Biz Drs kabul ediyorsunuz. Camillo Ricordi, Antonello Pileggi, R. Damaris Molano, Stephan Speier ve verimli tartışmalara Daniel Nyqvist. Ayrıca video kayıt ile yardım Eleut Hernandez ve Diego Espinosa-Heidmann teknik yardım için, ve Mike Valdes ve Margaret Formoso ederim. Byron Maldonado, düzenlenebilir kaydedilir ve son video üretti. Araştırma destek Diyabet Araştırma Enstitüsü Vakfı (tarafından sağlanmıştır <a href="http://www.DiabetesResearch.org" target="_blank">www.DiabetesResearch.org</a> ), NIH / NIDDK / NIAID (MHA için F32DK083226; AC NIH RO3DK075487; PO.B. için U01DK089538). PO.B Ek araştırma desteği Karolinska Enstitüsünde, İsveç Araştırma Konseyi, İsveç Diyabet Vakfı, Aile Erling-Persson Vakfı, Aile Knut ve Alice Wallenberg Vakfı, VIBRANT Skandia Sigorta Şirketi Ltd, fonlar aracılığıyla sağlanan ( Karolinska Enstitüsü de Diyabet FP7-228933-2), Stratejik Araştırma Programıitutet, Novo Nordisk Vakfı ve Çekek von Kantzow Vakfı.

Aşağıdaki yordam 2 adım stereoskop altında yapılır, ilk adım kanül içine adacıklar yükleme içerir ve ikinci adım ACE fiili naklidir. Hayvanlar üzerinde yapılan tüm işlemler Miami Üniversitesinin kurumsal hayvan bakımı ve kullanımı komitesi (IACUC) tarafından onaylanmıştır. 1. Transplantasyon için Kanül Yükleme adacıkları <ol><li> Daralma çevrelerinde çanak iplik Merkezi kültürü çanak adacıklar. </li><li> &quot;Rezervuar&quot; dan kanül ayırın ve temiz bir yüzey üzerinde kanül ve bağlantı hortumu yerleştirin. Rezervuar filtresi (Şekil 1a) olmadan 300 ul tek kullanımlık plastik pipet üzerinden yapılabilir. </li><li> Rezervuar içine aspirasyon olduğunda adacık sürekli akış sağlamak için rezervuarın hava kabarcıklarının dışarı yıkayın. Rezervuar Flushing ayak pedalı kullanarak eller serbest motorlu şırınga sürücüsü ileri sürerek yapılır(Şekil 1B, C). Bu da rezervuar (örneğin tuz, PBS veya kültür ortamı gibi steril solüsyonu ile önceden yüklenmiş olarak) içine adacık aspirasyon izin şırınga boşluk yapacaktır. </li><li> Yavaşça rezervuar içine adacık istenilen miktarda aspire. Onlar rezervuar girin ve dibe doğru birlikte olarak kalacaktır adacıkları girdap eğiliminde olacaktır. Aspirasyon ayak pedalı kullanarak geriye motorlu şırınga sürücü sürüş yapılır. </li><li> Bağlayan tüp yoluyla rezervuar kanül takın. </li><li> Kültür çanak geri kanül ucu yerleştirin ve kanül içine daha sonra boru içine rezervuarın adacıkları dışarı floş. Eğer hafifçe (çekme) boru (Şekil 1d) &quot;Flicking&quot; ile tüp / kanül geri dolgu olarak adacık birlikte kalmasını sağlayın. Kanül dışarı atılır öncesinde adacıklar tüm hava kabarcıkları önce veya sonra durdurun. Emin değilseniz, adacıklar kalan hava gibi kanül geri girmek olarak durdurmaköncesinde adacıklar baloncuklar ACE adacık reflü (geri akış) üzerinden önlemeye yardımcı olabilir. Gecede dağılacaktır. </li><li> Bu aşamada, ACE (sonraki adımlara bakın lütfen) içine adacıklar enjekte hazırız. </li></ol> 2. Göz Ön Kamara içine Islet Nakli <ol><li> Stereoskop altında sıcak bir pad üzerinde anestezi fare yerleştirin. </li><li> Oksijen / izofluran anestezisi makineye bağlı anestezi &quot;maske&quot; içine fare burun yerleştirin. Maske bir 1 ml tek kullanımlık plastik pipet ucu (filtre) olmadan yapılmış ve dar uç (Şekil 2a, b) ile anestezi tüpe bağlanır. </li><li> Yavaşça parmağı ve başparmak serbest elin ve &quot;pop&quot; iyi pozlama ve kolay erişim (Şekil 2c) için dışarı göz kullanılarak nakledilmesi zorunluluğu göz göz kapakları çekin. Bu aşırı basınç tarafından fare nefes engel olmadan mükemmelleştirmek için biraz pratik gerektirirBoyun veya kafa kan akışını engelliyor. </li><li> Bir tek insülin şırınga (29 - 31G) kullanarak neşter gibi, dikkatli korneada sadece ucu nüfuz ve tek lateral insizyon yapın. ACE (Şekil 2d) dışına enjeksiyonu sırasında adacık reflü aza indirmek için kornea ve limbusun apeks arasındaki orta noktada kesi olun. </li><li> Dikkatle kesiden kanül (adacıkları ile önceden yüklenmiş) takın. </li><li> Yavaşça kanül adacıkları dışarı çıkarmak ve iris üstüne onları yatırmak. ACE aşırı basınç birikmesi nedeniyle adacık reflü önlemek için, kesi kadranda tersi mümkün olduğunca az ses (ler) olarak kısa bindirmeler içinde adacıklar çıkarın. Bu da kanül (adım 1.6 bakınız) yüklenirken tüp adacıkları sıkıştırma tarafından sağlanabilir. </li><li> Yavaşça ACE kanül dışarı çekin. Adacık büyük hacimli bir pres nedeniyle adacık reflüsü olarak enjekte edildi Bu, özellikle, önemli bir adımdırEmin ACE içinde biriken kaçınılmaz olabilir. Kanül etrafında kesiden fazla basıncı tahliye etmek ACE içinde iken adacık reflü aza indirmek / ortadan kaldırmak için, yavaşça kanül döndürün. Eğer kanül geri ve gerekirse, ACE tamamen geri çekilmeden önce basınç azalana kadar kanül beklemek için bir girişim olarak reflü belirtileri kontrol edin. </li><li> Steril PBS veya serum fizyolojik ile nakledilen gözü yıkayınız. </li><li> Ilk 48 saat boyunca post-operatif analjezi (subkutan 0.05-0.1 mg / kg) için buprenorfin enjekte edilir. </li><li> Hemen nakli sonrası nakledilen gözle eritromisin oftalmik antibiyotik merhem uygulayın. </li><li> Derlenme izin ısıtılmış bir kafesine geri hayvan yerleştirin. </li></ol>

