우리는 전문적인 기술 지원을 소냐 Dufentester 및 엘크 웨버 감사합니다. 우리는 의료 정보학 지원을위한 원고와 스테판 브루크너를 교정에 대한 Faekah Gohar 감사합니다. 이 작품은 그렇지-크로네 -는 Fresenius 재단 (2012_A94)에서 학제 보조금에 의해 지원되었다. D. Bettenworth 의학 학부, Westfälische Wilhelms-Universität 뮌스터에서 연구 친교에 의해 지원되었다. M. 브루크너는 도이치 Forschungsgemeinschaft (DFG SFB1009B8)의 &quot;Gerok&quot;회전 위치에 의해 지원되었다. 우리는 마우스 만화의 그림은 헤이 케 블룸 감사합니다.

모든 동물 실험은 독일어 동물 보호법에 따라 Landesamt 엘리제 투르, UMWELT 싶게 Verbraucherschutz (LANUV)에 의해 승인되었다. 1 재료 및 실험 설정 <ol><li> 동물 보호 <ol><li> 20~25g 무게 균주의 여성 또는 남성 마우스를 사용하여 지역 동물 보호 법규에 따라이를 수용. </li><li> 설치류에 대한 특별 차우와 마우스를 공급하고 알팔파없는 우를 적용​​ 적어도 3 일 endoluminal 자동 형광을 최소화하기 위해 시험을에서 형광 전에. </li><li> 멸균 마시는 임의 량의 물을 제공합니다. </li></ol></li><li> 급성 DSS 유발 성 대장염의 유도 <ol><li> 물을 멸균 100 mL에 DSS의 3g을 용해시켜 용액 : 덱스 트란 황산 나트륨 (36,000-50,000 다 DSS, 분자량) (/ V w) 3 %를 준비합니다. 마우스 수의에 전용 식수로이 솔루션을 제공하고 마우스 / 일 당 DSS-용액 5 ㎖를 계산합니다. 공급 계속DSS 수의 9없이 멸균 물 ROL 마우스. </li></ol></li><li> 대장 암의 유도 <ol><li> 돌연변이 azoxymethane (AOM) 용​​해 (암과 유전 적 손상을 일으킬 수 있음주의!) 멸균 식염수에 1 mg / ml의 최종 농도를 얻었다. 체중이 복강 1 ML의 주사기 (30 G) (10)를 사용하여 kg 당 10 mg을 AOM의 단일 용량을 적용합니다. </li><li> 21 3퍼센트 7 일 0에서 (w / v)의 DSS, 14 일의 반복주기, 일 28-35 하루 42-49로 (제어 마우스 제외) 도전 마우스는 염증 구동 대장 cancerogenesis을 유도한다. 이들 과제 (자세한 시간 일정에 대한 그림 4A 참조) 사이에 멸균 물 쥐를 넣습니다. 실험을하는 동안 멸균 물을 제어 쥐를 넣습니다. </li></ol></li><li> 형광 내시경 검사의 준비 (FE) <ol><li> 사용 플루 오레 Isothiocyanat (FITC) - 덱스 트란 (분자량 70,000 다; FITC : 포도당 = 1 : 250)에 대한 dete이형성증 관련된 혈관 패턴의 시각 개선에 의한 대장 선종의 ction의. </li><li> 정맥 5 분 형광 내시경 검사 전에 100 ㎕의 PBS에 희석 60 밀리그램 FITC - 복합 덱스 트란을 관리 할 수​​ 있습니다. </li></ol></li><li> 마취 <ol><li> (; 1.5-2 부피 %의 이소 플루 란 [2 - 클로로 2 - (다이 플루오 로메 톡시) -1,1,1 - 트라이 플루오로 - 에탄] 1.5 2 LO / 분보기) 이소 플루 란 마취를 위해 연속적인 공급을 제공한다. 