Name: using digital image correlation to characterize local strains on vascular tissue specimens
Uploaded: 2016-01-24T14:00:00.000+00:00
Duration: 9 min 29 s
Description: Watch this Scientific Journal Video about Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens at JoVE.com

소프트웨어 및 기술 지원은 상관 솔루션 통합 (www.correlatedsolutions.com)의 호의였다.

참고 : 아래에서 설명하는 절차는 컬럼비아 사우스 캐롤라이나 대학, 사우스 캐롤라이나에서 기관 동물 케어 및 사용위원회에 의해 승인 된 프로토콜의 일환으로 수행되었다. 1. 조직 수집 및 해부 <ol><li> 조직 해부하기 전에 모든 수술 도구를 소독. 오토 클레이브 외과 가위 및 정밀 표준 집게뿐만 아니라 15 분 15 PSI의 압력과 121 ° C의 온도에서 수술 블레이드. </li><li> 신선한 돼지 (7 개월 된 랜드 레이스 남성, 60-70kg) 지역 도축장에서 그대로 대동맥과 신장 한 세트를 획득. 다시 아이스 1 % 인산염 완충 식염수 (PBS) 용액에 실험실에 전송 티슈. </li><li> 즉시 도착시, 수술 가위와 집게를 사용하여 주변 조직에서 복부 대동맥을 격리 할 것. </li><li> PBS (PH 7.2)로 가득 50 ML의 주사기를 사용하여 용기를 세 번 씻으십시오. 가위와 집게를 사용하면, 포스만큼 혈관 주위 조직을 제거시료의 완전성을 손상시키지 않고 환되는. </li><li> 세로 용기의 중간 부 상에 날카로운 면도날을 배치하고 용기 길이 방향 축에 수직 인 확인. 면도날 세 순차적 원주 인하를 적용하여 약 20 mm의 폭과 두 개의 링 샘플 각을 만듭니다. </li><li> 수직 블레이드 반경 방향으로 배향되도록, 하나의 링 시료에 날카로운 면도날을 위치. 단축 기계 테스트 스트립 형태의 샘플 결과 반경 컷을 산출하기 위해 급성 힘을 적용합니다. 100mm 유리 페트리 접시에 샘플을 놓고 표면의 반점의 응용 프로그램까지 PBS에 잠수함. 제 2 링 샘플에 대해 반복합니다. </li></ol> 표면 스페 클 패턴 2. 창조 <ol><li> 압력 밸브에 에어 브러쉬를 연결합니다. </li><li> 노즐 직경의 노즐 60 ~ 100 μm의의 얼룩을 산출 할 수있는 에어 브러쉬의 직경 (적절한 범위를 조정해야) 예비 연구로부터 결정될. </li><li> 에어 브러쉬의 중력 공급기에 검은 조직 마킹 염료의 약 2 ml에 붓고. </li><li> 약 0.5 미터 떨어진 샘플에서 에어 브러쉬를 놓습니다. </li><li> 페트리 접시에서 샘플을 제거합니다. PSI (100)의 분사 압력하에 약 5 초 동안 샘플의 내막 조직 표면에 마킹 염료 스프레이. 스페 클 패턴이 균일하게 시료 표면을 커버하도록 세 번 반복합니다. </li></ol> 실험 3. 성능 <ol><li> 조직 접착제를 사용하여 플라스틱 스트립 (1cm 폭 X 1cm 길이 X 0.5 cm 두께)에 샘플의 양 끝을 연결합니다. 조직 커팅 보드에 샘플을 놓습니다. 이 평평하도록 샘플을 놓고 디지털 캘리퍼스를 사용하여 치수를 측정한다. </li><li> 기계적 시험에 대한 시스템 제어를 시작합니다. 시스템이 홈 화면을 제어에서 &quot;설정&quot;탭에있는 작업 표시 줄의 &quot;파형&quot;를 선택합니다. </li><li> 광고-4 mm (시스템에 지정된 초기 위치에 대하여 4mm 연장)에 기계적 시험기의 상부 그립 바로 위치. 조심스럽게 기계 테스터의 상부 그립에 한 플라스틱 스트립 (3.1 시편에 부착) 확보 및 샘플 자유롭게 중지 할 수 있습니다. 샘플 및 상부 그립 사이의 거리가 2mm 미만인되도록 디지털 캘리퍼를 사용한다. </li><li> 샘플의 자유 단을 연장하지 않고 확보 할 수 있도록 수동 하부 그립의 위치를​​ 조정한다. 부드럽게 기계적 시험기의 하부 그립으로 샘플의 자유 단에 부착 된 플라스틱 스트립을 고정. <ol><li> 샘플 및 하부 그립 사이의 거리가 2mm 미만인되도록 디지털 캘리퍼를 사용한다. 시스템 로드셀 제로. 