<ol>
	<li>Weigert, R., Sramkova, M., Parente, L., Amornphimoltham, P., Masedunskas, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravital+microscopy:+a+novel+tool+to+study+cell+biology+in+living+animals.">Intravital microscopy: a novel tool to study cell biology in living animals.</a> Histochem. Cell Biol. 133 (5), 481-491 (2010).</li><li>Leibiger, I. B., Caicedo, A., Berggren, P. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+in+vivo+imaging+of+pancreatic+?-cell+function+and+survival+-+a+perspective.">Non-invasive in vivo imaging of pancreatic ?-cell function and survival - a perspective.</a> Acta Physiol. (Oxf). , (2011).</li><li>Wang, Y., Maslov, K., Kim, C., Hu, S., Wang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrated+photoacoustic+and+fluorescence+confocal+microscopy.">Integrated photoacoustic and fluorescence confocal microscopy.</a> IEEE Trans Biomed. Eng. 57 (10), 2576-2578 (2010).</li><li>Ntziachristos, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Going+deeper+than+microscopy:+the+optical+imaging+frontier+in+biology.">Going deeper than microscopy: the optical imaging frontier in biology.</a> Nat. Methods. 7, 603-614 (2010).</li><li>Aswathy, R. G., Yoshida, Y., Maekawa, T., Kumar, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+quantum+dots+for+deep+tissue+imaging.">Near-infrared quantum dots for deep tissue imaging.</a> Anal. Bioanal Chem. 397 (4), 1417-1435 (2010).</li><li>Ghoroghchian, P. P., Therien, M. J., Hammer, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+fluorescence+imaging:+a+personal+perspective.">In vivo fluorescence imaging: a personal perspective.</a> Wiley Interdiscip Rev. Nanomed Nanobiotechnol. 1 (2), 156-167 (2009).</li><li>Prescher, A., Mory, C., Martin, M., Fiedler, M., Uhlmann, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+FTY720+treatment+on+postischemic+pancreatic+microhemodynamics.">Effect of FTY720 treatment on postischemic pancreatic microhemodynamics.</a> Transplant Proc. 42 (10), 3984-3985 (2010).</li><li>Leblond, F., Davis, S., Vald&#233;s, P., Pogue, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-clinical+whole-body+fluorescence+imaging:+Review+of+instruments,+methods+and+applications.">Pre-clinical whole-body fluorescence imaging: Review of instruments, methods and applications.</a> J. Photochem. Photobiol. B. 98 (1), 77-94 (2010).</li><li>Toso, C., Vallee, J. P., Morel, P., Ris, F., Demuylder-Mischler, S., Lepetit-Coiffe, 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=Clinical+magnetic+resonance+imaging+of+pancreatic+islet+grafts+after+iron+nanoparticle+labeling.">Clinical magnetic resonance imaging of pancreatic islet grafts after iron nanoparticle labeling.</a> Am. J. Transplant. 8 (3), 701-706 (2008).</li><li>Denk, W., Strickler, J. H., Webb, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+laser+scanning+fluorescence+microscopy.">Two-photon laser scanning fluorescence microscopy.</a> Science. 248 (4951), 73-76 (1990).</li><li>Wang, B. G., Konig, K., Halbhuber, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+microscopy+of+deep+intravital+tissues+and+its+merits+in+clinical+research.">Two-photon microscopy of deep intravital tissues and its merits in clinical research.</a> J. Microsc. 238 (1), 1-20 (2010).</li><li>Denk, W., Delaney, K. R., Gelperin, A., Kleinfeld, D., Strowbridge, B. W., Tank, D. W., 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=Anatomical+and+functional+imaging+of+neurons+using+2-photon+laser+scanning+microscopy.">Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy.</a> J. Neurosci. Methods. 54 (2), 151-162 (1994).