단단히 마취를 제어하기 위해 안면 마스크와 특수 동물 용 마취 장비를 사용합니다. </li></ol></li><li> 관장의 준비 <ol><li> 유체 관장 2 ㎖ 주입 (내용 : 나트륨이 수소에게 (/ w 1.5 %를 V) 및 나트륨 / v)의 와트 (11 % 인산이 수소) 결장에 중요한 배설물 적재가 의심되는 경우보기를 모호하게 할 수있다. </li></ol></li></ol> 2 기술 장비 <ol><li> 개발 및 작은 동물 내시경의 사용 승인 된 동물 내시경 워크 스테이션을 사용합니다. 우와 연결카메라 유닛, 크세논 광원, 공기 펌프 및 백색광 내시경 용 종래의 PC 모니터에 rkstation. 그런 다음 카메라와 소형 단단한 망원경 (; 그림 5 1.9 mm 외경 10cm 길이)를 연결합니다. </li><li> 생검 겸자 또는 주입 관의 응용 프로그램에 대해 작동 채널 (그림 5D)와 내시경 칼을 사용합니다. 진단 대장 내시경 검사를위한 채널을 작동하지 않고 외피를 사용합니다. </li><li> 사용되는 트레이서를 여기시키기 위해 형광 내시경 광원의 설정을 구성 (예를 들면, FITC-덱스 트란에 대한 공역 내지 490). 또한, 망원경 카메라 (FITC-결합 된 덱스 트란에 대한 예를 들면, 525 nm의) 사이의 적절한 대역 통과 필터를 통합 할 수 있습니다. </li><li> 조직 검사를받을 수있는 내시경의 작업 채널을 통해 유연한 생검 겸자 (3 차르., 28cm)를 삽입합니다. </li><li> 국소, 점막 내암의 작업 채널을 통해 유연한 주입 관 (0.96 mm)를 소개하거나를 endoluminal진단 또는 치료제의 dministration. </li><li> 42 ° C의 온도로 가열 가능한 시험 테이블을 사용합니다. 이 마우스는 시험 기간 동안 저체온되는 것을 방지합니다. </li></ol> 동물의 3 마취 <ol><li> 작지만 누액 방지 상자에 마우스를 놓고 이소 플루 란을 관리 (100 % (v / v)로, 5 부피 % / 분 3 L). 마우스가 의식을 잃을 때까지 기다립니다. </li><li> 내시경 검사를위한 시험 테이블에 마우스를 전송합니다. 용량 100 %의 v / V, 1.5 부피 %, / 분 1.5 L와 얼굴에 마스크를 통해 이소 플루 란 흡입을 계속합니다. 항상 마취 동안 눈 건조를 방지하기 위해 눈 연고를 적용합니다. </li><li> 반사 신경을 확인하여 마취의 효과를 평가합니다. &#39;반사 주위에 회전을&#39;확인 : 충분히 마취 경우, 뒷면에 누워 마우스 돌아서는 안된다. 확인 &#39;발가락을 반사&#39;마취 부드러운 다리의 철회 (의 단계로 이어질 안 동물의 발가락 사이에서 곤란, 적절한 때수술 허용). </li></ol> 4 대장 내시경 검사 <ol><li> 시험 테이블에 해당 다시 / 마취 마우스 경향이 놓습니다. </li><li> 중요한 배설물로드가보기를 가릴 수 있다는 의심되는 경우 콜론으로 놨는데 캐 뉼러를 통해 관장 2 ㎖를 관리. 마우스가 관장을 투여 한 후 배변 때까지 기다립니다. 천공을 피하기 위해 매우 조심스럽게 내시경을 삽입합니다. </li><li> 그 중 하나는 에어 펌프에 연결되어있는 시스의 밸브를 모두 연다. 공기를 분배하기 위해 검지 손가락으로 다른 밸브를 밀봉합니다. 특히 조직 검사 또는 주사 가지 경우에, 천천히, 조심스럽게 공기와 콜론을 부풀려. </li><li> 구멍 (항문에서 4~5cm)를 피하기 위해 오른쪽 대장의 굴곡으로 만까지 내시경을 진행합니다. </li><li> 진단 대장 내시경 검사 <ol><li> 내시경을 다시 잡아 당기면서 염증이나 악성 변화의 점막을 검사합니다. 