시료의 길이를 측정하고 글로벌 원주 균주의 계산을위한 기준 길이로 사용. </li></ol></li><li> 기계적 시험 프로토콜을 입력합니다. t에 사용되는 프로토콜그의 데모 0.01 mm / sec의 속도로 변위 18 % 시료 길이를 확장 4 축 변위 사이클을 수반한다. </li><li> 일시적으로는 수화 유지하기 위해 나머지 시험 프로토콜을 통해 샘플에 PBS 스프레이. </li><li> 로딩 프레임으로부터 1.5 m 위치된다 삼각대에 카메라 (5 메가 픽셀 카메라, 100mm 렌즈, 3.49 ㎛의 화소 크기) 마운트. 카메라와 시료 표면 깊이의 가장 낮은 필드에 카메라를 설정하고보기의 전체 필드에 포커스가되도록의 정렬을 조작하여 수직 확인합니다. </li><li> 촬상 소프트웨어를 연다. </li><li> 옵션 &quot;을 선택 시스템&quot;에서 &quot;PGR-2&quot;를 선택합니다. </li><li> 이미지를 저장할 수있는 프로젝트 경로를 선택 분석합니다. </li><li> &quot;타임 스퀘어&quot;아이콘을 클릭하고 5 초로 획득 간격을 지정합니다. </li><li> 샘플의 명확도를 얻을 수 있도록 렌즈의 노출, 개구 수 및 초점을 조절. </li><li> 샘플에 적절한 조명을 제공하는 LED의 위치를​​ 조정합니다. </li><li> 시료 표면의 화상을 얻을 촬상 소프트웨어에서 &quot;시작&quot;아이콘을 클릭. </li><li> 이미지 분석 소프트웨어를 엽니 다. </li><li> 획득 된 이미지를 가져옵니다. 개인 반점을 확대 한 다음이 개인 반점에서 픽셀 수를 계산합니다. 참고 : 대표 검은 반점을 확인합니다. 유사한 높은 값을 갖는 스페 클 양쪽 픽셀 사이의 직선 거리로 스펙 클 크기를 정의. 허용되는 스페 클의 크기, 전형적인 스펙 클의 폭 방향의 픽셀 수는 3 픽셀보다 커야한다. 가능하면 측정의 공간 해상도를 향상시키기 위해, 대부분의 얼룩은 스펙 클의 폭을 가로 질러 더 이상 5-7 화소를 가져야한다. 따라서,이 경우의 일반적인 반점 사이 10 μm의 작은에서 23 μm의 선형 치수에서 가장 큰에서 범위 것입니다. 적절한 부분 집합을 확인하려면크기, 일반적인 서브 세트는 그 폭에 걸쳐 적어도 흰색 3과 3 검은 얼룩을 가져야한다. 전형적인 스펙 클 선형 치수 5 픽셀이면, 각 서브 세트는 선형 31x31 사이즈가 적어도 105 μm의 것이다. 서브 세트의 중심 간 간격은 선형 치수의 1/6 이상이어야한다. 따라서 31x31 서브 세트 크기, 거리는 직선 거리로 18 μm의 픽셀을 나타내는 5이다. </li><li> 스페 클 패턴의 품질을 확인한 후, 동시에 테스트를 시작하는 이미지 캡처 소프트웨어에서 시스템의 &quot;실행&quot;아이콘 &quot;시작&quot;아이콘을 클릭합니다. </li><li> 카메라와 촬상 소프트웨어를 사용하여 시험에 걸쳐 일련의 이미지를 캡쳐. </li></ol> 실험 후 4. 정리 절차 <ol><li> 생물 학적 가방에 폐기 샘플을 놓고 가방을 닫습니다. 적절한 처리를 위해 사우스 캐롤라이나 대학의 환경 보건 및 안전 (EHS)의 부서를 호출합니다. </li><li> 인산염을 준비증류수에 세제 살균제의 1:64 희석 비율을 무료로 소독액. 20 분 동안이 솔루션에 수술 도구를 만끽 해보세요. </li><li> 충분히 증류수 4.2에 설명 된 항목을 헹군다. 종이 타월을 이용하여 툴을 건조 후 70 % 에탄올 용액으로 스프레이 그들. 다시 종이 타월을 사용하여 수술 도구를 건조 및 수술 도구 상자에 다시 넣어. </li></ol> 로컬 스트레인 필드를 측정 5. 이미지 분석 <ol><li> 이미지 분석 소프트웨어를 엽니 다. </li><li> 분석 할 필요가있는 모든 이미지를 선택, &quot;얼룩 이미지&quot;탭을 클릭합니다. </li><li> 사각형 도구를 클릭하고 첫 번째 이미지에 대한 관심의 영역을 선택합니다. </li><li> 일부 크기 41 × 41 픽셀과 스텝 크기 5 픽셀 입력합니다. </li><li> 소프트웨어에서 분석 탭을 시작을 클릭 최적화 된 8 트랩으로 보간을 선택; 가우스와 같은 옵션 제로 정규화 제곱 차이와 일부의 무게와 같은 기준을 선택합니다. </li><li>소프트웨어의 기본값으로 임계 값 옵션을 설정합니다. </li><li> 시작 분석 탭의 후 처리 하위 탭을 클릭합니다. 옵션 변형 계산을 클릭하고 필터의 크기와 소프트웨어에 기본적으로 필터의 유형을 둡니다. 선택 텐서는 라그랑주을 입력합니다. </li><li> 데이터 탭을 선택하고 표면 변형 필드의 시각화에 대한 모든 분석 이미지를 선택합니다. </li></ol>