</li><li>Cahalan, M. D., Parker, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choreography+of+cell+motility+and+interaction+dynamics+imaged+by+two-photon+microscopy+in+lymphoid+organs.">Choreography of cell motility and interaction dynamics imaged by two-photon microscopy in lymphoid organs.</a> Annu. Rev. Immunol. 26, 585-626 (2008).</li><li>Khorshidi, M. A., Vanherberghen, B., Kowalewski, J. M., Garrod, K. R., Lindstrom, S., Andersson-Svahn, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+transient+migration+behavior+of+natural+killer+cells+imaged+in+situ+and+in+vitro.">Analysis of transient migration behavior of natural killer cells imaged in situ and in vitro.</a> Integr. Biol. (Camb). 3 (7), 770-778 (2011).</li><li>Matheu, M. P., Cahalan, M. D., Parker, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunoimaging:+studying+immune+system+dynamics+using+two-photon+microscopy.">Immunoimaging: studying immune system dynamics using two-photon microscopy.</a> Cold Spring Harb. Protoc. 2011, pdb.top99 (2011).</li><li>Celli, S., Albert, M. L., Bousso, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+the+innate+and+adaptive+immune+responses+underlying+allograft+rejection+by+two-photon+microscopy.">Visualizing the innate and adaptive immune responses underlying allograft rejection by two-photon microscopy.</a> Nat. Med. , (2011).</li><li>Fan, Z., Spencer, J., Lu, Y., Pitsillides, C., Singh, G., Kim, P., 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=In+vivo+tracking+of+'color-coded'+effector,+natural+and+induced+regulatory+T+cells+in+the+allograft+response.">In vivo tracking of 'color-coded' effector, natural and induced regulatory T cells in the allograft response.</a> Nat. Med. 16 (6), 718-722 (2010).</li><li>Sabek, O., Gaber, M. W., Wilson, C. M., Zawaski, J. A., Fraga, D. W., Gaber, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+of+human+islet+vascularization+using+a+dorsal+window+model.">Imaging of human islet vascularization using a dorsal window model.</a> Transplant Proc. 42 (6), 2112-2114 (2010).</li><li>Coppieters, K., Martinic, M. M., Kiosses, W. B., Amirian, N., von Herrath, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+technique+for+the+in+vivo+imaging+of+autoimmune+diabetes+development+in+the+pancreas+by+two-photon+microscopy.">A novel technique for the in vivo imaging of autoimmune diabetes development in the pancreas by two-photon microscopy.</a> PLoS One. 5 (12), e15732 (2010).</li><li>Martinic, M. M., von Herrath, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+imaging+of+the+pancreas+during+development+of+diabetes.">Real-time imaging of the pancreas during development of diabetes.</a> Immunol Rev. 221, 200-213 (2008).</li><li>Mostany, R., Portera-Cailliau, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Method+for+2-Photon+Imaging+of+Blood+Flow+in+the+Neocortex+through+a+Cranial+Window.">A Method for 2-Photon Imaging of Blood Flow in the Neocortex through a Cranial Window.</a> J. Vis. Exp. (12), e678 (2008).</li><li>Palmer, G. M., Fontanella, A. N., Shan, S., Hanna, G., Zhang, G., Fraser, C. L., 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=In+vivo+optical+molecular+imaging+and+analysis+in+mice+using+dorsal+window+chamber+models+applied+to+hypoxia,+vasculature+and+fluorescent.">In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent.</a> 6 (9), 1355-1366 (2011).</li><li>Jain, R. K., Munn, L. L., Fukumura, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissecting+tumour+pathophysiology+using+intravital+microscopy.">Dissecting tumour pathophysiology using intravital microscopy.</a> Nat. Rev. Cancer. 