창자의 전체 둘레를 평가하기 위해 다시 천천히 잡아 당깁니다. 관내 연평균 평가필요에 따라 적절한 설립 채점 시스템을 사용 thologies. </li><li> 상처 영역의 반복적 인 시각화하는 동안 이미지 수집에 대해 동일 내시경의 위치를​​ 보장하기 위해, 쥐의 항문 점막 병변 사이의 거리를 확인합니다. 또한, 내시경 화상 획득 동안에 상처 영역 사이에 동일한 거리를 달성하기 위해 스페이서로서 생검 겸자의 끝을 사용한다. 상처의 크기가 3mm를 포함 내시경 시쓰의 크기와 관련된다. 참고 : 이전 시험의 사진 문서와 광학 비교하여 동일한 위치에 배치 내시경. 각에서 같은 각도와 거리에서 측정 병변은 내시경 검사를 따릅니다. </li></ol></li><li> 생검 절차 <ol><li> 이 연구자의 도움으로 조직 검사를 가져 가라. 핀셋의 선단이 제 조사자에 모니터에 표시 될 때까지 워킹 채널을 통해주의 깊게 생검 겸자를 소개한다. 신중하게 개폐 집게무효 천공. </li><li> 병리의 사이트에 집게를 이동합니다. </li></ol></li><li> 사출 절차 <ol><li> 이 연구자의 도움으로 주입 절차를 수행합니다. 완전히 제와 사전 채우기 유연한 주입 관 (0.96 mm)을 투여한다. 캐 뉼러 (30 G)이 두 번째 조사에 모니터에 표시 될 때까지 작업 채널을 통해 튜브를 밀어 넣습니다. 미세 주사기를 준비하고 조심스럽게 진단 또는 치료제의 요구 된 양을 관리 할 수​​ 있습니다. 사출 볼륨은 50 ㎕의 최대해야한다. </li><li> 15-30 도의 각도로 submocosa 바늘을 삽입한다. 점막의 방향으로 경사에 직면하고있다. 점막이 성공적으로 주입 후 특성 리프팅 기호를 보여줍니다. </li></ol></li><li> 형광 내시경 (FE) <ol><li> 정맥 전에 내시경 검사를 형광성 100 ㎕의 PBS에 희석 60 밀리그램 FITC - 복합 덱스 트란을 관리 할 수​​ 있습니다. </li><li> 주입 사이의 최적의 시점을 확인당신의 형광 표지 된 추적 및 추적 약리학에 의존하는 이미지를 만드는 절차. 사용되는 추적의 여기 및 발광 파장에 따라 대역 통과 필터 시스템의 설정을 구성합니다. 점막 표면의 혈관 패턴을 평가하기 위해 즉시 형광 염료의 정맥 내 주사 한 후 (예를 들면, FITC) 비특이적 혈액량 추적기위한 형광 내시경 검사를 수행한다. </li><li> 배경 비율에 더 나은 목표를 제공하기 위해 타겟 추적자 또는 &#39;스마트 프로브&#39;가지 경우에 추적 신청 후 몇 시간 이미징 고려하십시오. </li><li> 결과의 카메라, 비디오 문서를 수행합니다. </li></ol></li></ol> (5) 후 대장 내시경 검사 <ol><li> 빈 새장에 마우스를 분리하고 쓰레기를 흡입에서 마우스를 보호하기 위해 종이 타월에 누워. 저체온증 않도록 redlight 램프 마우스 따뜻한. 마우스를 준수하고 충분한 회복 될 때까지 방치하지 않는다의식은 흉골 드러 누움을 유지합니다. 완전히 의식이되면, 다시 각각의 케이지에 마우스를 놓습니다. </li><li> 그리고 작지만 누액 방지 상자 실험 도시 마우스의 끝에하면 CO 2 (100 % (v / v) 100 부피 %, 3 L / 분) 투여. 마우스가 완료 의식을 잃고 호흡이 정지 될 때까지 기다립니다. 경부 골절에 의한 파견 마우스입니다. 복부 개복술을 수행하고 콜론 이식편. 길이 방향으로 결장을 열고 더 조직 학적 또는 분자 평가를 씻는다. </li></ol>

