우리는 Drs을 인정합니다. Camillo Ricordi, Antonello Pileggi, R. Damaris Molano, 스테판 Speier 유익한 토론 다니엘 Nyqvist. 우리는 또한 비디오 녹화와 관련하여 도움이 Eleut 헤르 난 데스와 디에고 에스피노사 - Heidmann 기술 지원 및 마이크 발데스와 마가렛 Formoso 감사드립니다. 바이런 말도 나도는 편집 기록, 최종 비디오를 제작했다. 연구 지원은 당뇨병 연구소 재단 (에서 제공 한 <a href="http://www.DiabetesResearch.org" target="_blank">www.DiabetesResearch.org</a> ), NIH / NIDDK / NIAID (MHA에 F32DK083226, AC에 대한 NIH RO3DK075487, PO.B.에 U01DK089538). PO.B에 대한 추가 연구 지원은 Karolinska Institutet, 스웨덴 연구위원회, 스웨덴 당뇨병 재단, 가족 Erling - 페르손 재단, 가족 Knut와 앨리스 발렌 버그 재단, 활기찬 Skandia 보험 회사 (주)에서 자금을 통해 제공되었다 ( Karolinska Inst에서 당뇨병의 FP7 - 228933-2), 전략적 연구 프로그램itutet, 노보 Nordisk 재단, 그리고 부두 폰 Kantzow의 재단.

다음 절차는 2 단계로 입체경에서 수행되며, 첫 번째 단계는 캐뉼라로 섬을로드 포함하고 두 번째 단계는 ACE에 실제 이식입니다. 동물에서 수행 모든 절차는 마이애미 대학의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 승인되었습니다. 1. 이식을위한 정맥의로드 섬 <ol><li> 축소 서클 요리를 회전하여 센터 배양 접시에있는 섬입니다. </li><li> &quot;저수지&quot;에서 캐뉼라를 분리하고 깨끗한 표면에 캐뉼라와 연결 튜브를 놓습니다. 저장소는 필터 (그림 1a)없이 300 μl 일회용 플라스틱 피펫 팁으로 만든 할 수 있습니다. </li><li> 저수지에 aspirating 때 섬의 지속적인 흐름을 보장하기 위해 저수지에서 기포를 제거. 저수지를 플러싱하는 것은 발 페달을 사용하여 핸즈프리 동력 주사기 - 드라이버를 앞으로 운전하면됩니다(그림 1b, C). 이것은 또한 저수지 (예 : 생리, PBS 또는 문화 미디어로 살균 솔루션으로 사전로드)로 섬의 열망을 허용하도록 주사기에 공간을 할 것입니다. </li><li> 부드럽게 저수지에 섬의 원하는 금액을 대기음. 사람들이 저수지를 입력하고 하단을 향해 함께 남아있게됩니다 섬은 소용돌이 경향이 있습니다. 흡인이 발 페달을 사용하여 이전 버전과 동력 주사기 - 드라이버를 구동하여 이루어집니다. </li><li> 연결 튜브를 통해 저수지에 캐뉼라를 다시 연결하십시오. </li><li> 배양 접시에 다시 캐뉼라 팁을 놓고 캐뉼라로 다음 튜브에 저수지의 섬을 플러시. 당신은 부드럽게 (도청) 튜브 (그림 1D) &quot;을 flicking&quot;에 의해 튜브 / 캐뉼라를 백업 채울 때 섬이 함께 남아 있는지 확인합니다. 