2 (4), 266-276 (2002).</li><li>Taylor, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+response+of+capillary+endothelium+to+changes+in+intravascular+pressure,+as+seen+in+the+rabbit's+ear+chamber.">The response of capillary endothelium to changes in intravascular pressure, as seen in the rabbit's ear chamber.</a> Aust. J. Exp. Biol. Med. Sci. 31 (5), 533-543 (1953).</li><li>Shan, S., Sorg, B., Dewhirst, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+rodent+mammary+window+of+orthotopic+breast+cancer+for+intravital+microscopy.">A novel rodent mammary window of orthotopic breast cancer for intravital microscopy.</a> Microvasc. Res. 65 (2), 109-117 (2003).</li><li>Speier, S., Nyqvist, D., Cabrera, O., Yu, J., Molano, R. D., Pileggi, A., 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=Noninvasive+in+vivo+imaging+of+pancreatic+islet+cell+biology.">Noninvasive in vivo imaging of pancreatic islet cell biology.</a> Nat. Med. 14 (5), 574-578 (2008).</li><li>Speier, S., Nyqvist, D., Kohler, M., Caicedo, A., Leibiger, I. B., Berggren, P. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+high-resolution+in+vivo+imaging+of+cell+biology+in+the+anterior+chamber+of+the+mouse+eye.">Noninvasive high-resolution in vivo imaging of cell biology in the anterior chamber of the mouse eye.</a> Nat. Protoc. 3 (8), 1278-1286 (2008).</li><li>Falck, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Site+of+production+of+oestrogen+in+the+ovary+of+the+rat.">Site of production of oestrogen in the ovary of the rat.</a> Nature. 184, 1082 (1959).</li><li>Bickford-Wimer, P., Granholm, A. C., Bygdeman, M., Hoffer, B., Olson, L., Seiger, A., 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=Human+fetal+cerebellar+and+cortical+tissue+transplanted+to+the+anterior+eye+chamber+of+athymic+rats:+electrophysiological+and+structural+studies.">Human fetal cerebellar and cortical tissue transplanted to the anterior eye chamber of athymic rats: electrophysiological and structural studies.</a> Proc. Natl. Acad. Sci. U.S.A. 84 (16), 5957-5961 (1987).</li><li>Adeghate, E., Donath, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+findings+in+long-term+pancreatic+tissue+transplants+in+the+anterior+eye+chamber+of+rats.">Morphological findings in long-term pancreatic tissue transplants in the anterior eye chamber of rats.</a> Pancreas. 5 (3), 298-305 (1990).</li><li>Abdulreda, M. H., Faleo, G., Molano, R. D., Lopez-Cabezas, M., Molina, J., Tan, 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=High-resolution,+noninvasive+longitudinal+live+imaging+of+immune+responses.">High-resolution, noninvasive longitudinal live imaging of immune responses.</a> Proc. Natl. Acad. Sci. U.S.A. , (2011).</li><li>Unutmaz, D., Xiang, W., Sunshine, M. J., Campbell, J., Butcher, E., Littman, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+primate+lentiviral+receptor+Bonzo/STRL33+is+coordinately+regulated+with+CCR5+and+its+expression+pattern+is+conserved+between+human+and+mouse.">The primate lentiviral receptor Bonzo/STRL33 is coordinately regulated with CCR5 and its expression pattern is conserved between human and mouse.</a> J. Immunol. 165 (6), 3284-3292 (2000).</li><li>Pileggi, A., Molano, R. D., Berney, T., Cattan, P., Vizzardelli, C., Oliver, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+induction+in+islet+cells+results+in+protection+from+apoptosis+and+improved+in+vivo+function+after+transplantation.">Heme oxygenase-1 induction in islet cells results in protection from apoptosis and improved in vivo function after transplantation.</a> Diabetes. 50 (9), 1983-1991 (2001).</li></ol>