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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=Strain+field+measurements+on+mouse+carotid+arteries+using+microscopic+three-dimensional+digital+image+correlation.">Strain field measurements on mouse carotid arteries using microscopic three-dimensional digital image correlation.</a> J Biomed Mater Res A. 84, 178-190 (2008).</li><li>Sutton, M. A., Orteu, J. J., Schreier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Image correlation for shape, motion and deformation measurements: basic concepts, theory and applications. , (2009).</li><li>Verhulp, E., van Rietbergen, B., Huiskes, 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+three-dimensional+digital+image+correlation+technique+for+strain+measurements+in+microstructures.">A three-dimensional digital image correlation technique for strain measurements in microstructures.</a> J Biomech. 37, 1313-1320 (2004).</li><li>Wang, C. C., Deng, J. M., Ateshian, G. A., Hung, C. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+automated+approach+for+direct+measurement+of+two-dimensional+strain+distributions+within+articular+cartilage+under+unconfined+compression.">An automated approach for direct measurement of two-dimensional strain distributions within articular cartilage under unconfined compression.</a> Journal of Biomechanical Engineering. 124, 557-567 (2002).</li><li>Franck, C., Hong, S., Maskarinec, S., Tirrell, D., Ravichandran, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+full-field+measurements+of+large+deformations+in+soft+materials+using+confocal+microscopy+and+digital+volume+correlation.">Three-dimensional full-field measurements of large deformations in soft materials using confocal microscopy and digital volume correlation.</a> Exp Mech. 47, 427-438 (2007).</li><li>Garcia, 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=Experimental+study+and+constitutive+modelling+of+the+passive+mechanical+properties+of+the+porcine+carotid+artery+and+its+relation+to+histological+analysis:+Implications+in+animal+cardiovascular+device+trials.">Experimental study and constitutive modelling of the passive mechanical properties of the porcine carotid artery and its relation to histological analysis: Implications in animal cardiovascular device trials.</a> Med Eng Phys. 33, 665-676 (2011).</li><li>Miller, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+test+very+soft+biological+tissues+in+extension?.">How to test very soft biological tissues in extension?.</a> J Biomech. 34, 651-657 (2001).</li><li>Sutton, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Springer handbook of experimental solid mechanics. , 565-600 (2008).</li><li>Han, H. C., Fung, Y. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+strain+of+canine+and+porcine+aortas.">Longitudinal strain of canine and porcine aortas.</a> J Biomech. 28, 637-641 (1995).</li><li>Sokolis, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+passive+strain-energy+function+for+elastic+and+muscular+arteries:+correlation+of+material+parameters+with+histological+data.">A passive strain-energy function for elastic and muscular arteries: correlation of material parameters with histological data.</a> Med Biol Eng Comput. 48, 507-518 (2010).</li><li>Zhou, B., Wolf, L., Rachev, A., Shazly, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+structure-motivated+model+of+the+passive+mechanical+response+of+the+primary+porcine+renal+artery.">A structure-motivated model of the passive mechanical response of the primary porcine renal artery.</a> J Mech Med Biol. , (2013).</li><li>Zhou, B., Rachev, A., Shazly, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+biaxial+active+mechanical+properties+of+the+porcine+primary+renal+artery.">The biaxial active mechanical properties of the porcine primary renal artery.</a> J Mech Behav Biomed Mater. 48, 28-37 (2015).</li><li>Sommer, G., Holzapfel, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+constitutive+modeling+of+the+biaxial+mechanical+response+of+intact+and+layer-dissected+human+carotid+arteries.">3D constitutive modeling of the biaxial mechanical response of intact and layer-dissected human carotid arteries.</a> J Mech Behav Biomed Mater. 5, 116-128 (2012).</li></ol>

저자는 관심의 잠재적 인 충돌이 없습니다.

이전의 연구는 샘플 18,20,21,23,24 균주를 평가 염료 비디오 추적 방법의 넓은 범위를 사용하였으나, 우리의 목표는 본 평가 용 2D-DIC와 커플 일축 인장 시험에 포괄적 인 방법을 제공하는 것이다 혈관 조직 샘플의 표면 변형. 샘플 축성 로딩이 일어나 고해상도 카메라와 실내 이미지 분석 소프트웨어를 변형 필드는 소정의 표면 영역 내에 측정 할 수있다. 혈관 조직의 기계적 시험에 특히 관?...