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endomicroscopic+Imaging+of+COX-2+Activity+in+Murine+Sporadic+and+Colitis-Associated+Colorectal+Cancer.">Endomicroscopic Imaging of COX-2 Activity in Murine Sporadic and Colitis-Associated Colorectal Cancer.</a> Diagnostic and therapeutic endoscopy. 2013, 250641 (2013).</li><li>Bremer, C., Ntziachristos, V., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical-based+molecular+imaging:+contrast+agents+and+potential+medical+applications.">Optical-based molecular imaging: contrast agents and potential medical applications.</a> European radiology. 13, 231-243 (2003).</li><li>Keller, R., Winde, G., Terpe, H. J., Foerster, E. 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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+factor+from+a+transformed+cell+line+that+affects+cell+migration.">A factor from a transformed cell line that affects cell migration.</a> Proceedings of the National Academy of Sciences of the United States of America. 70, 369-372 (1973).</li><li>Msaki, 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=The+role+of+RelA+(p65)+threonine+505+phosphorylation+in+the+regulation+of+cell+growth,+survival,+and+migration.">The role of RelA (p65) threonine 505 phosphorylation in the regulation of cell growth, survival, and migration.</a> Molecular biology of the cell. 22, 3032-3040 (2011).</li><li>Zigmond, 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=Utilization+of+murine+colonoscopy+for+orthotopic+implantation+of+colorectal+cancer.">Utilization of murine colonoscopy for orthotopic implantation of colorectal cancer.</a> PloS one. 6, e28858 (2011).</li><li>Foersch, S., 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=Molecular+imaging+of+VEGF+in+gastrointestinal+cancer+in+vivo+using+confocal+laser+endomicroscopy.">Molecular imaging of VEGF in gastrointestinal cancer in vivo using confocal laser endomicroscopy.</a> Gut. 59, 1046-1055 (2010).</li><li>Mitsunaga, 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=Fluorescence+endoscopic+detection+of+murine+colitis-associated+colon+cancer+by+topically+applied+enzymatically+rapid-activatable+probe.">Fluorescence endoscopic detection of murine colitis-associated colon cancer by topically applied enzymatically rapid-activatable probe.</a> Gut. 62, 1179-1186 (2013).</li></ol>

