of Human Vitreous Body Elements for Proteomic Analysis at JoVE.com

Dissection of Human Vitreous Body Elements for Proteomic Analysis

이 동영상은 사후의 눈에 인간의 유리 신체의 다양한 반투명 구조를 차별하고 해부를위한 효과적인 기법을 보여줍니다.

Proteomic 분석을위한 인적 유리 바디 요소의 해부

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

dissection-of-human-vitreous-body-elements-for-proteomic-analysis

Research

JoVE Journal

Medicine

31.7K 조회수.  University of Iowa. 이 동영상은 사후의 눈에 인간의 유리 신체의 다양한 반투명 구조를 차별하고 해부를위한 효과적인 기법을 보여줍니다.

비디오: Dissection of Human Vitreous Body Elements for Proteomic Analysis

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 분석을위한 항목의 유리체 유머 해부

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

연구

의학

Laser-capture Microdissection of Human Prostatic Epithelium for RNA Analysis

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types. Healthy prostate is comprised of fibromuscular stroma surrounding discrete epithelial-lined secretory lumens and a very small population of immune and neuroendocrine cells. In contrast, areas of prostate cancer have increased dysplastic luminal epithelium with greatly reduced or absent stromal population. Given the profound difference between stromal and epithelial cell types, it is imperative to separate the cell types for any type of downstream molecular analysis. Despite this knowledge, the bulk of gene expression studies compare benign prostate to cancer without micro-dissection, leading to stromal bias in the benign samples. Laser-capture micro-dissection (LCM) is an effective method to physically separate different cell types from a specimen section. The goal of this protocol is to show that RNA can be successfully isolated from LCM-collected human prostatic epithelium and used for downstream gene expression studies such as RT-qPCR and RNAseq.

The goal of this protocol is to use laser-capture micro-dissection as an effective method to isolate pure populations of cell types from heterogeneous prostate tissues for downstream RNA analysis.

RNA 분석을위한 인간의 전립선 상피 세포의 레이저 캡처 이외에 Microdissection

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types.  Healthy prostate is comprised of ...

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.

조직학 분석 및 대사 프로 파일링에 대 한 마우스 췌의 해 부

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.

해 부와 Allantoic 첨부 파일의 생체 외에서 분석 Murine Allantois Explant 문화

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

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

신체 구성 및 높은 지방 연 준 쥐에서 대사 배우면 분석

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

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.

Β-aminopropionitrile-유도 대동맥 동맥 및 해 부의 마우스 모델에서 Aortopathy의 양적 마이크로 CT 분석

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

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Segmentation and Linear Measurement for Body Composition Analysis using Slice-O-Matic and Horos

Body composition is associated with risk of disease progression and treatment complications in a variety of conditions. Therefore, quantification of skeletal muscle mass and adipose tissues on Computed Tomography (CT) and/or Magnetic Resonance Imaging (MRI) may inform surgery risk evaluation and disease prognosis. This article describes two quantification methods originally described by Mourtzakis et al. and Avrutin et al.: tissue segmentation and linear measurement of skeletal muscle. Patients' cross-sectional image at the midpoint of the third lumbar vertebra was obtained for both measurements. For segmentation, the images were imported into Slice-O-Matic and colored for skeletal muscle, intramuscular adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue. Then, surface areas of each tissue type were calculated using the tag surface area function. For linear measurements, the height and width of bilateral psoas and paraspinal muscles at the level of the third lumbar vertebra are measured and the calculation using these four values yield the estimated skeletal muscle mass. Segmentation analysis provides quantitative, comprehensive information about the patients' body composition, which can then be correlated with disease progression. However, the process is more time-consuming and requires specialized training. Linear measurements are an efficient and clinic-friendly tool for quick preoperative evaluation. However, linear measurements do not provide information on adipose tissue composition. Nonetheless, these methods have wide applications in a variety of diseases to predict surgical outcomes, risk of disease progression and inform treatment options for patients.

Segmentation and linear measurements quantify skeletal muscle mass and adipose tissues using Computed Tomography and/or Magnetic Resonance Imaging images. Here, we outline the use of Slice-O-Matic software and Horos image viewer for rapid and accurate analysis of body composition. These methods can provide important information for prognosis and risk stratification.

슬라이스-오-매틱 및 호로스를 사용하여 신체 조성 분석을 위한 세분화 및 선형 측정

Body composition is associated with risk of disease progression and treatment complications in a variety of conditions. Therefore, quantification of ...

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.

뮤린 스텔레이트 갠리온의 위치, 해부 및 분석

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.

래트 무릎의 순환 로딩 유도 관절내 연골 병변 모델을 생성하기 위한 비침습적 방법

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