오십년 이상에 걸친 연구의 풍부한 역사는 혈관 조직의 기계적 특성을 정량화에 초점을 맞추고있다. 이러한 연구는 우리가 더 혈관 병리 생리 및 동작 모두 이해 효능 / 혈관 내 장치의 호환성 및 지원 설계 및 엔지니어링 혈관의 제조 1-6 구조를 평가하기위한 기초를 제공 할 수있다. 부드러운 조직과 기계적 특성의 구성적인 모델링의 기계적 응답의 정확한 측정은 본질적으로 대부분의 조직 유형에 의해 전시 된 기계적인 이질성, 이방성 및 비선형 성으로 인해 도전하고있다. 또한, 실험 측정은 종종 지역의 기계적 시험의 과정에서 샘플 그립 인터페이스를 도입 복잡성 (즉, 굴곡, 마찰, 응력 집중, 찢어) 및 조직은 살아있는 동물에서 적출되면 기계적 성질의 피할 수없는 변화에 의해 혼동된다. &lt;/ P&gt; 단축 인장 실험은 고체 재료로 이루어진 시험편에 대해 수행 할 수있는 간단한 기계적 시험 사이이며, 종종 혈관 조직의 기계적인 반응을 평가하는 데 사용된다. 이러한 실험 결과는 모두 천연 및 조직 공학 소스에 유용한 사전 정보를 제공하고, 혈관 벽 7-11의 기계적 동작에 대해 특정 치료, 질환 상태, 또는 화합물의 약물 학적 효과를 비교하는데 사용될 수있다. 연부 조직의 단축 기계적 시험은 일반적으로 가장 일반적으로 개 뼈 또는 링 7,8,12-14 모양의 비교적 균일 한 형상으로 샘플에서 수행됩니다. 그러나 이러한 이상화 기하학에서 중요한 출발 인해 테스트 시스템 내에서 조직의 절개, 격리 및 클램핑과 관련된 문제로 발생할 수 있습니다. 기하학의 모든 비 균일 성이 궁극적으로 이기종 스트레스와 긴장을 야기 할 것이다필드는 샘플의 실제 샘플 형상에 의존 이질성뿐만 아니라, 샘플 크기 (그립에 대하여) 및 9,15,16 재료의 기계적 성질과 정도, 축 확장을 실시하는 경우. 필드 불균질성이 중요 할 때, 상대적으로 파지 위치에 기초하여 샘플 스트레인 계산은 부정확 및 기계적 행동을 평가하기위한 기초 따라서 불충분하다. 영상 분석 시스템은 널리 종종 시편 표면 (17, 18)에인가 높은 조영제 마커를 이용하여 연부 조직의 변형률 측정 용으로 사용되어왔다. 디지털 이미지의 상관 관계, 변형 전후의 시험편의 표면에 계조 강도 값을 비교하여 전체 필드 표면 변형을 측정하는 광학 측정 학적 기법, 연조직 19-21 분석 비디오와 함께 사용되어왔다. 디지털 화상 상관의 여러 가지 이점 interferometr 비교있다측정을 위해 사용될 수있는 IC 방법. 우선, 비접촉 측정 기술로서, 이는 측정 시스템에 영향을 시험편되는 방식으로 인한 재료 특성을 수정 교란 효과를 최소화한다. 둘째, 덜 엄격한 측정 환경을 필요로하고 다른 방법에 비해 감도, 해상도의 넓은 범위를 갖는다. 셋째, 시야 전체의 캡처 기능 부여,이 기술은 평균과 지방 모두 기계적 응답을 특성화 할 수있다. 방법의 자세한 설명은 독자 서튼 (22)에 의해 책을 참조하는 것이 좋습니다. 시료 표면에 변형 필드를 획득하기 위해 2 차원 디지털 화상의 상관 기법 (2D-DIC)이 사용될 수있다. 즉, 시편의 이미지는 무부하 및 다양한로드 상태에서 캡처됩니다. 첫 번째 이미지는 후속 계산을위한 메쉬를 형성하는 부분 집합 (M × M 픽셀)라는 작은 사각형으로 나누어 져 있습니다2D 변형 필드. 변형 된 시료의 각 사각형의 위치가 이미지 매칭 알고리즘을 이용하여 획득된다. 각 정사각형의 동작은 다음의 다항식 피팅 또는 유한 요소 보간을 포함하는 다양한 방법을 통해 변형 구배 및 변형을 유도하는 데 사용할 수있는 변위 필드를 수득 화상별로 화상 추적된다. 본 논문에서는 단축 인장 시험 및 2D-DIC의 통합을 통해 네이티브 혈관 조직의 표면 변형 분야의 평가를위한 상세한 방법을 제공한다.