PO.B. ticari bir hizmet platformu olarak gözün ön kamara kullanmak için gidiyor biyoteknoloji şirketi Biocrine, kurucularından biridir. AC Bu teknoloji koruma patent üzerinde.

Murin pankreas adacıklarının daha önce anlatıldığı gibi 33, yoğunluk gradient üzerinde saflaştırma, ardından kollajenaz sindirimi ile izole edilmiştir. İzole adacık nakli öncesi gecede kültüre edildi. Bu gerekli olmasa da, bu adacıkları izolasyon prosedürü kurtarmak için izin tavsiye edilir. O / sağlam adacık hayatta nakli sağlayacak gibi bu nakli diyabetik alıcılarında yapıldığında önemlidir. Transplantasyon oksijen / izofluran karışımı (% 1...

Intravital mikroskopi Gelişmeler vitro çalışmalar 1 öngördüğü olmayan fizyolojik fenomenler ortaya koymuştur. Bu canlı hayvan içine in vitro yöntemler konvansiyonel ile elde edilen bulgular çevirisinde meydan vurgulamaktadır. Son on yıl içinde, canlı hayvanlarda dokuların görselleştirme ölçüde görüntüleme yöntemleri 2, 3, 4, 5, 6, teknolojik gelişmeler tarafından geliştirildi. Bu non-invaziv, hedef dokuların boyuna görselleştirme sağlamak için deneysel hayvan modellerinde uygulanabilir uygulama ile in vivo görüntüleme yaklaşımlar ihtiyaç mahmuzlu vardır. Manyetik rezonans görüntüleme ve pozitron emisyon tomografisi veya biyolüminesans görüntüleme teknikleri vücut 7-8, 9 içinde derin organ / dokuların non-invaziv görüntüleme sağlamıştır. Ama bu tekniklerin kullanımı o rağmen, yüksek arka sinyaller ve düşük uzaysal çözünürlüğü nedeniyle hücre çözünürlüklü tek elde edemezf yüksek kontrast maddeler veya doku-spesifik lüminesans 4. Bu iki foton floresans konfokal mikroskopi 10 gelişiyle birlikte ele alındı. İki foton mikroskopi intravital görüntüleme çalışmaları görselleştirmek ve görülmemiş detayları 11, 12 ile hücresel olayları ölçmek için etkin. Bu sağlık ve hastalık 13, 14, 15, 16 kilit biyolojik süreçlerin karakterizasyonu yol açmıştır. Öncü intravital görüntüleme çalışmalarında öncelikle eksize edilen doku (örneğin lenf düğümleri) in vivo koşullarda &quot;taklit&quot; olmasına rağmen, diğer çalışmalar yerinde 17, 18, ​​19, 20, 21 görüntü maruz hedef doku invaziv yaklaşımlar kullanmışlardır. Diğer çalışmalar da invaziv yaklaşımlar ve in vivo 22, 23, 24, 25 sınırlı görüntüleme çözünürlüğü ile ilişkili sınırlamaları aşmak için &quot;pencere odacık modeller&quot; kullandık. Pencere oda modelinde, bir saydam bir pencere ile bir bölme cerrahi yönün azından deri içine implantehayvan kira konumları (dorsal veya kulak deri, meme yağ yastığı, karaciğer, vb) (örneğin fare, sıçan, tavşan). Bu yaklaşım açıkça in vivo görüntüleme yüksek çözünürlüklü sağlarken, bunu odasına implante bir invaziv cerrahi gerektirir ve birkaç hafta veya ay 22&#39;nin üzerinde uzunlamasına görüntüleme çalışmalarında karşılamak mümkün olmayabilir. Son zamanlarda gözün ön odasına yani minimal invaziv bir prosedür nakli ile yüksek çözünürlüklü konfokal mikroskopi birleştirerek (ACE) gibi bir &quot;doğal vücut penceresi&quot; sağladığı gösterilmiştir güçlü ve in vivo görüntüleme platformu 26, 27 çok yönlü. ACE içine Transplantasyon dokular 28, 29, 30, çeşitli biyolojik yönlerini incelemek için son birkaç yıldır kullanılmakta olan ve yüksek çözünürlüklü görüntüleme ile yeni kombinasyonu tek hücre-çözünürlük ile pankreas adacıklarının fizyolojisi okuyan etkin olmayan invazif ve uzunlamasına &lt;&gt; 26, 27 sup. Bu yaklaşım, hayvan modellerinde tip 1 diyabet (yayınlanmamış veri) geliştirilmesi sırasında otoimmün yanıtları incelemek için kullanılmıştır. Aynı zamanda, sırasıyla, ACE pankreas tomurcuklar ya da kişisel böbrek glomerülleri, (yayınlanmamış veriler) içine taşımak tarafından böbrek fonksiyonunu çalışmalarda, pankreas gelişme çalışma, hem de kullanılmıştır. Bu yaklaşımı kullanarak yeni bir rapor daha pankreas adacık nakli 31 sonrası bağışıklık yanıtlarını okumak uygulaması gösterilmiştir. Önemlisi, bu çalışma gerçekleştirmek için doğal bir vücut pencere sağlar gözün ön kamara içine nakli gösterdi: in vivo nakledilen dokuların (1) boyuna, non-invaziv görüntüleme, (2), in vivo hücresel fenotip ve uygulanabilirliğini değerlendirmek için cytolabeling in situ; hedef dokuda immün hücrelerinin infiltre (3) gerçek zamanlı izleme ve topikal uygulama veya göziçi enjeksiyon (4) yerel müdahale. Burada, dpankreas adacıkları kullanılarak gözün ön kamara içine nakli gerçekleştirmek için nasıl emonstrate.

