The authors would like to thank the Lam Lab at Georgia Tech for many helpful discussions in developing the experimental assays. Research reported in this publication was supported by the National Heart, Lung, and Blood Institute of the National Institutes of Health under Award Number K01HL115486 and by New Innovator Grant 1DP2OD007433-01 from the Office of the Director, National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Not: Aşağıdaki protokol Georgia Tech Kurumsal Değerlendirme Kurulu (KİK) tarafından belirlenen kurallara uyar. 1. Kan Toplama ve Eritrosit İzolasyon Prosedürü <ol><li> 10 ml heparinli boşaylı tüpler içinde bağışçılardan kan 40-120 ml toplayın. Toplama 4 saat içinde PBMC (periferal kan mononükleer hücre) izolasyon başlatın. Bu süre boyunca, oda sıcaklığında kan tutun. NOT: Bu alt PBMC verim neden olacak gibi RT ya da 4 ° C kan O / N depolama tavsiye edilmez. </li><li> Tam kan 1 seyreltin: 1 buz soğukluğundaki (4 ° C) steril PBS içinde. </li><li> Pipet boş bir steril 50 ml konik bir tüp içine buz soğukluğunda hidrofilik polisakarit 10 mi. Yavaşça hidrofilik polisakaritin üzerine seyreltilmiş tam kan aktarın. </li><li> Çok yavaş tüpün kenarı boyunca yavaş ayarı ve pipet kanda 25 ml&#39;lik pipet kullanın. Kan CENTRI önce sakkaritte ile karışık olmadığından emin olunhidrofilik polisakkaridler, aksi hücreleri ve toksik olan fugation için PBMC verimi daha düşük olacaktır. </li><li> 4 ° C&#39;de 400 x g&#39;de 30 dakika boyunca santrifüj kan örneği alındı. Varsa, santrifüj sonra daha iyi ayrılması için maksimum yarısına santrifüj fren hızını azaltır. </li><li> Santrifüj kan örneğinin plazma / trombosit fraksiyonu kapalı aspire. Dikkatle doğrudan paketlenmiş RBC katman üzerinde hidrofilik polisakkarit tabakası aspire. </li><li> Paketlenmiş RBCs 8-10 ml toplayın ve 1 sulandırmak: steril olmayan% 0.9 NaCl serum fizyolojik ile 1. </li></ol> 2. Konfokal Mikroskopi Değerlendirme Glike ve pıhtı Yapıları (Simüle Diyabetik pıhtısı) <ol><li> 37 ° C &#39;de, insan fibrinojen ve floresan etiketli insan fibrinojen konjugatı defrost. </li><li> Pipet 0.5 ml aşağıdaki mikrosantrifüj tüp mezun oldu. <ol><li> Sağlıklı normal fibrinojen pıhtılaşması için,% 10 floresan etiketli insan fibrinojen insan fibrinojen karıştırın5 mg / ml&#39;lik bir konsantrasyonda 50 mM Tris / 100 mM NaCI tampon maddesi içinde konjugat. </li><li> Yapay glikozile fibrinojen için, (diyabetik pıhtıları gibi örneğin) 6.8 mg / ml&#39;lik bir konsantrasyon elde edilen 50 mM Tris / 100 mM NaCI içinde çözülmüş glikoz 5 mM bir çözeltisi içindeki insan fibrinojen ve% 10 floresan etiketli fibrinojen konjugatı inkübe edilir. </li></ol></li><li> Işığa maruz kalmasını önlemek için bir opak malzeme kullanılarak mikro-santrifüj tüplerine örtün. 48 saat süre ile 37 ° C su banyosu içinde inkübe edin. </li><li> Sürgünün 2 tarafı üzerinde ince bir yapışkan sabitlenmesiyle, bir cam mikroskop lamı üzerine bir bölme yapın. </li><li> 48 saatlik bir bekletme süresinden sonra, oda sıcaklığında FXIIIa ve insan alfa trombini çözülüp, fibrin oluşumu için reaktifler hazırlamak. </li><li> 5 mM CaCl2 tampon içinde 80 U / ml FXIIIa seyreltin. </li><li> 5 mM CaCl2 tamponu içinde 4 U / ml trombin seyreltin. <ol><li> Normal pıhtılar için, fibrinojen, FXIIIa ve trombinin nihai konsantrasyonlar 2.5 m sağlamakg / ml, 20 U / ml, ve sırasıyla 50 ul bir hacimde, 1 U / ml olmuştur. </li><li> Glikozillenmiş pıhtıları için, fibrinojen FXIIIa ve trombin nihai konsantrasyonları sırası ile 50 ul&#39;lik bir hacimde, 3.4 mg / ml, 20 U / ml ve 1 U / ml olduğundan emin olun. </li></ol></li><li> Hemen yapışkanın oluşturduğu odasına fibrin çözeltisi 30 ul pipetle. </li><li> Yapışkan 2 kat üstüne 0.15 mm&#39;lik kalınlığa sahip bir 22 mm x 22 mm ebadındaki bir cam kapak kayma yerleştirin. Kurumasını fibrin pıhtısı önlemek için açık yapıştırıcı ile açık yanlarını Seal. Yapışkan fibrin pıhtısı ile etkileşim olmadığından emin olun. </li><li> Fibrin pıhtıları konfokal görüntüleme önce 2 saat 21-23 ° C&#39;de polimerize izin verin. </li><li> 488 nm Argon lazer ile 40X / 1.30 Yağ M27 lens ile bir konfokal mikroskop kullanılarak pıhtıları konfokal mikroskopi görüntüleri çekmek. </li></ol> Orak Hücre Hastalarda