<table><tbody><tr><td>Uniaxial tensile mechanical tester</td><td>Enduratec</td><td>3230 AT/HR</td><td></td></tr><tr><td>Blue tissue marking dye</td><td>http://www.ebay.com/itm/Tissue-Marking-Dye-in-Bottles-2oz-Bottle-1-ea-/201193551510?pt=LH_DefaultDomain_0&amp;amp;hash=item2ed811f696</td><td></td><td></td></tr><tr><td>Sprayer</td><td>Anest-iwata</td><td>CM-B</td><td>Custom Micron B</td></tr><tr><td>Camera</td><td>Point Grey</td><td>GS2-GE-50S5M-C</td><td></td></tr><tr><td>Lens</td><td>Tokina</td><td>AT-X M100</td><td></td></tr><tr><td>Vascular tissue</td><td>Caughman Inc</td><td></td><td></td></tr><tr><td>0.9% Sodium Chloride Injection PBS</td><td>BAXTER HEALTHCARE CORP.</td><td></td><td></td></tr><tr><td>Vic_snap</td><td>Correlated Solutions</td><td></td><td></td></tr><tr><td>Vic_2D</td><td>Correlated Solutions</td><td></td><td></td></tr><tr><td>Wintest 4.1</td><td>Bose ElectroForce</td><td></td><td></td></tr><tr><td>Tissue adhesive</td><td>&amp;nbsp;3M Vetbond&amp;nbsp;</td><td>1469SB</td><td></td></tr><tr><td>Disinfectant&amp;nbsp;</td><td>Fisher Scientific</td><td>04-355-13</td><td>Decon BDD Bacdown Detergent Disinfectant</td></tr></tbody></table>

using digital image correlation to characterize local strains on vascular tissue specimens

Characterization of the mechanical behavior of biological and engineered soft tissues is a central component of fundamental biomedical research and product development. Stress-strain relationships are typically obtained from mechanical testing data to enable comparative assessment among samples and in some cases identification of constitutive mechanical properties. However, errors may be introduced through the use of average strain measures, as significant heterogeneity in the strain field may result from geometrical non-uniformity of the sample and stress concentrations induced by mounting/gripping of soft tissues within the test system. When strain field heterogeneity is significant, accurate assessment of the sample mechanical response requires measurement of local strains. This study demonstrates a novel biomechanical testing protocol for calculating local surface strains using a mechanical testing device coupled with a high resolution camera and a digital image correlation technique. A series of sample surface images are acquired and then analyzed to quantify the local surface strain of a vascular tissue specimen subjected to ramped uniaxial loading. This approach can improve accuracy in experimental vascular biomechanics and has potential for broader use among other native soft tissues, engineered soft tissues, and soft hydrogel/polymeric materials. In the video, we demonstrate how to set up the system components and perform a complete experiment on native vascular tissue.

혈관 조직에 경 사진 축 확장 시험에서 취득한 기계 데이터는 주어진 변위 속도로 적용 샘플 변위 관계 대 부하로 구성되어 있습니다. 본 연구에서는 축성 기계적 시험과 함께, 2D-DIC 다양한 변형 상태에서 직교하는 방향에서 시료의 표면 변형 필드를 측정하기 위해 사용된다. 혈관 조직의 점탄성 특성은 기계적 전처리 이전의 하중 - 변위 곡선 이력의 주목할만한 정도에 ?...

Watch this Scientific Journal Video about Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens at JoVE.com

Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens

디지털 이미지의 상관 관계를 사용하면 혈관 조직 표본의 로컬 균주의 특성을

We describe the use of digital image correlation to characterize the local surface strain field on vascular tissue samples subjected to uniaxial tensile testing. These measurements facilitate precise quantification of the sample mechanical response and the generation of constitutive stress-strain relations.

Characterization of the mechanical behavior of biological and engineered soft tissues is a central component of fundamental biomedical research and ...

using-digital-image-correlation-to-characterize-local-strains-on

Research

JoVE Journal

Biology

9.4K 조회수.  University of South Carolina. We describe the use of digital image correlation to characterize the local surface strain field on vascular tissue samples subjected to uniaxial tensile testing. These measurements facilitate precise quantification of the sample mechanical response and the generation of constitutive stress-strain relations.

비디오: Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens

Monitoring the Wall Mechanics During Stent Deployment in a Vessel

Clinical trials have reported different restenosis rates for various stent designs1. It is speculated that stent-induced strain concentrations on the arterial wall lead to tissue injury, which initiates restenosis2-7. This hypothesis needs further investigations including better quantifications of non-uniform strain distribution on the artery following stent implantation. A non-contact surface strain measurement method for the stented artery is presented in this work. ARAMIS stereo optical surface strain measurement system uses two optical high speed cameras to capture the motion of each reference point, and resolve three dimensional strains over the deforming surface8,9. As a mesh stent is deployed into a latex vessel with a random contrasting pattern sprayed or drawn on its outer surface, the surface strain is recorded at every instant of the deformation. The calculated strain distributions can then be used to understand the local lesion response, validate the computational models, and formulate hypotheses for further in vivo study.