<table border="1"><tbody><tr><td> Reaktif Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Açıklama / Yorumlar </td></tr><tr><td> IsoTHESIA (İzofluran) </td><td> Buttler Hayvan Sağlığı Kaynağı </td><td> 11695-6775-2 </td><td> % 99.9 İzofluran / ml </td></tr><tr><td> Ketaset (Ketamin HCL) </td><td> Fort Dodge Hayvan Sağlığı </td><td> 0856-2013-01 </td><td> Alternatif enjektabl anestezi </td></tr><tr><td> Beprenex (Buprenorfin HCL) </td><td> Reckitt Benckiser, Sağlık (UK) Ltd </td><td> 12496-075-7-1 </td><td> 0.3 mg / ml &#39; </td></tr><tr><td> Eritromisin oftalmik merhem, USP,% 0.5 </td><td> Akron </td><td> 17478-070-35 </td><td> Nakledilen göze profilaktik Uygulamalı </td></tr><tr><td> % 0.9 Sodyum Klorür (Tuz) </td><td> Hospira Inc </td><td> 0409-7983-03 </td><td> Iv enjeksiyon için. Steril </td></tr><tr><td> PBS </td><td> Gibco </td><td> 10010-023 </td><td> 1X. Steril </td></tr><tr><td> CMRL orta 1066 </td><td> Cellgro </td><td> 98-304-CV </td><td> Desteklenmiş, CIT modifikasyonu. Adacıklar için tercih edilen medya </td></tr></tbody></table>

transplantation into the anterior chamber of the eye for longitudinal, non-invasive in vivo imaging with single-cell resolution in real-time