gelen eritrositlerde İçeren Fibrin pıhtısı 3. Konfokal Mikroskopi Değerlendirme <ol><li> 37 ° C &#39;de, insan fibrinojen ve floresan etiketli insan fibrinojen konjugatı defrost. </li><li> Pipet 0.5 ml aşağıdaki mikrosantrifüj tüp mezun oldu. <ol><li> Sağlıklı normal fibrin pıhtısı için, 5 mg / ml&#39;lik bir konsantrasyonda 50 mM Tris / 100 mM NaCI tampon maddesi içinde% 10 floresan etiketli insan fibrinojen konjügatı ile insan fibrinojen karıştırın. </li><li> SCD eritrositlerde ihtiva eden pıhtılar için, fibrinojen sonuçtaki konsantrasyonunun 10 mg / ml olan bir 50 mM Tris / 100 mM NaCI gibi insan fibrinojen ve% 10 floresan etiketli insan fibrinojen konjugatı karıştırın. </li></ol></li><li> Bir hücre-etiketleme çözümü kullanılarak izole eritrositlerde (normal ve RBC izolasyon protokolü elde orak hücre hastalardan olanlar ikisi) Etiket. <ol><li> Seçilen herhangi bir serum içermeyen kültür ortamı içinde ml başına 1 x 10 6 yoğunluğunda süspanse. </li><li> Hücre etiketleme çözeltisine hücre süspansiyonu 5 ul / ml ilave edilir. Nazik pipetleme ile iyice karıştırın yavaşça. </li><li&gt; 37 ° C &#39;de 20 dakika boyunca inkübe edin. </li><li> 37 ° C&#39;de 5 dakika boyunca 1500 rpm&#39;de etiketli süspansiyon tüpleri santrifüjleyin. </li><li> 37 ° C orta hücreleri yeniden askıya hafifçe Süpernatantı ve. </li><li> Yıkama işlemi (3.3.4 ve 3.3.5) iki kez daha tekrarlayın. </li></ol></li><li> 5 mM CaCl2 tampon içinde 80 U / ml FXIIIa seyreltin. </li><li> 5 mM CaCl2 tamponu içinde 4 U / ml trombin seyreltin. <ol><li> Normal pıhtılar için, fibrinojen, FXIIIa ve trombin nihai konsantrasyonları sırası ile 50 ul&#39;lik bir hacimde, 2.5 mg / ml, 20 U / ml ve 1 U / ml olduğundan emin olun. </li><li> SCD eritrositlerde ihtiva eden pıhtılar için, fibrinojen, FXIIIa ve trombin nihai konsantrasyonları sırası ile 50 ul&#39;lik bir hacimde 5 mg / ml, 20 U / ml ve 1 U / ml olduğundan emin olun. </li></ol></li><li> Pipet fibrin çözeltisi içine etiketli eritrosit 10 ul. Pipet mikroskop camı üzerine eritrosit ile fibrin 30 ul (gl için hazırlık slayt bakınycated pıhtıları). </li><li> Fibrin konfokal görüntüleme önce 2 saat 21-23 ° C&#39;de polimerize izin verin. </li><li> Konfokal mikroskop kullanılarak pıhtıları konfokal görüntüleri çekmek ve floresan etiketli fibrin ve eritrositlerde heyecanlandırmak için 488 nm ve 633 nm dalga boylarında ile uyarım lazerleri kullanıyoruz. </li></ol> Glike pıhtı Yapılarında Fibrinolizis Oranları 4. Konfokal Mikroskopi Değerlendirme <ol><li> Glike pıhtıları içinde fibrinoliz oranı değerlendirmesi için glike pıhtıları (Protokol Adım 2) konfokal mikroskopi protokolünü tekrarlayın. </li><li> Net yapıştırıcı ile odasının uçlarını mühür yok. Fibrin pıhtısı kaybını önlemek için 250 ml suyla ihtiva eden kapalı bir muhafaza içinde 21-23 ° C de 1.5 saat süre ile polimerize olmaya bırakın. </li><li> Polimerizasyon sonra, konfokal mikroskop kullanılarak pıhtı görüntüleri elde edebilir. </li><li> Pipet bölmesinin açık ucuna plazmin ve kılcal hareket aracılığı ile pıhtı ile plazmin dağıtılır. <ol><li> IçinNormal konsantrasyonlar, pipet odasının açıklığı içine plazmin, 200 ug / ml&#39;de 25 ul normal glikozile fibrinoliz deneyleri. </li><li> Düşük konsantrasyonlarda pipet odasının açıklığı içine 50 ug / ml&#39;de 25 ul ile glikolize pıhtı için. </li></ol></li><li> Pıhtı yokedici plazminin gerçek zamanlı video görüntüleri yakalamak için konfokal mikroskop yazılımındaki zaman serisi özelliğini (bkz Şekil 1) kullanın. <ol><li> Satın alma modunu tıklayın ve ardından &quot;Zaman Serisi.&quot; Döngü sayısını ve kayıt süresi aralığını seçin. </li></ol></li></ol> 5. Fibrinolizis Fiyatlarını Hesaplama <ol><li> Pıhtı sabit alanı çözmek için gerekli geçen süreyi bir alanı (2,500 mikron 2) ayarlayarak ve hesaplayarak parçalama hızını belirleyin. Aşağıdaki denklem (Şekil 2) kullanılarak parçalama hızını hesaplayın: <img alt="Denklem 1" fo:content-width="2in" src = &quot;/ files / ftp_upload / 52.019 / 52019eq1.jpg&quot; /&gt; </li></ol> Fibrinolizis Oranlarının 6. İstatistiksel Analiz <ol><li> İstatistiksel analiz yazılımı kullanılarak lizis verileri analiz. Bir tek-yönlü ANOVA ve hoc Tukey-Kramer testi 44,45 bir yazı gerçekleştirin. </li></ol>