저자가 공개하는 게 없다.

상피 상처 치유는 계속적인 과정이라고 할 수 있습니다. 위장 점막 내의 표면 세포의 연속 생리 각질은 상피 세포 16의 빈번한 재생을 필요로 발생합니다. 따라서, 장애인 상처 치유 위장 궤양과 17 문 합부 누출 (18) 등 여러 가지 질병에 엄청난 영향을 미친다. 상피를 자극하는 분자 배경의 평가뿐만 아니라 잠재적 인 약물 후보는 불완전 19,20 체외에서의 세?...

동물 모델은 크게 여러 장 병리에 대한 우리의 이해를 풍성하게했다. 실험실 마우스 (뮤스 musculus)는 인해 풍부한 유전자 및 게놈 정보를 생물 의학 연구에서 주요 동물 모델로 부상하고 유전자 녹아웃 균주에서 쉽게 사용할 수있다. 이해 질병 병인을 향상시키는 것 외에, 동물 모델은 또한 중요한 약물 후보뿐만 아니라 임상 진단이나 치료 개입을 테스트하기 위해 사용된다. 그러나, 인간의 질병을 흉내 낸 마우스 모델의 다양성에도 불구하고, 정기적으로 환자 치료에 사용되는 여러 진단 및 중재 적 옵션은 마우스 사용할 수 없습니다. 따라서, 감시 전략은 종종 간접 관찰에 한정되는 뮤린 질병 또는 치료 개입의 영향의 과정을 모니터링하거나 부검 분석 씀. 비 침습적 모니터링 마우스에 대한 질병 활동 지수와 같은 활력을 존재하지만, 한때체중 감소 또는 증가, 혈액, 소변 및 대변의 antification 다음은 간접 지표 간 약간의 차이에 의해 바이어스된다, 분석한다. 또한, 사후 부검은 반복 시점에서 종 관찰을 방지 분석합니다. 정교한 영상 기술은 최근 1,2 도입 된 마우스에서 질병 활동을 모니터링 할 수 있습니다. 이 영상 기술은 반복적 인 분석이 가능하지만, 그들은 단지 직접 점막의 시각화를 활성화 또는 생검 수집 또는 약물 후보의 국소​​ 및 점막 내암을 응용 프로그램으로 진단 또는 치료 개입을 허용하지 않는, 장에 설명하고 종종 부정확 한보기를 제공합니다. 최근 살아있는 마우스에 사용하기위한 고해상도의 내시경 시스템은 3,4 개발되었다. 처음 이러한 내시경 기술은 상처 치유 또는 장내 염증 제공 OBJ와 같은 endoluminal 대장 질환의 병리의 직접 시각화를 허용반복적 인 시점에서 동일 종의 동물 연구를 허용, 감정적, 실시간 상태. 이외에도 개별 마우스에서 반복 생검을 허용함으로써, 내시경 시스템은 또한 치료 적으로 관심 영역에 대한 물질의 직접적인 적용을 허용함으로써 구별 종양 또는 국소 염증에 영향을 미칠 수있다. 치료 및 제어 물질은 관심 영역에 직접 전달 될 수있다 더욱이,이 문제는 개인간 변동 제외한 동일한 마우스에서 수행 될 수있다. 이러한 시스템은 현재 이러한 대장염 심각도 (MEICS) 5-7의 뮤린 내시경 인덱스로 결장 염증, 상처 치유, 복강경 간 생검, 각종 점수 시스템을 사용하여 간 종양 8과 종양 발생의 동 소성 유도의 평가를 위해 사용되어왔다. 대장 벽의 비후, 혈관 패턴의 변화, 섬유소의 존재 mucos의 단위 : MEICS 염증을 평가하는 5 개의 매개 변수 구성알 표면 및 대변의 일관성. 이 프로토콜에서는 장내 상처 치유, 염증 및 결장암의 뮤린 모델에서 경질 내시경의 사용을 설명한다. 먼저, 상처 치유 및 염증의 대장 내시경 평가뿐만 아니라 대장염 활성 종 평가 및 뮤린 결장 cancerogenesis의 연구를 설명한다. 쥐 내시경의 사용 설명을 넘어, 우리는 조직 검사를 얻기 위해 내시경 장비의 사용에 대한 자세한 지침을 제공하고, 서로 다른 관심의 구성 요소 (예를 들어, 약물 후보 또는 종양 세포)의 국소 및 점막 내암 응용 프로그램입니다. 마지막으로, 우리는 대장 종양의 설정에서, 복잡한 분자 이미징 기술을 채용 뮤린 형광 내시경의 사용을 보여준다.

<table border="0" cellpadding="0" cellspacing="0" width="1467">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Name</td>
			<td style="width:423px;">Company</td>
			<td style="width:320px;">Catalogue Number</td>
			<td style="width:255px;">Comment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;"></td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Reagents</td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Alfalfa-free diet</td>
			<td>Harlan Laboritories, Madison, USA</td>
			<td style="width:320px;">2014</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Azoxymethane (AOM)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">A5486</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bepanthen eye ointment</td>
			<td style="width:423px;">Bayer, Leverkusen, Germany</td>
			<td style="width:320px;">80469764</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dextran sulphate sodium (DSS)</td>
			<td>TdB Consulatancy, Uppsala, Sweden</td>
			<td>DB001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Eosin</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 4382</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Ethylenediaminetetraacetic acid (EDTA)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 9884</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Falcon Tube 50ml</td>
			<td>BD Biosciences, Erembodegem, Belgium</td>
			<td style="width:320px;">352070</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Florene 100V/V</td>
			<td>Abbott, Wiesbaden, Germany</td>
			<td style="width:320px;">B506</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein-Isothiocyanat (FITC)&#8211;dextrane&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">FD4-250MG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Haematoxylin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">HHS32-1L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Isopentane (2- Methylbutane)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">M32631-1L</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Methylene blue</td>
			<td style="width:423px;">Merck, Darmstadt, Germany</td>
			<td style="width:320px;">1159430025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">O.C.T. Tissue Tek compound&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sakura, Zoeterwonde, Netherlands</td>
			<td style="width:320px;">4583</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Omnican F - canula</td>
			<td style="width:423px;">Braun, Melsungen, Germany</td>
			<td style="width:320px;">9161502</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Phosphate buffered saline, PBS</td>
			<td style="width:423px;">Lonza, Verviers, Belgium</td>
			<td style="width:320px;">4629</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium Chloride 0,9%</td>
			<td>Braun, Melsungen, Germany</td>
			<td style="width:320px;">5/12211095/0411</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Standard diet</td>
			<td>Altromin, Lage, Germany</td>
			<td style="width:320px;">1320</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tissue-Tek Cryomold</td>
			<td>Sakura, Leiden, Netherlands</td>
			<td style="width:320px;">4566</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Vitro &#8211; Clud&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">R. Langenbrinck, Teningen, Germany</td>
			<td style="width:320px;">04-0002&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">AIDA Control</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 096020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 716/40</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 809/81</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Biopsy Forceps, 3 Fr., 28cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61071ZJ</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dell Monitor</td>
			<td>Dell, Frankfurt am Main, Germany</td>
			<td style="width:320px;">U2412Mb</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029D</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029C</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fiber Optic Light Cable, 3.5mm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69495NL</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein Blue Filter System</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100032</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fluorescein Barrier Filter</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100033</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Foot switch</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20010430</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">HOPKINS Telescope, 1.9mm, Length 10cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">1830231</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB D-light P&#160;</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 133720</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB tricam SL II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 2230 20</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tubing set instruments VETPUMP II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69811</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tricam PDD PAL</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20221037</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">UniVet Porta</td>
			<td>Groppler Medizintechnik, Deggendorf, Germany</td>
			<td style="width:320px;">BKGM 0451</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Vetpump 2</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69321620</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine endoscopy for in vivo multimodal imaging of carcinogenesis and assessment of intestinal wound healing and inflammation