Stent-induced arterial strain distributions are characterized using an optical surface strain measurement system. This visualization technique is used to gain insights into the impact of stent implantation on the host vessel.

선박의 스텐 트를 배포하는 동안 벽 역학 모니터링

Clinical trials have reported different restenosis rates for various stent designs1. It is speculated that stent-induced strain concentrations on the ...

연구

생체공학

Observing and Quantifying Fibroblast-mediated Fibrin Gel Compaction

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global reorganization and realignment of the gel microstructure. This process proceeds in a complex manner that is dependent in part on the interplay between the location of the cells, the geometry of the gel, and the mechanical constraints on the gel. To better understand how these variables produce global fiber alignment patterns, we use time-lapse differential interference contrast (DIC) microscopy coupled with an environmentally controlled bioreactor to observe the compaction process between geometrically spaced explants (clusters of fibroblasts). The images are then analyzed with a custom image processing algorithm to obtain maps of the strain. The information obtained from this technique can be used to probe the mechanobiology of various cell-matrix interactions, which has important implications for understanding processes in wound healing, disease development, and tissue engineering applications.

Time-lapse microscopy and image processing techniques were used to observe and analyze fibroblast-mediated gel compaction and fibrin fiber realignment in an environmentally controlled bioreactor over a 48 hr&#160;period.

섬유아세포 매개 피브린 겔 다짐 관찰 및 정량화

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global ...

Full-field Strain Measurements for Microstructurally Small Fatigue Crack Propagation Using Digital Image Correlation Method

A novel measurement approach is used to reveal the cumulative deformation field at a sub-grain level and to study the influence of microstructure on the growth of microstructurally small fatigue cracks. The proposed strain field analysis methodology is based on the use of a unique pattering technique with a characteristic speckle size of approximately 10 &#181;m. The developed methodology is applied to study the small fatigue crack behavior in body centered cubic (bcc) Fe-Cr ferritic stainless steel with a relatively large grain size allowing a high spatial measurement accuracy at the sub-grain level. This methodology allows the measurement of small fatigue crack growth retardation events and associated intermittent shear strain localization zones ahead of the crack tip. In addition, this can be correlated with the grain orientation and size. Thus, the developed methodology can provide a deeper fundamental understanding of the small fatigue crack growth behavior, required for the development of robust theoretical models for the small fatigue crack&#160;propagation in polycrystalline materials.

Microstructurally small fatigue crack growth behavior is investigated using a novel methodological approach combining crack growth rate measurement and strain-field analysis to reveal the cumulative deformation field at sub-grain level.

A novel measurement approach is used to reveal the cumulative deformation field at a sub-grain level and to study the influence of microstructure on the ...

공학

Intermediate Strain Rate Material Characterization with Digital Image Correlation

The mechanical response of a material under dynamic load is typically different than its behavior under static conditions; therefore, the common quasistatic equipment and procedures used for material characterization are not applicable for materials under dynamic loads. The dynamic response of a material depends on its deformation rate and is broadly categorized into high (i.e., greater than 200/s), intermediate (i.e., 10&#8722;200/s) and low strain rate regimes (i.e., below 10/s). Each of these regimes calls for specific facilities and testing protocols to ensure the reliability of the acquired data. Due to the limited access to high-speed servo-hydraulic facilities and validated testing protocols, there is a noticeable gap in the results at the intermediate strain rate. The current manuscript presents a validated protocol for the characterization of different materials at these intermediate strain rates. Strain gauge instrumentation and digital image correlation protocols are also included as complimentary modules to extract the utmost level of detailed data from every single test. Examples of raw data, obtained from a variety of materials and test setups (e.g., tensile and shear) is presented and the analysis procedure used to process the output data is described. Finally, the challenges of dynamic characterization using the current protocol, along with the limitations of the facility and methods of overcoming potential problems are discussed.

Here we present a methodology for the dynamic characterization of tensile specimens at intermediate strain rates using a high-speed servo-hydraulic load frame. Procedures for strain gauge instrumentation and analysis, as well as for digital image correlation strain measurements on the specimens, are also defined.

The mechanical response of a material under dynamic load is typically different than its behavior under static conditions; therefore, the common ...