캐뉼라를 밖으로 내보냈 아르 앞서 섬의 모든 기포 전이나 후에 중지합니다. 확실하지 않은 경우, 섬이 남아 공기로 캐뉼라의 뒷면을 입력으로 중지앞서 섬의 거품은 ACE의 섬의 역류 (역류)을 방지 할 수 있습니다. 밤새 분산됩니다. </li><li> 이 단계에서, 당신은 ACE (다음 단계 참조)에 섬을 삽입 할 수 있습니다. </li></ol> 2. 눈의 앞쪽 상공 회의소에 작은 섬 이식 <ol><li> 입체경 아래 따뜻한 패드에 anesthetized 마우스를 배치합니다. </li><li> 산소 / isoflurane 마취 기계에 연결 마취 &quot;마스크&quot;에 마우스의 코를 놓습니다. 마스크 1 ML 일회용 플라스틱 피펫 팁 (필터없이)으로 만든과 좁은 끝 (그림 2A, B)를 통해 마취 튜브에 연결되어 있습니다. </li><li> 부드럽게 손가락과 엄지 손가락 무료 손과 &quot;팝업&quot;더 나은 노출과 접근이 용이합니다 (그림 2C)를 밖으로 눈을 사용하여 이식 할 눈의 눈 뚜껑을 철회. 이에 과도한 압력에 의해 마우스의 호흡을 방해하지 않고 완벽하게 연습을 필요로합니다목이나 머리에 혈액 흐름을 차단. </li><li> 일회용 인슐린 주사기 (29 - 31G)를 사용 메스로 신중하게 각막 만 끝을 관통하고 하나의 측면 절개를합니다. ACE (그림 2D)의 아웃 분사 동안 섬의 환류을 최소화하기 위해 각막과 limbus의 꼭대기 사이의 중간 지점에 절개를합니다. </li><li> 조심스럽게 절개를 통해 캐뉼라 (섬과 미리로드) 삽입합니다. </li><li> 천천히 캐뉼라의 섬을 추출하고 홍채의 상단에이를 입금. ACE에 과도한 압력 축적으로 인해 작은 섬 역류를 방지하려면 절개로 사분면의 맞은 편에 가능한 한 작은 볼륨 (들)로 한때 thrusts에있는 섬을 꺼냅니다. 이것은 더 캐뉼라 (단계 1.6 참조)로드하는 동안 튜브에 섬을 압축하여 보장 할 수 있습니다. </li><li> 천천히 ACE의 캐뉼라를 철회. 섬의 큰 볼륨 시장을 대표로 인해 작은 섬 환류로 주입 된 경우, 특히 중요한 단계입니다확인 ACE 내부에 구축 피할 수 있습니다. 캐뉼라 주위의 절개를 통해 과도한 압력을 출시 할 ACE 내부하면서 작은 섬 역류를 최소화 / 제거하려면 부드럽게 캐뉼라를 회전 할 수 있습니다. 당신이 캐뉼라를 철회하고, 필요한 경우, ACE의 완전히 retracting하기 전에 압력이 가라까지 캐뉼라를 기다려야 시도로 환류의 흔적이 있는지 확인합니다. </li><li> 멸균 PBS 또는 식염수에 이식 눈을 씻어. </li><li> 처음 48 시간에 대한 수술 진통제 (피하 0.05-0.1 밀리그램 / kg)에 buprenorphine를 삽입. </li><li> 바로 이식 후 이식 눈에 에리스로 마이신의 안과 항생제 연고를 적용 할 수 있습니다. </li><li> 마취에서 회복을 허용 할 수있는 가온 케이지에 다시 동물을 놓으십시오. </li></ol>