마우스 모델은 널리 인간 질병의 발병 기전을 연구 및 전임상 진단 절차뿐만 아니라 치료 적 개입을 평가하는 데 사용됩니다. 그러나, 병리학 적 변화의 올바른 평가는 종종 조직 학적 분석을 필요로하고, 생체 외에서 수행 할 때, 동물의 사망을 필요로한다. 따라서 기존의 실험 설정에서, 내 개별 추적 검사는 거의 가능하다. 따라서, 살아있는 마우스에서 뮤린 내시경의 개발은 직접 위장 점막을 시각화와 같은 변경을 모니터링하는 절차를 반복 모두 처음 조사자 수있다. 생체 쥐 내시경 검사에 대한 수많은 응용 프로그램, 장 염증이나 상처 치유를 공부 반복 점막 생검 획득을 포함한, 존재하고,에 로컬 소형 주입 카테터를 사용하여 진단 또는 치료 에이전트를 관리 할 수 있습니다. 가장 최근에, 분자 영상은 확장했다 영상 진단 모특정 photoprobes를 구별하여 표적 분자의 검출을 허용하는 특정 dalities. 결론적으로, 쥐의 내시경 검사는 생체 영상 진단 실험을위한 새로운 첨단 기술로 떠오르고있다 상당히 다양한 분야에서 임상 연구에 영향을 미칠 수 있습니다.

장 상처 치유의 생체 모니터링에서 (;도 1a 1mm 같음) 루틴 내시경 동안 점막 상처 3 프랑스어의 직경 미니어처 생검 겸자에 의해 기계적으로 유도 하였다. 이어서, 상처 치유를 매일 내시경 검사에 의해 모니터링 및 예 이미지 편집 소프트웨어를 사용하여 잔류 상처 면적의 측정에 의해 정량 하였다, ImageJ에 (도 1b). 시간에 따른 개개의 ...

Watch this Scientific Journal Video about Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation at JoVE.com

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

에 대한 쥐의 내시경<em&gt; 생체</em&gt; 발암 및 소장의 평가의 멀티 모달 이미징 상처 치유 및 염증

작은 동물 이미징 기술은 직렬 진단 시험 및 생체 내에서 치료 적 개입을 허용한다. 최근, 응용 프로그램의 범위가 크게 확대하고 현재 대장 종양 발생의 평가, 상처 치유 및 염증의 모니터링이 포함되어 있습니다. 이 프로토콜은 쥐 내시경 검사의 이러한 다양한 응용 가능성을 보여줍니다.