A Method to Estimate Cadaveric Femur Cortical Strains During Fracture Testing Using Digital Image Correlation

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral surface obtained from mechanical testing. This optical method requires a texture of many contrasting fiduciary marks on a solid white background for accurate tracking of surface deformation as loading is applied to the specimen. Immediately prior to testing, the surface of interest in the camera view is painted with a water-based white primer and allowed to dry for several minutes. Then, a black paint is speckled carefully over the white background with special consideration for the even size and shape of the droplets. Illumination is carefully designed and set such that there is optimal contrast of these marks while minimizing reflections through the use of filters. Images were obtained through high speed video capture at up to 12,000 frames/s. The key images prior to and including the fracture event are extracted and deformations are estimated between successive frames in carefully sized interrogation windows over a specified region of interest. These deformations are then used to compute surface strain temporally during the fracture test. The strain data is very useful for identifying fracture initiation within the femur, and for eventual validation of proximal femur fracture strength models derived from Quantitative Computed Tomography-based Finite Element Analysis (QCT/FEA).

In this protocol, the femur surface strains are estimated during fracture testing using the digital image correlation technique. The novelty of the method involves application of a high-contrast stochastic speckle pattern on the femur surface, carefully specified illumination, high speed video capture, and digital image correlation analysis for strain calculations.

방법 골절 디지털 이미지 상관 관계를 사용 하 여 테스트 하는 동안 싱가포르로 대 퇴 골 피 질 긴장을 예상

This protocol describes the method using digital image correlation to estimate cortical strain from high speed video images of the cadaveric femoral ...

Measuring Local Tissue Strains in Tendons via Open-Source Digital Image Correlation

There is considerable scientific interest in understanding the strains that tendon cells experience in situ and how these strains influence tissue remodeling. Based on this interest, several analytical techniques have been developed to measure local tissue strains within tendon explants during loading. However, in several cases, the accuracy and sensitivity of these techniques have not been reported, and none of the algorithms are publicly available. This has made it difficult for the more widespread measurement of local tissue strains in tendon explants. Therefore, the objective of this paper was to create a validated analysis tool for measuring local tissue strains in tendon explants that is readily available and easy to use. Specifically, a publicly available augmented-Lagrangian digital image correlation (ALDIC) algorithm was adapted for measuring 2D strains by tracking the displacements of cell nuclei within mouse Achilles tendons under uniaxial tension. Additionally, the accuracy of the calculated strains was validated by analyzing digitally transformed images, as well as by comparing the strains with values determined from an independent technique (i.e., photobleached lines). Finally, a technique was incorporated into the algorithm to reconstruct the reference image using the calculated displacement field, which can be used to assess the accuracy of the algorithm in the absence of known strain values or a secondary measurement technique. The algorithm is capable of measuring strains up to 0.1 with an accuracy of 0.00015. The technique for comparing a reconstructed reference image with the actual reference image successfully identified samples that had erroneous data and indicated that, in samples with good data, approximately 85% of the displacement field was accurate. Finally, the strains measured in mouse Achilles tendons were consistent with the prior literature. Therefore, this algorithm is a highly useful and adaptable tool for accurately measuring local tissue strains in tendons.

Measuring Local Tissue Strains in Tendons via Open-Source Digital Image Correlation

This paper describes an open-source digital image correlation algorithm for measuring local 2D tissue strains within tendon explants. The accuracy of the technique has been validated using multiple techniques, and it is available for public use.

오픈 소스 디지털 이미지 상관관계를 통한 힘줄의 국소 조직 변형 측정

There is considerable scientific interest in understanding the strains that tendon cells experience in situ and how these strains influence tissue ...

Controlled Strain of 3D Hydrogels under Live Microscopy Imaging

External forces are an important factor in tissue formation, development, and maintenance. The effects of these forces are often studied using specialized in vitro stretching methods. Various available systems use 2D substrate-based stretchers, while the accessibility of 3D techniques to strain soft hydrogels, is more restricted. Here, we describe a method that allows external stretching of soft hydrogels from their circumference, using an elastic silicone strip as the sample carrier. The stretching system utilized in this protocol is constructed from 3D-printed parts and low-cost electronics, making it simple and easy to replicate in other labs. The experimental process begins with polymerizing thick (&#62;100 &#956;m) soft fibrin hydrogels (Elastic Modulus of ~100 Pa) in a cut-out at the center of a silicone strip. Silicone-gel constructs are then attached to the printed-stretching device and placed on the confocal microscope stage. Under live microscopy the stretching device is activated, and the gels are imaged at various stretch magnitudes. Image processing is then used to quantify the resulting gel deformations, demonstrating relatively homogenous strains and fiber alignment throughout the gel&#8217;s 3D thickness (Z-axis). Advantages of this method include the ability to strain extremely soft hydrogels in 3D while executing in situ microscopy, and the freedom to manipulate the geometry and size of the sample according to the user&#8217;s needs. Additionally, with proper adaptation, this method can be used to stretch other types of hydrogels (e.g., collagen, polyacrylamide or polyethylene glycol) and can allow for analysis of cells and tissue response to external forces under more biomimetic 3D conditions.

The presented method involves uniaxial stretching of 3D soft hydrogels embedded in silicone rubber while allowing live confocal microscopy. Characterization of the external and internal hydrogel strains as well as fiber alignment are demonstrated. The device and protocol developed can assess the response of cells to various strain regimes.