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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. 상용 서비스 플랫폼으로 눈의 앞쪽 챔버를 사용하는 것이다 생명 공학 회사 Biocrine의 창립자 중 하나입니다. AC는이 기술을 보호 특허에 있습니다.

손쥐 췌장 섬은 이전에 33 설명한 바와 같이, 밀도 기울기에 정화에 이어 collagenase의 소화를 사용하여 분리 하였다. 절연 섬은 이식 전에 하룻밤 배양 하였다. 이것이 필요하지 않을 수도 있지만,이 섬은 격리 절차에서 복구 할 수 있도록하는 것이 좋습니다. 가 / 강력한 섬을 생존의 이식을 보장하므로이 이식이 당뇨병 수신자에 수행 할 때 중요합니다. 이식은 산소 / i...

intravital 현미경의 발전은 체외 연구 1에 의해 예측하지 생리 현상을 보여 왔습니다. 이것은 살아있는 동물로 체외 방법에 기존하여 얻은 결과를 번역에 도전을 강조 표시합니다. 지난 10 년 간, 생활 동물 조직의 시각화는 상당히 이미징 modalities 2, 3, 4, 5, 6에 기술 진보에 의해 개선되었다. 이 비 invasively 대상 조직의 길이 방향 시각화를 사용하는 실험 동물 모델에서 가능한 응용 프로그램과 생체 이미징 접근에 대한 필요성을 가했다 있습니다. 이러한 자기 공명 영상 및 양전자 방출 단층 촬영이나 bioluminescence와 같은 이미징 기술은 신체 7-8, 9에서 깊은 장기 / 조직의 비 침습적 영상을 사용했습니다. 그러나 이러한 기술을 사용 O에도 불구하고, 높은 배경 신호와 낮은 공간 해상도로 인해 휴대 해상도 하나를 달성 할 수 없습니다F 고 대비 자료 나 조직 별 발광을 4. 이것은 두 광자 형광 공 촛점 현미경 (10)의 도래와 함께 해결되었습니다. 두 광자 현미경은 intravital 이미징 연구 시각화하고 전례가없는 세부 사항 11, 12로 휴대 이벤트를 정할 수있었습니다. 이 건강과 질병 13, 14, 15, 16의 주요 생물학적 과정의 특성을 이끌었습니다. 선구 intravital 이미징 연구가 주로 excised 조직 (예 : 림프절)의 생체 조건에서 &quot;없잖아?&quot;동안, 다른 연구는 시츄 17, 18, ​​19, 20, 21 이미지 노출 대상 조직에 침투 방법을 사용했습니다. 다른 연구는 또한 침습적 방법과 생체 22, 23, 24, 25 제한된 영상 해상도와 관련된 제한을 회피하기 위해 &#39;창 챔버 모델 &quot;을 사용했습니다. 창 챔버 모델에서 투명 창 챔버는 수술 diffe의 피부에 이식되어동물의 임대 위치 (등쪽이나 귀 피부, 유방 지방 패드, 간, 등) (예 : 마우스, 쥐, 토끼). 이 방법이 명확하게 생체 이미징에 높은 해상도를 수 있지만, 챔버 이식 해 침습적 수술을 필요하며 몇 주 또는 몇 달 22여 종 방향 이미징 연구를 수용 할 수없는 경우가 있습니다. 그것은 최근 눈의 앞쪽 챔버에, 즉 최소한 침략 절차, 이식과 고해상도 공 촛점 현미경을 결합하는 (ACE)로 &quot;자연 몸 창&quot;을 제공하는 입증 된 강력하고 생체 이미징 플랫폼 26, 27의 다양한. ACE에 이식이 조직 28, 29, 30의 다양한 생물학적 측면을 연구하기 위해 지난 몇 수십 년 동안 사용되었습니다, 그리고 고해상도 영상과의 최근 조합은 단일 셀 해상도로 췌장 섬의 생리를 공부 활성화 비 invasively 및 길이 방향 &lt;&gt; 26, 27 논의하게 될 것입니다. 이 방법은 동물 모델에서 제 1 형 당뇨병 (게시되지 않은 데이터)의 개발 기간 동안 면역 반응을 연구하는 데 사용되었습니다. 또한 각각 ACE의 췌장 꽃 봉오리 또는 개별 신장 glomeruli, (게시되지 않은 데이터)에 이식하여 신장 기능의 연구에서, 췌장 개발을 공부뿐만 아니라하는 데 사용되었습니다. 이 방법을 사용하는 최근의 보고서는 더 췌장 췌장 이식 31 일 이후 면역 반응을 연구하기위한 응용 프로그램을 보여 주었다. 중요한 것은,이 연구는 수행 할 천연 바디 창 제공하는 눈의 앞쪽 챔버에 그 이식을 보여 주었다 : 생체에 이식 된 조직 (1) 길이, 비 침습적 영상을, (2) 생체에서의 세포 표현형 및 생존 능력을 평가하기 위해 cytolabeling 현장, 대상 조직에 면역 세포에 침투 (3) 실시간으로 추적하고, 국소 응용 프로그램 또는 안구 내 주사로 (4) 지역 개입했습니다. 여기, 우리는 D췌장 섬을 사용하여 눈의 앞쪽 챔버 내로 이식을 수행하는 방법을 emonstrate.