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

murine-endoscopy-for-vivo-multimodal-imaging-carcinogenesis

Research

JoVE Journal

Medicine

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

18.5K 조회수.  University Hospital Münster.  작은 동물 이미징 기술은 직렬 진단 시험 및 생체 내에서 치료 적 개입을 허용한다. 최근, 응용 프로그램의 범위가 크게 확대하고 현재 대장 종양 발생의 평가, 상처 치유 및 염증의 모니터링이 포함되어 있습니다. 이 프로토콜은 쥐 내시경 검사의 이러한 다양한 응용 가능성을 보여줍니다. 

비디오: Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

염증성 플라크의 적외선 형광 (NIRF) Intravascular 분자 이미징, Multimodal 접근 가까운 생체내에서 죽상 경화증의 영상을

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

연구

의학

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the complex cellular and molecular of intestinal pathology. Although the tunica mucosa is often the organ of interest in many inflammatory diseases, recent works demonstrated that the muscularis externa (ME) is also a highly immunocompetent organ that harbours a dense network of resident immunocytes.1,2 These works were performed within the standardized model of intestinal manipulation (IM) that leads to inflammation of the bowel wall, mainly limited to the ME. Clinically this inflammation leads to prolonged intestinal dysmotility, known as postoperative ileus (POI) which is a frequent and unavoidable complication after abdominal surgery.3 The inflammation is characterized by liberation of proinflammatory mediators such as IL-64 or IL-1&beta; or inhibitory neurotransmitters like nitric oxide (NO).5 Subsequently, tremendous numbers of immunocytes extravasate into the ME, dominated by polymorphonuclear neutrophils (PMN) and monocytes and finally maintain POI.2 Lasting for days, this intestinal paralysis leads to an increased risk of aspiration, bacterial translocation and infectious complications up to sepsis and multi organ failure and causes a high economic burden.6
In this manuscript we demonstrate the standardized model of IM and in vivo assessment of gastrointestinal transit (GIT) and colonic transit. Furthermore we demonstrate a method for separation of the ME from the tunica mucosa followed by immunological analysis, which is crucial to distinguish between the inflammatory responses in these both highly immunoactive bowel wall compartments. All analyses are easily transferable to any other research models, affecting gastrointestinal function.

Postoperative ileus (POI) is a complication of abdominal surgery leading to increased morbidity and a prolonged hospital stay. Because prophylactic or therapeutic strategies are lacking intensified research is necessary. Therefore we established a standardized and feasible mouse model to investigate the pathophysiology of POI and to study potential therapeutic options.

장 조작의 표준화 된 모델의 분석 : 쥐의 장 운동성 및 내장 벽의 염증의 기능 평가

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

상처 치유의 생쥐 모델

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

돌이킬 수 및 가역 일방적 Ureteric 폐색의 쥐 모델

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

대동맥 석회화와 염증의 혈관 평활근 세포 및 이미지의 석회화

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

Multimodal Quantitative Phase Imaging with Digital Holographic Microscopy Accurately Assesses Intestinal Inflammation and Epithelial Wound Healing

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The etiology of IBD remains unknown and current therapeutic strategies are based on the unspecific suppression of the immune system. The development of treatments that specifically target intestinal inflammation and epithelial wound healing could significantly improve management of IBD, however this requires accurate detection of inflammatory changes. Currently, potential drug candidates are usually evaluated using animal models in vivo or with cell culture based techniques in vitro. Histological examination usually requires the cells or tissues of interest to be stained, which may alter the sample characteristics and furthermore, the interpretation of findings can vary by investigator expertise. Digital holographic microscopy (DHM), based on the detection of optical path length delay, allows stain-free quantitative phase contrast imaging. This allows the results to be directly correlated with absolute biophysical parameters. We demonstrate how measurement of changes in tissue density with DHM, based on refractive index measurement, can quantify inflammatory alterations, without staining, in different layers of colonic tissue specimens from mice and humans with colitis. Additionally, we demonstrate continuous multimodal label-free monitoring of epithelial wound healing in vitro, possible using DHM through the simple automated determination of the wounded area and simultaneous determination of morphological parameters such as dry mass and layer thickness of migrating cells. In conclusion, DHM represents a valuable, novel and quantitative tool for the assessment of intestinal inflammation with absolute values for parameters possible, simplified quantification of epithelial wound healing in vitro and therefore has high potential for translational diagnostic use.