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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fibrinogen+and+fibrin.">Fibrinogen and fibrin.</a> Advances in protein chemistry. 70, 247-299 (2005).</li><li>Collet, 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=Influence+of+Fibrin+Network+Conformation+and+Fibrin+Fiber+Diameter+on+Fibrinolysis+Speed+Dynamic+and+Structural+Approaches+by+Confocal+Microscopy.">Influence of Fibrin Network Conformation and Fibrin Fiber Diameter on Fibrinolysis Speed Dynamic and Structural Approaches by Confocal Microscopy.</a> Arteriosclerosis, thrombosis, and vascular biology. 20, 1354-1361 (2000).</li><li>J&#246;rneskog, G., 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=Altered+properties+of+the+fibrin+gel+structure+in+patients+with+IDDM.">Altered properties of the fibrin gel structure in patients with IDDM.</a> Diabetologia. 39, 1519-1523 (1996).</li><li>Svensson, 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=Acetylation+and+glycation+of+fibrinogen+in+vitro+occur+at+specific+lysine+residues+in+a+concentration+dependent+manner:+A+mass+spectrometric+and+isotope+labeling+study.">Acetylation and glycation of fibrinogen in vitro occur at specific lysine residues in a concentration dependent manner: A mass spectrometric and isotope labeling study.</a> Biochemical and Biophysical Research Communications. 421, 335-342 (2012).</li><li>Pieters, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycation+of+fibrinogen+in+uncontrolled+diabetic+patients+and+the+effects+of+glycaemic+control+on+fibrinogen+glycation.">Glycation of fibrinogen in uncontrolled diabetic patients and the effects of glycaemic control on fibrinogen glycation.</a> Thrombosis Research. 120, 439-446 (2007).</li><li>Baradet, T. C., Haselgrove, J. C., Weisel, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+reconstruction+of+fibrin+clot+networks+from+stereoscopic+intermediate+voltage+electron+microscope+images+and+analysis+of+branching.">Three-dimensional reconstruction of fibrin clot networks from stereoscopic intermediate voltage electron microscope images and analysis of branching.</a> Biophysical journal. 68, 1551-1560 (1995).</li><li>Campbell, R. A., Overmyer, K. A., Selzman, C. H., Sheridan, B. C., Wolberg, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contributions+of+extravascular+and+intravascular+cells+to+fibrin+network+formation,+structure,+and+stability.">Contributions of extravascular and intravascular cells to fibrin network formation, structure, and stability.</a> Blood. 114, 4886-4896 (2009).</li><li>Curtis, D. 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=A+study+of+microstructural+templating+in+fibrin-thrombin+gel+networks+by+spectral+and+viscoelastic+analysis.">A study of microstructural templating in fibrin-thrombin gel networks by spectral and viscoelastic analysis.</a> Soft Matter. 9, 4883-4889 (2013).</li><li>Kim, E., 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=Correlation+between+fibrin+network+structure+and+mechanical+properties:+an+experimental+and+computational+analysis.">Correlation between fibrin network structure and mechanical properties: an experimental and computational analysis.</a> Soft Matter. 7, 4983-4992 (2011).</li><li>Keegan, P. M., Surapaneni, S., Platt, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sickle+cell+disease+activates+peripheral+blood+mononuclear+cells+to+induce+cathepsins+k+and+v+activity+in+endothelial+cells.">Sickle cell disease activates peripheral blood mononuclear cells to induce cathepsins k and v activity in endothelial cells.</a> Anemia. 2012, 201781 (2012).</li><li>Allison, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+sickle-cell+haemoglobin.">Properties of sickle-cell haemoglobin.</a> Biochemical Journal. 65, 212 (1957).</li><li>Kuehl, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Design of experiments : statistical principles of research design and analysis. , (2000).</li><li>Lindman, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Analysis of variance in experimental designs. , (1992).</li><li>Pan, W., Galkin, O., Filobelo, L., Nagel, R. L., Vekilov, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metastable+Mesoscopic+Clusters+in+Solutions+of+Sickle-Cell+Hemoglobin.">Metastable Mesoscopic Clusters in Solutions of Sickle-Cell Hemoglobin.</a> Biophysical Journal. 92, 267-277 (2007).</li><li>Wootton, D. M., Popel, A. S., Alevriadou, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+and+theoretical+study+on+the+dissolution+of+mural+fibrin+clots+by+tissue-type+plasminogen+activator.">An experimental and theoretical study on the dissolution of mural fibrin clots by tissue-type plasminogen activator.</a> Biotechnology and bioengineering. 77, 405-419 (2002).</li><li>Sazonova, I. Y., 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=127+Mathematic+model+of+fibrin+clot+lysis+by+plasmin.">127 Mathematic model of fibrin clot lysis by plasmin.</a> Fibrinolysis and Proteolysis. 12, 45 (1998).</li></ol>