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 설명 / 의견 </td></tr><tr><td> IsoTHESIA (Isoflurane) </td><td> Buttler 동물 건강 공급 </td><td> 11695-6775-2 </td><td> 99.9 % Isoflurane / ML </td></tr><tr><td> Ketaset (케타민 HCL) </td><td> 요새 닷지 동물 건강 </td><td> 0856-2013-01 </td><td> 대체 injectable 마취 </td></tr><tr><td> Beprenex (Buprenorphine HCL) </td><td> Reckitt Benckiser 건강 (UK) (주) </td><td> 12496-075-7-1 </td><td> 0.3 MG / ML </td></tr><tr><td> 에리스로 마이신의 안과 연고의 USP, 0.5 % </td><td> 애 크런 </td><td> 17478-070-35 </td><td> 이식 눈에 prophylactically 적용 </td></tr><tr><td> 0.9 %의 나트륨 염화물 (식염) </td><td> Hospira 주식회사 </td><td> 0409-7983-03 </td><td> IV 주입하십시오. 불모의 </td></tr><tr><td> PBS </td><td> Gibco </td><td> 10010-023 </td><td> 1X. 불모의 </td></tr><tr><td> CMRL 중간 1066 </td><td> Cellgro </td><td> 98-304 - CV </td><td> 보충, CIT의 수정. 섬에 대한 선호하는 미디어 </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 영상은 생물학적 연구에 필수적인 도구로 떠오르고있다. 이 과정에서 많은 이미징 기술이 아닌 invasively 동물에서 다른 생물학적 과정을 연구하기 위해 개발되었습니다. 그러나, 기존의 intravital 이미징 modalities의 주요 기술적 인 제한은 단일 셀 해상도 기능으로 비 침습적, 길이 방향의 영상을 결합 할 수 없다는 것입니다. 우리는 눈 앞 챔버 내로 이식은 생체 내 세포 해상도 비 침습적, 세로 이미지 수있는 다양한 실험 플랫폼을 제공하는 등 상당한 제한을 circumvents 방법을 여기에 표시됩니다. 우리는 마우스에 이식 절차를 설명하고, 즉 임상 관련성, 췌장 췌장 이식과 모델을 사용 대표적인 결과를 제공합니다. 눈의 앞쪽 챔버 내로 이식 조직의 다양한 직접 시각화를 지원하는 것 외에도,이 방법은 애추 할 수있는 플랫폼을 제공합니다수행하여 N 약물의 장기 후속 및 대상 조직의 모니터링. 때문에 눈 혜택 이식 요법뿐만 아니라 앞쪽에 챔버로의 다양성, 조직 / 세포 이식의, 이건 정말 신호 전달과 암이나 면역 질환 개발과 같은 생리와 pathophysiological 과정을 공부하고 생체 응용 프로그램에서 다른 확장합니다.

&quot;좋은&quot;이식을 정의하는 몇 가지 매개 변수가 있습니다. 좋은 이식은 비디오에서 볼 수 있듯이 절개 할 때 출혈이없이 진행이 하나입니다. 출혈은 ACE (그림 3A)에 메스 (바늘) 만 팁을 관통하여 예방 / 최소화합니다. 이 또한 홍채의 접촉과 주사를 방지하는 데 도움이됩니다. 또한 시간 (그림 3C, D)을 통해 각막의 변화를 유발하지 않고 잘 치유 될 소형 절개를 보장합니...

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

안구 이식 및 공 촛점 현미경을 결합 새로운 접근 방식은 이식 조직 내에서 단일 셀 해상도 세로, 비 침습적 실시간 이미징을 가능하게<em&gt; 생체 내</em&gt;. 우리는 마우스 눈의 앞쪽 챔버에 췌장 섬을 이식하는 방법을 보여줍니다.

길이, 비 침습적을위한 안구의 앞쪽 상공 회의소에 이식<em&gt; 생체에</em실시간으로 단일 셀 해상도&gt; 영상

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

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

13.3K 조회수.  University of Miami Miller School of Medicine. 안구 이식 및 공 촛점 현미경을 결합 새로운 접근 방식은 이식 조직 내에서 단일 셀 해상도 세로, 비 침습적 실시간 이미징을 가능하게 생체 내. 우리는 마우스 눈의 앞쪽 챔버에 췌장 섬을 이식하는 방법을 보여줍니다.

비디오: 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.

고주파 고해상도 Echocardiography : 국소 빈혈 - reperfused Murine 중심부에 심근 스트레인, 수축성 및 승모판 병리 현상의 비침습 반복 측정에 대한 최초의 증거

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

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의학

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.

Gynecologic 종양학의 센티널 림프절의 Intraoperative 감지 Multispectral 실시간 형광 이미징

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.

쥐 신장 이식 후 급성 동종 이식 거부의 비 침습 이미징 사용<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.

오디오 기반의 가상 게임 환경의 개발은 맹인의 탐색 기술을 지원하는

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.

화학 요법에 의한 혈관 독성 - 실시간<em&gt; 생체 내</em&gt; 선박 손상의 영상

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.

라이브 마우스의 직렬 내시경 이미징을 사용하여 소장 병변에 대한 새로운 치료제의 효능의 비 침습적 평가

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.

지방 조직의 종점에 이식 캡슐화 열 발생 Preadipocytes

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.

종양 미세 환경에서 실시간 다세포 역학의 확장 시간 경과 생체 내에 영상

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.

종<em&gt; 생체</em&gt;은 Cerebrovasculature의 이미징 : CNS 질환에 관련성

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.

다시는 vivo에서 선박 지향 70% 쥐 부분 간 이식 Arterialized Hepatectomy

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