Accurate assessment of anti-inflammatory effects is of utmost importance for the evaluation of potential new drugs for the treatment of inflammatory bowel disease. Digital holographic microscopy provides assessment of inflammation in murine and human colonic tissue samples as well as automated multimodal evaluation of epithelial wound healing in vitro.

디지털 홀로그램 현미경과 복합 양적 위상 이미징 정확하게 장내 염증과 상피 상처 치유를 평가

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The ...

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not routinely employed in animal models. Conventional experimental readouts often rely on post mortem and ex vivo analyses, which prevent intra-individual follow-up examinations and increase the number of study animals needed. Fluorescence-mediated tomography enables the non-invasive, repetitive, quantitative, three-dimensional assessment of fluorescent probes. It is highly sensitive and permits the use of molecular makers, which allows for the specific detection and characterization of distinct molecular targets. In particular, targeted probes represent an innovative tool for analyzing gene activation and protein expression in inflammation, autoimmune disease, infection, vascular disease, cell migration, tumorigenesis, etc. In this article, we provide step-by-step instructions on this sophisticated imaging technology for the in vivo detection and characterization of inflammation (i.e., F4/80-positive macrophage infiltration) in a widely used murine model of intestinal inflammation. This technique might also be used in other research areas, such as immune cell or stem cell tracking.

Target-specific probes represent an innovative tool for analyzing molecular mechanisms, such as protein expression in various types of disease (e.g., inflammation, infection, and tumorigenesis). In this study, we describe a quantitative three-dimensional tomographic assessment of intestinal macrophage infiltration in a murine model of colitis using F4/80-specific fluorescence-mediated tomography.

검색 및 대 식 세포 관련 Murine 창 자 염증의 정량화에 대 한 단층 형광 중재

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During In Vitro Wound Healing

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated experimental procedures, while potentially lacking in precision. In vitro culture of epithelial cell lines provides a suitable platform for researching the cell migratory behavior in wound healing and the impact of treatments on these cells. The physiology of epithelial cells is often studied in non-confluent conditions; however, this approach may not resemble natural wound healing conditions. Disrupting the epithelium integrity by mechanical means generates a realistic model, but may impede the application of molecular techniques. Consequently, microscopy based techniques are optimal for studying epithelial cell migration in vitro. Here we detail two specific methods, the artificial wound scratch assay and the artificial migration front assay, that can obtain quantitative and qualitative data, respectively, on the migratory performance of epithelial cells.

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During  In Vitro Wound Healing

This manuscript describes how incision-like lesions made on cultured epithelial cell monolayers conveniently model wound healing in vitro, allowing for imaging by confocal or laser scanning microscopy, and which can provide high-quality quantitative and qualitative data for studying both cell behavior and the mechanisms involved in migration.

현미경 기반 방법을 생체 외에서 상처 치유 중 상피 세포 이동의 평가 대 한

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

각 막에 대 한 모델로 눈 버와 각 막 상피 마모 상처 치유

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two dimensional (2D) and three dimensional (3D) culture has generated a wealth of information not only about corneal biology but also about wound healing, myofibroblast biology, and scarring in general. The goal of the protocol is an assay system for quantifying myofibroblast development, which characterizes scarring. We demonstrate a corneal organ culture ex vivo model using pig eyes. In this anterior keratectomy wound, corneas still in the globe are wounded with a circular blade called a trephine. A plug of approximately 1/3 of the anterior cornea is removed including the epithelium, the basement membrane, and the anterior part of the stroma. After wounding, corneas are cut from the globe, mounted on a collagen/agar base, and cultured for two weeks in supplemented-serum free medium with stabilized vitamin C to augment cell proliferation and extracellular matrix secretion by resident fibroblasts. Activation of myofibroblasts in the anterior stroma is evident in the healed cornea. This model can be used to assay wound closure, the development of myofibroblasts and fibrotic markers, and for toxicology studies. In addition, the effects of small molecule inhibitors as well as lipid-mediated siRNA transfection for gene knockdown can be tested in this system.

A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment.

에 대한 쥐의 내시경

요약

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