The authors have nothing to disclose.

Hastalık durumlarında, pıhtılaşma mekanizmalarının yapısı anlamlı veriler elde etmek için, bu koşullar altında, protein ve hücre etkilerini belirlemek için pıhtılaşma faktörleri izole etmek için önemlidir. Bu protokol in vitro diyabetik ve OHA devletler fibrin pıhtı yapısını araştıran amacıyla geliştirilmiştir. Bu değiştirilmiş koşullar hiperkoagülasyon, Aterotrombozu ve KVH neden beri hastalık durumlarında fibrin oluşumu ve fibrinoliz yer alan...

Bir kan geminin endotel astar yaralanması hemostatik yanıt, ya da bir kan pıhtısı oluşumu ile tamir edilir. Kan hücre dışı matris içine nüfuz zaman, doku faktörleri koagülasyon kaskad başlatılmasını kolaylaştırmak, kan akışında trombositleri aktive. Bu iyileşme sürecinin önemli mekanik parça son derece esnek olan fibrin liflerden oluşan fibrin matrisi, ve büyük kuvvetler 1-4 muhafaza eder. Birçok araştırmacı yoğun geçmiş yıllarda 5-13 fibrin oluşumu yapısı ve fonksiyonu inceledik. Diabetes mellitus ve orak hücre olarak hastalıkları olan hastalar gibi miyokard enfarktüsü, iskemik kalp hastalığı gibi trombotik komplikasyonların gelişme riski, ve 14-19 okşar. 2 milyondan fazla insan yeni Amerika Birleşik Devletleri&#39;nde her yıl diyabet tanısı konur. Tip I nerede,: diyabet iki türü vardırvücut insülin dirençli hale insülin ve Tip II, yeterli miktarda üretemez. Diyabetik hastalarda, kardiyovasküler hastalık (CVD) hastalığı 20,21 ile ilişkili morbidite ve mortalitenin% 80 nedenidir. Orak hücre hastalığı (SCD) Amerika Birleşik Devletleri 22 100.000&#39;den fazla insanı etkileyen bir genetik kan hastalığıdır. OHA zor hücreler, kan damar 23 geçmesi için yapım, hilal şeklinde olmak üzere, kırmızı kan hücrelerinin neden olan bir nokta mutasyon hastalıktır. Bu hastalık durumlarının her iki gövde aterosklerotik durumları geliştirme ihtimalini artırır. Bunun nedenlerinden biri, hastalıklı devletlerin 14,24-26 değişmiş fibrin yapısı ve fonksiyonu bir sonucudur. Her iki diyabet ve orak hücre hastalığında, hiperkoagülasyon ve Aterotrombozu ve kardiyovasküler hastalık neden olur hipofibrinolizin aktivitesi (C varVD) sağlıklı hastalarda 17,27,28 ile karşılaştırıldığında. Bu hipofibrinolizin aterosklerozun ilerlemesini teşvik eder ve erken koroner arter hastalığı 29 olan hastalar için tekrarlayan iskemik olayları doğurur bilinmektedir. Mevcut yazıda, bu özel ortamda fibrin fiziksel özelliklerinin rolü araştırıldı. Olmayan hastalıklı hastalarda fibrin pıhtısı yapıları, ince lifler, daha büyük gözenekleri ve daha az yoğun, genellikle 14,24 oluşmaktadır. Sağlıklı hastalarda artan bir geçirgenliğe ve daha az yoğun bir fibrin pıhtısı fibrinoliz 16 kolaylaştırmak için tespit edilmiştir. Diyabetik ve orak hücre hastalığı gibi hyperthrombotic koşullarda sağlıklı hastalarda 30-33 2.5 mg / ml, normal seviyelerine artış fibrinojen neden fibrinojen üretiminde bir artış vardır. Sağlıklı, sigara dia göre diyabetik hastalarda oluşan fibrin pıhtılar daha fazla dal noktalarını, ve daha yoğun, daha sert ve daha az gözenekli olduğu tespit edilmiştirdiyabetik hastalarda 14,24,33-35. değiştirilmiş fibril bağları pıhtı oluşumunda rol oynayan proteinlerde meydana gelen glikozilasyon mekanizmasının bir sonucudur. Glukoz molekülleri, uygun bir şekilde çapraz bağlama glutamin ve lisin tortuları 33,36,37 insan faktör XIIIa (FXIIIa) inhibe fibrinojen molekülü, lizin artıklarının bağlanan zaman Enzimatik olmayan (geri dönüşümsüz) glikasyonu oluşur. Fibrin ağlarının yapısal analiz son zamanlarda yoğun çalışılmıştır. Özellikle, araştırmacılar elektron mikroskobu ve intravasküler (endotel) hücreleri ve ekstravasküler (fibroblastlar ve düz kas) hem hücreler fibrin yapıları 40 analiz fibrin yapısı 39, kullanılan viskoelastik ve spektral analiz nasıl etkilediğini araştırdık fibrin ağları 38, 3D rekonstrüksiyon ve kullanmıştır Fibrin yapısı ve mekanik özellikleri arasında gelişmiş korelasyon deneysel kullanarak ve hesaplamalı yaklaşımlar 41 </sup&gt;. Bu çalışmanın odak simüle diyabetik ve orak hücreli tromboz koşulları altında pıhtı yapıları formüle etmek ve hastalıklı devletlerin pıhtı yapısının incelenmesi ve işlevi için konfokal mikroskopi kullanımı idi. Fibrin pıhtılar insan fibrinojen, insan trombini ve FXIIIa oluşmuş. pıhtılar plazmin kullanılarak lize edilmiştir. Diyabetik koşulların simüle edilmesi için, fibrinojen artan konsantrasyonu, in vitro olarak fibrinojen glikasyonunu uyarılması için glikoz çözeltisi içinde kuluçkalanmıştır. Orak hücre hastalığı pıhtılaşma koşulları simüle etmek için, artmış fibrinojen konsantrasyonları grubumuza 42 tarafından daha önce yapıldığı gibi hastalardan toplanan orak hücre hematokrit ile karıştırıldı. Bu yöntemler, yapı ve hastalıklı koşullar altında, fibrin pıhtısı oluşumu ve fibrinolizde rol alan ve fonksiyonları, yanı sıra CVD neden mekanizmaları incelemek için kullanılmıştır. Bu hastalıklar hakkında güncel bilgilere dayanarak, glike fibrin pıhtı yapıları daha az a ile yoğun olduğunund küçük gözenekler. Orak hücre hastalarında (eritrositler) kırmızı kan hücreleri ile fibrin pıhtıları da yoğun ve eritrosit agregasyonunu ve aglomere fibrin kümeleri görüntülenir. Bu, daha önce 43 tespit edilmiştir köklü bir olgudur. Ayrıca fibrinoliz oranı ile sağlıklı, normal bir fibrin kıyasla azalmış plazmin olmadan glike fibrin pıhtıları anlamlı olarak düşük olacağı varsayılmıştır. Sonuçlar, glikolize fibrin pıhtılaşması için önemli ölçüde farklı lisis oranı sonuçları yalnızca sınırlı plazmin konsantrasyonu durumunda görüldüğünü göstermiştir. Hücreler ve proteinler pıhtılaşma faaliyetinin gerçek zamanlı video yakalama sağlayan kendi ana devlet kalır, çünkü konfokal mikroskopi kullanılarak bu deneysel tekniği diğer görüntüleme yöntemlerine göre önemli avantajlar sunmaktadır. Sentetik indükleyici pıhtılaşma Bu yöntem, hasta örnekleri elde etmek ve bireysel filtreleyerek daha da ucuz ve daha fazla zaman verimliproteinler ve enzimler. Örnekler arasında değişkenlik plazma diğer proteinler sonucu olmadığını Bundan başka, pıhtı sentezlenmesi için ayrılmış proteinler ve enzimler kullanarak, pıhtılar standardize edildi.