라이브 현미경 이미징 에서 3D 하이드로겔의 제어 변형

External forces are an important factor in tissue formation, development, and maintenance. The effects of these forces are often studied using specialized ...

A Method to Study the Correlation Between Local Collagen Structure and Mechanical Properties of Atherosclerotic Plaque Fibrous Tissue

The rupture of atherosclerotic plaques in coronary and carotid arteries is the primary cause of fatal cardiovascular events. However, the rupture mechanics of the heterogeneous, highly collagenous plaque tissue, and how this is related to the tissue's fibrous structure, are not known yet. Existing pipelines to study plaque mechanics are limited to obtaining only gross mechanical characteristics of the plaque tissue, based on the assumption of structural homogeneity of the tissue. However, fibrous plaque tissue is structurally heterogeneous, arguably mainly due to local variation in the collagen fiber architecture.
The mechano-imaging pipeline described here has been developed to study the heterogeneous structural and mechanical plaque properties. In this pipeline, the tissue's local collagen architecture is characterized using multiphoton microscopy (MPM) with second-harmonic generation (SHG), and the tissue's failure behavior is characterized under uniaxial tensile testing conditions using digital image correlation (DIC) analysis. This experimental pipeline enables correlation of the local predominant angle and dispersion of collagen fiber orientation, the rupture behavior, and the strain fingerprints of the fibrous plaque tissue. The obtained knowledge is key to better understand, predict, and prevent atherosclerotic plaque rupture events.

We have developed a mechano-imaging pipeline to study the heterogeneous structural and mechanical atherosclerotic plaque properties. This pipeline enables correlation of the local predominant angle and dispersion of collagen fiber orientation, the rupture behavior, and the strain fingerprints of the fibrous plaque tissue.

국소 콜라겐 구조와 죽상경화반 섬유조직의 기계적 특성 사이의 상관관계를 연구하는 방법

The rupture of atherosclerotic plaques in coronary and carotid arteries is the primary cause of fatal cardiovascular events. However, the rupture ...

A Novel Approach to Monitoring Graft Neovascularization in the Human Gingiva

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids direct contact with the measured area, it is suitable for monitoring blood flow changes during wound healing in human patients. Vestibuloplasty is periodontal surgery to the oral vestibule, aiming to restore vestibular depth with simultaneous enlargement of the keratinized gingiva. In this special clinical case, a split thickness flap was elevated at the first upper premolar and a xenogenic collagen matrix was adapted to the resulting recipient bed. LSCI was used to monitor the re- and neovascularization of the graft and the surrounding mucosa for one year. A protocol is introduced for the correct adjustment of microcirculation measurement in the oral mucosa, highlighting difficulties and possible failures.
The clinical case study presented demonstrated that &#8212; following the appropriate protocol &#8212; LSCI is a suitable and reliable method for following up microcirculation in a healing wound in the human oral mucosa and gives useful information on graft integration.

This study introduces a protocol for measuring microcirculation in human oral mucosa by laser speckle contrast imaging. The monitoring of wound healing after vestibuloplasty combined with a xenogenic collagen graft is presented on a clinical case.

새로운 접근 방식을 접목 Neovascularization 인간의 Gingiva에 모니터링

Laser speckle contrast imaging (LSCI) is a novel method for measuring superficial blood perfusion over large areas. Since it is non-invasive and avoids ...

의학

Quantification of Vascular Parameters in Whole Mount Retinas of Mice with Non-Proliferative and Proliferative Retinopathies

Retinopathies are a heterogeneous group of diseases that affect the neurosensory tissue of the eye. They are characterized by neurodegeneration, gliosis&#160;and a progressive change in vascular function and structure. Although the onset of the retinopathies is characterized by subtle disturbances in visual perception, the modifications in the vascular plexus are the first signs detected by clinicians. The absence or presence of neovascularization determines whether the retinopathy is classified as either non-proliferative (NPDR) or proliferative (PDR). In this sense, several animal models tried to mimic specific vascular features of each stage to determine the underlying mechanisms involved in endothelium alterations, neuronal death and other events taking place in the retina. In this article, we will provide a complete description of the procedures required for the measurement of retinal vascular parameters in adults and early birth mice at postnatal day (P)17. We will detail the protocols to carry out retinal vascular staining with Isolectin GSA-IB4 in whole mounts for later microscopic visualization. Key steps for image processing with Image J Fiji software are also provided, therefore, the readers will be able to measure vessel density, diameter and tortuosity, vascular branching, as well as avascular and neovascular areas. These tools are highly helpful to evaluate and quantify vascular alterations in both non-proliferative and proliferative retinopathies.

This article describes a well-established and reproducible lectin stain assay for the whole mount retinal preparations and the protocols required for the quantitative measurement of vascular parameters frequently altered in proliferative and non-proliferative retinopathies.