Intravital görüntüleme biyolojik araştırma vazgeçilmez bir araç olarak ortaya çıkmıştır. Bu süreç içinde bir çok görüntüleme teknikleri invaziv olmayan hayvanlarda çeşitli biyolojik süreçlerin çalışma geliştirilmiştir. Ancak, mevcut intravital görüntüleme yöntemlerinin önemli bir teknik sınırlama tek hücreli çözünürlük yetenekleri ile non-invaziv, boyuna görüntüleme birleştirmek için yetersizliğidir. Biz gözün ön kamara içine nakli in vivo hücresel çözünürlüğü ile non-invaziv, boyuna görüntüleme sağlayan çok yönlü bir deney platformu sunan böyle önemli sınırlama circumvents nasıl burada gösterir. Biz fare nakli prosedürü göstermek ve yani klinik önemi, pankreas adacık nakli ile bir model kullanarak temsilcisi sonuçlar sağlar. Gözün anteryor odacık içine transplante dokuların çeşitli direkt görüş sağlamak için ek olarak, bu yaklaşım kayşat için bir platform sağlargerçekleştirerek n ilaçların uzun süreli takip ve hedef dokularda izleme. Çünkü gözü faydaları transplantasyon tedavileri sadece ön odasına, çok yönlülüğü, doku / hücre nakli, sinyal iletimi ve kanser veya otoimmün hastalık gelişme olarak fizyolojik ve patofizyolojik süreçlerin çalışma vivo uygulamalarda diğer uzanır.

&quot;Iyi&quot; bir transplantasyon tanımlayan birkaç parametre vardır. İyi bir transplantasyon video görülebileceği gibi kesi zaman kanama olmadan ilerler bir yoldur. Kanama ACE (Şekil 3a) içine neşter (iğne) sadece ucu nüfuz ederek önlenebilir / minimize edilmiştir. Bu da iris temas ve ponksiyon önlemeye yardımcı olacaktır. Ayrıca zaman (Şekil 3c, d) üzerinde kornea bulanıklıkları neden olmadan çok iyi iyileşir küçük bir kesi sağlayacaktır. Başarılı b...

Watch this Scientific Journal Video about Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time at JoVE.com

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Intraoküler nakli ve konfokal mikroskopi birleştirerek yeni bir yaklaşım aşılı dokuların içinde tek hücreli çözünürlüğe sahip uzunlamasına, non-invaziv, gerçek zamanlı görüntüleme sağlar<em&gt; In vivo</em&gt;. Biz fare gözün ön kamaraya pankreas adacık nakli nasıl gösterilmektedir.

Boyuna, Non-invaziv için Eye Ön Kamara içine Nakli<em&gt; In vivo</emGerçek zamanlı Tek hücreli Çözünürlük&gt; Görüntüleme

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

transplantation-into-anterior-chamber-eye-for-longitudinal-non

Research

JoVE Journal

Medicine

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

13.3K Görüntüleme.  University of Miami Miller School of Medicine. Intraoküler nakli ve konfokal mikroskopi birleştirerek yeni bir yaklaşım aşılı dokuların içinde tek hücreli çözünürlüğe sahip uzunlamasına, non-invaziv, gerçek zamanlı görüntüleme sağlar In vivo. Biz fare gözün ön kamaraya pankreas adacık nakli nasıl gösterilmektedir.

Video: Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in functional parameters of key clinical significance. Two-dimensional movies were acquired at high frame rate (8 kHz) and were utilized to estimate high-quality myocardial strain. Two-dimensional elastograms (strain images), as well as strain profiles, were visualized. Results were powerful in quantitatively assessing IR-induced changes in cardiac events including left-ventricular (LV) contraction, LV relaxation and isovolumetric phases of both pre-IR and post-IR beating hearts in intact mice. In addition, compromised sector-wise wall motion and anatomical deformation in the infarcted myocardium were visualized.  The elastograms were uniquely able to provide information on the following parameters in addition to standard physiological indices that are known to be affected by myocardial infarction in the mouse: internal diameters of mitral valve orifice and aorta, effective regurgitant orifice, myocardial strain (circumferential as well as radial), turbulence in blood flow pattern as revealed by the color Doppler movies and velocity profiles, asynchrony in LV sector, and changes in the length and direction of vectors demonstrating slower and asymmetrical wall movement. This work emphasizes on the visual demonstration of how such analyses are performed. 