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			<td height="21" style="height:21px;width:315px;">Name of Material/ Equipment</td>
			<td style="width:287px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:315px;">PBS</td>
			<td style="width:287px;">Life Technologies</td>
			<td style="width:119px;">10010031</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:315px;">Ficoll-Paque (hydrophilic polysaccharide)</td>
			<td style="width:287px;">GE Healthcare</td>
			<td style="width:119px;">45-001-749</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 ml heparinized vacutainer tubes</td>
			<td style="width:287px;">BD Biosciences</td>
			<td style="width:119px;">366643</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:315px;">Human Fibrinogen</td>
			<td style="width:287px;">Enzyme Research Laboratories</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Alexa Fluor 488 human fibrinogen conjugate</td>
			<td style="width:287px;">Molecular Probes</td>
			<td style="width:119px;">F13191</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.5 mL graduated microcentrifuge tube</td>
			<td style="width:287px;">Fisher Scientific</td>
			<td style="width:119px;">05-408-120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:315px;">Glucose powder</td>
			<td style="width:287px;">Life Technologies</td>
			<td style="width:119px;">15023-02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:315px;">FXIIIa</td>
			<td style="width:287px;">Enzyme Research Laboratories</td>
			<td style="width:119px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">VIS Confocal Microscope</td>
			<td style="width:287px;">Zeiss LSM 510</td>
			<td style="width:119px;">LSM 510</td>
			<td></td>
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			<td height="21" style="height:21px;width:315px;">50 mM Tris</td>
			<td style="width:287px;">Lonza</td>
			<td style="width:119px;">S50-642</td>
			<td></td>
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		<tr height="25">
			<td height="25" style="height:25px;width:315px;">Calcium Chloride (CaCl2)</td>
			<td style="width:287px;">Sigma Aldrich</td>
			<td style="width:119px;">449709-10G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vybrant DiD cell-labeling solution</td>
			<td style="width:287px;">Life Technologies</td>
			<td style="width:119px;">L7781</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:315px;">Plasmin</td>
			<td style="width:287px;">Enzyme Research Laboratories</td>
			<td style="width:119px;">N/A</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:315px;">Sodium Chloride (5 M NaCl)</td>
			<td style="width:287px;">Life Technologies</td>
			<td style="width:119px;">AM9759</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:315px;">Statistical Modeling Software</td>
			<td>IBM</td>
			<td style="width:119px;">SPSS Statistics 22</td>
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experimental and imaging techniques for examining fibrin clot structures in normal and diseased states

Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on fibrin in the past years to include the investigation of synthesis, structure-function, and lysis of clots. However, there is still much unknown about the morphological and structural features of clots that ensue from patients with disease. In this research study, experimental techniques are presented that allow for the examination of morphological differences of abnormal clot structures due to diseased states such as diabetes and sickle cell anemia. Our study focuses on the preparation and evaluation of fibrin clots in order to assess morphological differences using various experimental assays and confocal microscopy. In addition, a method is also described that allows for continuous, real-time calculation of lysis rates in fibrin clots. The techniques described herein are important for researchers and clinicians seeking to elucidate comorbid thrombotic pathologies such as myocardial infarctions, ischemic heart disease, and strokes in patients with diabetes or sickle cell disease.

Glike Fibrin pıhtı Yapıların Konfokal Mikroskopi Analizi Normal ve glikolize pıhtı konfokal mikroskobu görüntüleri. Şekil 3&#39;te sunulmuştur normal ve glikolize pıhtı mikroskopisi analizi Glike pıhtılar daha yoğun olan ve pıhtı polimerizasyon sırasında FXIIIa ilavesi olmadan hem normal pıhtıları daha küçük gözeneklere sahip olduğunu ortaya koymaktadır. Şekil 3A ve 3B, daha yüksek bir fibrin konsantrasyonu <st...

Watch this Scientific Journal Video about Experimental and Imaging Techniques for Examining Fibrin Clot Structures in Normal and Diseased States at JoVE.com

Experimental and Imaging Techniques for Examining Fibrin Clot Structures in Normal and Diseased States

In this manuscript, experimental techniques, including blood preparation, confocal microscopy, and lysis rate analysis, to examine the morphological differences between normal and abnormal clot structures due to diseased states are presented.

Normal ve Hastalıklı Devletleri&#39;nde Fibrin pıhtısı Yapıları incelenmesi için deneysel ve Görüntüleme Teknikleri

Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on ...

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11.3K Görüntüleme.  Georgia Institute of Technology & Emory University School of Medicine. In this manuscript, experimental techniques, including blood preparation, confocal microscopy, and lysis rate analysis, to examine the morphological differences between normal and abnormal clot structures due to diseased states are presented.

Video: Experimental and Imaging Techniques for Examining Fibrin Clot Structures in Normal and Diseased States

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular cartilage and subchondral bone in recent years led to increased attention to restoration of the entire osteochondral unit. In comparison to chondral lesions the regeneration of osteochondral defects is much more complex and a far greater surgical and therapeutic challenge. The damaged tissue does not only include the superficial cartilage layer but also the subchondral bone. For deep, osteochondral damage, as it occurs for example with osteochondrosis dissecans, the full thickness of the defect needs to be replaced to restore the joint surface 1. Eligible therapeutic procedures have to consider these two different tissues with their different intrinsic healing potential 2. In the last decades, several surgical treatment options have emerged and have already been clinically established 3-6.
Autologous or allogeneic osteochondral transplants consist of articular cartilage and subchondral bone and allow the replacement of the entire osteochondral unit. The defects are filled with cylindrical osteochondral grafts that aim to provide a congruent hyaline cartilage covered surface 3,7,8. Disadvantages are the limited amount of available grafts, donor site morbidity (for autologous transplants) and the incongruence of the surface; thereby the application of this method is especially limited for large defects.
New approaches in the field of tissue engineering opened up promising possibilities for regenerative osteochondral therapy. The implantation of autologous chondrocytes marked the first cell based biological approach for the treatment of full-thickness cartilage lesions and is now worldwide established with good clinical results even 10 to 20 years after implantation 9,10. However, to date, this technique is not suitable for the treatment of all types of lesions such as deep defects involving the subchondral bone 11.
The sandwich-technique combines bone grafting with current approaches in Tissue Engineering 5,6. This combination seems to be able to overcome the limitations seen in osteochondral grafts alone. After autologous bone grafting to the subchondral defect area, a membrane seeded with autologous chondrocytes is sutured above and facilitates to match the topology of the graft with the injured site. Of course, the previous bone reconstruction needs additional surgical time and often even an additional surgery. Moreover, to date, long-term data is missing 12.
Tissue Engineering without additional bone grafting aims to restore the complex structure and properties of native articular cartilage by chondrogenic and osteogenic potential of the transplanted cells. However, again, it is usually only the cartilage tissue that is more or less regenerated. Additional osteochondral damage needs a specific further treatment. In order to achieve a regeneration of the multilayered structure of osteochondral defects, three-dimensional tissue engineered products seeded with autologous/allogeneic cells might provide a good regeneration capacity 11.
Beside autologous chondrocytes, mesenchymal stem cells (MSC) seem to be an attractive alternative for the development of a full-thickness cartilage tissue. In numerous preclinical in vitro and in vivo studies, mesenchymal stem cells have displayed excellent tissue regeneration potential 13,14. The important advantage of mesenchymal stem cells especially for the treatment of osteochondral defects is that they have the capacity to differentiate in osteocytes as well as chondrocytes. Therefore, they potentially allow a multilayered regeneration of the defect.
In recent years, several scaffolds with osteochondral regenerative potential have therefore been developed and evaluated with promising preliminary results 1,15-18. Furthermore, fibrin glue as a cell carrier became one of the preferred techniques in experimental cartilage repair and has already successfully been used in several animal studies 19-21 and even first human trials 22.
The following protocol will demonstrate an experimental technique for isolating mesenchymal stem cells from a rabbit's bone marrow, for subsequent proliferation in cell culture and for preparing a standardized in vitro-model for fibrin-cell-clots. Finally, a technique for the implantation of pre-established fibrin-cell-clots into artificial osteochondral defects of the rabbit's knee joint will be described.