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart 

High frequency Doppler ultrasound is a novel technology for assessing regional myocardial function. This work presents first evidence demonstrating applicability of this versatile imaging platform for the repeated measure of myocardial strain, dp/dt, and mitral regurgitation in the ischemia-reperfused (IR) murine heart.

Yüksek frekanslı Yüksek çözünürlüklü Ekokardiyografi: İskemi-Reperfüze murin Kalp Miyokard Strain, kontraktilite ve Mitral Yetmezliğinde Non-invaziv Tekrarlanan Ölçüm İlk Kanıt

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in ...

Tıp

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

Jinekolojik Onkoloji Sentinel lenf nodu intraoperatif Algılama için Multispektral Gerçek zamanlı Floresans Görüntüleme

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

Sıçan Böbrek Nakli sonra Akut Allogreft Reddi non-invaziv Görüntüleme Kullanımı<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

Ses tabanlı Sanal Oyun Ortamı Geliştirilmesi Kör Navigasyon Becerileri ile Yrd etmek

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an increased risk of vascular morbidity.&#160;Yet, albeit&#160;the mounting clinical evidence, there is a paucity of clear studies of vascular toxicity and therefore the etiology of a heterogeneous group of vascular/cardiovascular disorders remains to be elucidated. Moreover, the mechanism that may underlie vascular toxicity can completely differ from the principles of chemotherapy-induced cardiotoxicity, which is related to direct myocyte injury. We have established a real-time, in vivo molecular imaging platform&#160;to evaluate&#160;the potential&#160;acute&#160;vascular toxicity of anti-cancer therapies.
We have set up a platform of in vivo, high-resolution molecular imaging&#160;in mice,&#160;suitable for visualizing vasculature within confined organs and reference blood vessels within the same individuals whereas each individual serve as its own control. Blood vessel walls were impaired after doxorubicin administration, representing a unique mechanism of vascular toxicity that may be the early event in end-organ injury. Herein, the method of fibered confocal fluorescent microscopy (FCFM) based imaging is described, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Kemoterapi kaynaklı Vasküler Toksisite - Gerçek zamanlı<em&gt; In vivo</em&gt; Gemi Düşüklüğü Görüntüleme

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Canlı Fareler Seri endoskopik Görüntüleme kullanma Bağırsak Patolojilerde Yeni Therapeutics Etkinliğinin Non-invaziv Değerlendirme

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

Yağ Dokusu Depolarında içine Nakli Enkapsülasyon termojenik Preadipositler

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.

This protocol describes the use of multiphoton microscopy to perform extended time-lapse imaging of multicellular interactions in real time, in vivo at single cell resolution.

Tümör mikroçevresinde Gerçek zamanlı Multisellüler Dynamics Genişletilmiş Time-lapse intravital Görüntüleme

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and ...

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular plasticity or damage occurring overtime and in relation to neuronal activity or blood flow. In vivo two-photon microscopy allows the study of the structural and functional plasticity of large cellular units in the living brain. In particular, the thinned-skull window preparation allows the visualization of cortical regions of interest (ROI) without inducing significant brain inflammation. Repetitive imaging sessions of cortical ROI are feasible, providing the characterization of disease hallmarks over time during the progression of numerous CNS diseases. This technique accessing the pial structures within 250 &#956;m of the brain relies on the detection of fluorescent probes encoded by genetic cellular markers and/or vital dyes. The latter (e.g., fluorescent dextrans) are used to map the luminal compartment of cerebrovascular structures. Germane to the protocol described herein is the use of an in vivo marker of amyloid deposits, Methoxy-O4, to assess Alzheimer&#39;s disease (AD) progression. We also describe the post-acquisition image processing used to track vascular changes and amyloid depositions. While focusing presently on a model of AD, the described protocol is relevant to other CNS disorders where pathological cerebrovascular changes occur.

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

This manuscript describes a procedure to track the remodeling of the cerebrovasculature during amyloid plaque accumulation in vivo using longitudinal two-photon microscopy. A thinned-skull preparation enables the visualization of fluorescent dyes to assess the progression of cerebrovascular damage in a mouse model of Alzheimer&#39;s disease.

uzunlamasına<em&gt; İn Vivo</em&gt; Cerebrovasculature Görüntüleme: MSS Hastalıkları uygunluk

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

Yeniden Arterialized sıçan kısmi karaciğer nakli bir vivo içinde gemi odaklı % 70 ile Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...