Treatment of Osteochondral Defects in the Rabbit&#039;s Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

An experimental technique for the treatment of osteochondral defects in the rabbit's knee joint is described. The implantation of allogeneic mesenchymal stem cells into osteochondral defects provides a promising development in the field of tissue engineering. The preparation of fibrin-cell-clots in vitro offers a standardized method for implantation.

Fibrin pıhtısı içinde allojenik Mezenkimal Kök Hücre implantasyonu ile Tavşan Diz Eklemi içinde Osteokondral Kusur tedavisi

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

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A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

Yeni Bir Yüksek çözünürlüklü<em&gt; In vivo</emTümör Büyüme ve Tedavi için İntrakranial Yapıların Dinamik Tepki Eğitim&gt; Görüntüleme Tekniği

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

Implanting Glass Spinal Cord Windows in Adult Mice with Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as multiple sclerosis. Here we present a low-cost and reproducible glass window implantation protocol that is suitable for intravital microscopy and studying the dynamics of spinal cord cytoarchitecture with subcellular resolution in live adult mice with EAE. Briefly, we surgically expose the vertebrae T12-L2 and construct a chamber around the exposed vertebrae using a combination of cyanoacrylate and dental cement. A laminectomy is performed from T13 to L1, and a thin layer of transparent silicone elastomer is applied to the dorsal surface of the exposed spinal cord. A modified glass cover slip is implanted over the exposed spinal cord taking care that the glass does not directly contact the spinal cord. To reduce the infiltration of inflammatory cells between the window and spinal cord, anti-inflammatory treatment is administered every 2 days (as recommended by ethics committee) for the first 10 days after implantation. EAE is induced only 2-3 weeks after the cessation of anti-inflammatory treatment.
Using this approach we successfully induced EAE in 87% of animals with implanted windows and, using Thy1-CFP-23 mice (blue axons in dorsal spinal cord), quantified axonal loss throughout EAE progression. Taken together, this protocol may be useful for studying the recruitment of various cell populations as well as their interaction dynamics, with subcellular resolution and for extended periods of time. This intravital imaging modality represents a valuable tool for developing therapeutic strategies to treat autoimmune demyelinating diseases such as EAE.

We describe a method for implanting and maintaining glass windows over the exposed spinal cords of adult mice for studying experimental autoimmune encephalomyelitis. These windows provide chronic optical access to the spinal cord for monitoring cell dynamics at the subcellular level in live animals using two-photon microscopy.

Deneysel Otoimmün Encephalomyelitis Erişkin Farelerde Cam Omurilik Windows&#39;u implante

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as ...

A Pipeline for 3D Multimodality Image Integration and Computer-assisted Planning in Epilepsy Surgery

Epilepsy surgery is challenging and the use of 3D multimodality image integration (3DMMI) to aid presurgical planning is well-established. Multimodality image integration can be technically demanding, and is underutilised in clinical practice. We have developed a single software platform for image integration, 3D visualization and surgical planning. Here, our pipeline is described in step-by-step fashion, starting with image acquisition, proceeding through image co-registration, manual segmentation, brain and vessel extraction, 3D visualization and manual planning of stereoEEG (SEEG) implantations. With dissemination of the software this pipeline can be reproduced in other centres, allowing other groups to benefit from 3DMMI. We also describe the use of an automated, multi-trajectory planner to generate stereoEEG implantation plans. Preliminary studies suggest this is a rapid, safe and efficacious adjunct for planning SEEG implantations. Finally, a simple solution for the export of plans and models to commercial neuronavigation systems for implementation of plans in the operating theater is described. This software is a valuable tool that can support clinical decision making throughout the epilepsy surgery pathway.

We describe the steps to use our custom designed software for image integration, visualization and planning in epilepsy surgery.

Epilepsi Cerrahisi 3D Multimodalite Görüntü Entegrasyonu ve Bilgisayar destekli Planlama Bir Boru Hattı

Epilepsy surgery is challenging and the use of 3D multimodality image integration (3DMMI) to aid presurgical planning is well-established. Multimodality ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

Normal Farelerde% 70 hepatektomi sonrası Vasküler ve Parankimal Rejenerasyon görselleştirme

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming more commonly used in trauma research. They offer a large number of mutant strains and antibodies for the analysis of the molecular mechanisms behind the highly differentiated process of bone healing. To control the biomechanical environment, standardized and well-characterized osteosynthesis techniques are mandatory in mice. Here, we report on the design and use of an intramedullary nail to stabilize open femur osteotomies in mice. The nail, made of medical-grade stainless steel, provides high axial and rotational stiffness. The implant further allows the creation of defined, constant osteotomy gap sizes from 0.00 mm to 2.00 mm. Intramedullary locking nail stabilization of femur osteotomies with gap sizes of 0.00 mm and 0.25 mm result in adequate bone healing through endochondral and intramembranous ossification. Stabilization of femur osteotomies with a gap size of 2.00 mm results in atrophic non-union. Thus, the intramedullary locking nail can be used in healing and non-healing models. A further advantage of the use of the nail compared to other open bone healing models is the possibility to adequately fix bone substitutes and scaffolds in order to study the process of osseous integration. A disadvantage of the use of the intramedullary nail is the more invasive surgical procedure, inherent to all open procedures compared to closed models. A further disadvantage may be the induction of some damage to the intramedullary cavity, inherent to all intramedullary stabilization techniques compared to extramedullary stabilization procedures.

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

Femur Osteotomiler standardize Sabitleme için bir intramedüller Kilitleme Nail Fare Normal ve Arızalı Kemik Şifa Analiz

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming ...

In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging

Cerebral malaria is a sign of severe malarial disease and is often a harbinger of death. While aggressive management can be life-saving, the detection of cerebral malaria can be difficult. We present an experimental mouse model of cerebral malaria that shares multiple features of the human disease, including edema and microvascular pathology. Using magnetic resonance imaging (MRI), we can detect and track the blood-brain barrier disruption, edema development, and subsequent brain swelling. We describe multiple MRI techniques that can visualize these pertinent pathological changes. Thus, we show that MRI represents a valuable tool to visualize and track pathological changes, such as edema, brain swelling, and microvascular pathology, in vivo.

In Vivo Tracking of Edema Development and Microvascular Pathology in a Model of Experimental Cerebral Malaria Using Magnetic Resonance Imaging

We describe a mouse model of experimental cerebral malaria and show how inflammatory and microvascular pathology can be tracked in vivo using magnetic resonance imaging.

Manyetik Rezonans Görüntüleme Kullanarak Deneysel Serebral Sıtma Bir Modelinde Ödem Gelişimi ve Mikrovasküler Patolojinin İn Vivo Takibi

Cerebral malaria is a sign of severe malarial disease and is often a harbinger of death. While aggressive management can be life-saving, the detection of ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Bir Fulminan Miyokardit Modelinde Işık Yeri Mikroskopisi İle Lökosit İnfiltrasyonunun Nicel Görselleştirilmesi

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

In Vivo Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Meni&#232;re's Disease Ears and Normal Hearing Ears

Analysis of neural structures in Meni&#232;re's Disease (MD) is of importance, since a loss of such structures has previously been proposed for this patient group but has yet to be confirmed. This protocol describes a method of in vivo evaluation of neural changes especially well suitable for cranial nerve analysis using magnetic resonance imaging (MRI). MD-patients and normal hearing persons were examined in a 3-T MR-scanner using a scan protocol including strongly T2-weighted 3D gradient-echo-sequence (3D-CISS). In the patient group, MD was additionally confirmed using MRI-based assessment of endolymphatic hydrops. Morphometric analysis was performed using a freeware DICOM viewer. Evaluation of cranial nerves included measurements of cross-sectional areas (CSAs) of the nerves at different levels as well as orthogonal diametric measurements.

&lt;em&gt;In Vivo&lt;/em&gt; Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Meni&amp;#232;re&#039;s Disease Ears and Normal Hearing Ears

To evaluate morphological changes of cranial nerves such as loss of neural structures or swelling of cranial nerves in Meni&#232;re&#39;s Disease (MD) or in healthy persons in vivo, a protocol of evaluation has been developed using magnetic resonance imaging (MRI). Additional MRI-based confirmation of MD was performed.

Vivo Menière'nın hastalığı kulakları ve Normal işitme kulak manyetik rezonans görüntüleme kullanarak insan kranial sinirler xarakteristikaları analizi

Analysis of neural structures in Meni&#232;re's Disease (MD) is of importance, since a loss of such structures has previously been proposed for this ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The appearance of a mass on certain magnetic resonance (MR) sequences can help differentiate a thrombus from competing diagnoses such as a tumor. T1 and T2 signal characteristics of a thrombus are related to the evolution of hemoglobin properties. A thrombus typically does not enhance following contrast administration, which also helps differentiation from a tumor. We also highlight the emerging role of T1 mapping in the evaluation of a thrombus, which can add another level of support in diagnosis. Prior to any CMR exam, patient screening and interviews are critical to ensure safety and to optimize patient comfort. Effective communication during the exam between the technologist and the patient promotes proper breath holding technique and higher quality images. Volumetric post processing and structured reporting are helpful to ensure that the radiologist answers the ordering services&#39; question and communicates these results effectively. Optimal pre-MR safety evaluation, CMR exam execution, and post exam processing and reporting allow for delivery of high quality radiological service in the evaluation of a suspected cardiac thrombus.

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

Şüpheli kardiyak Trombülerin değerlendirilmesi için kardiyak manyetik rezonans: konvansiyonel ve gelişmekte olan teknikler

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...