This project was supported by Grant Numbers HL96593 from NIH and D56HP20783 from HRSA/ HHS. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or HRSA / HHS.

주 : 모세관 이미지를 획득하기위한 획득 프로세스는 이전에 간행되었으며, 해당 이미지 획득 및 분석 컴퓨터 프로그램을 갖는 디지털 스틸 카메라를 이용하여 수행된다 (11) (21)이 실험실 분석을위한 영상 획득을 단순화 분석 아닌 동영상을 정지 영상을 이용한다.. 다음은 이미지에서 모세 혈관을 정량을위한 새로운 기술에 대해 설명합니다. 1. 이미지 향상 과정 <ol><li> 흑백 디지털 카메라와 디지털 이미지를 얻습니다. 이러한 카메라 눈금자로 알려진 길이의 물체의 사진을 촬영하여 알려진 크기의 객체 이미지들을 보정. 이 프로세스는 측정을 정확하게 처리 후 모세관을 계산하는 컴퓨터 프로그램을 허용한다. 이상적으로 높은 정밀도, 십자선 (그것으로 에칭 통치자와 유리의 과학적으로 제조 된 조각)이 사용되어야한다. 1mm 평방의 화소 수를 측정컴퓨터 프로그램을 사용하여 상자. 참고 :이 프로토콜의 재현성 및 표준화의 핵심은 계산됩니다 1mm 2 상자의 적절한 배치에 크게 의존한다. </li><li> 모세 혈관의 시각화를 극대화 배경, 모세 혈관을 어둡게하고 밝게 대비 향상 도구를 사용합니다. 배경에서 모세 혈관의 초기 분화 이후 단계에있는 이미지의 적절한 자르기 중요하다. 사용하여 이미지 조정하여이 작업을 수행에서이 클릭하려면 &quot;가장 적합한 히스토그램을.&quot;캡처, 강도, 이미지 히스토그램, 최적입니다. </li><li> 새로운 화상으로서 모세관 카운팅 ROI를 자르 (관심 영역을 선택). 1mm 대등 530 화소의 이미지를 보정하여 결정 하였다 1mm 2 상자를 사용한다. 자른 이미지는 이미지의 맨 위에 모세관 루프의 정점을 배치 있는지 확인합니다. </li><li> 그래서 미래의 모든 이미지 조정이 고르게에 적용 할 이미지를 평평이미지. 에이 클릭하려면 : 프로세스 탭, 2D 필터를 평평하게, &quot;밝은,&quot;개체 폭의 BG 강도는 &quot;75,&quot;적용 할 수 있습니다. </li><li> 모세 혈관이 최대로 가시화되도록 영상의 콘트라스트를 올린다. 이렇게하려면 클릭 : 75에 대비 인상, 탭 표시를 조정합니다. </li><li> 프로세스 탭, 2D 필터, 반점 제거, 커널 크기 7 × 7을 클릭하여 모세 혈관의 가장자리를 부드럽게 이미지를 반점 제거, 적용됩니다. </li><li> 모세관은 검은 색 및 흰색 배경하므로 화상의 명암을 마무리. &quot;가장 적합한&quot;모델 히스토그램을 조정하여이 단계를 수행합니다. 참고 : 처리 된 이미지는 다음 단계를 다음과 같이해야하는지의 예를 들어 대표 결과에 그림 2 참조하시기 바랍니다. </li></ol> 모세관 수를 수행 2. / 정량 모세관 밀도 <ol><li> 각 이미지를 수동 &quot;타겟 오브젝트&quot;를 이용 featur 잘 정의 모세관의 한 부분을 선택객체가 프로그램에 의해 계산하기로 전자 인식합니다. 그리고, 계산 알고리즘에 의해 무시 될 필요 영역에 참조로, &quot;배경&quot;기능을 이용하여, 배경의 작은 부분을 선택한다. 주의 : 이러한 하이라이트 조합 배경 잡음을 무시하면서 모든 모세관이 강조되게. 정량 (카운팅) 프로토콜은 컬러 및 형태에 기초하여 상기 이미지의 부분을 구별하는 컴퓨터 기능을 이용한다. </li><li> 즉시 영상 장비와 이미지의 모든 모세 혈관을 계산하는 카운트 기능을 사용합니다. 모세관으로 배경 잡음을 계산 피하기 위해 5 픽셀 카운트 오브젝트의 최소 직경을 설정한다. </li><li> 보다 신뢰성 평가를 얻기 위해서는 4 이미지 - 각 개인에 대한, (3)의 평균을 계산. 참고 : 이미지가 계산 과정에서 같이해야하는지의 예를 들어 대표 결과에 그림 3을 참조하십시오. </li></ol&gt; 3. 작성 및 이미지 처리를 자동화하는 매크로를 사용하여 주 : 시간을 절약하기 위해, 매크로 자동 하나 또는 많은 이미지에 특정 프로세스 시퀀스를 수행하기 위해 생성 될 수있다. 이 시퀀스는 이미지 수정 빨리 만들기 위해 사용자 정의 할 수 있습니다. 본질적으로,이 매크로는 이미지가 처리되는 방식을 기억하고 신속하고 사용자 입력없이 모든 단계를 수행합니다. 매크로없이 사진 당 약 8 분 반대로 12 모세관 이미지에 카운트를 수행하면, 매크로 (2 사진 당 분 3)과 20 ~ 30 분 사이에이 실습을합니다. 따라서 매크로를 사용하여 3 ~ 5 배 더 효율적 수동으로 각각의 이미지를 처리​​하는 것보다. <ol><li> 매크로를 만들려면, &quot;기록 매크로&quot;를 선택하고 하나의 이미지에 위의 1 단계와 2 단계에 설명 된대로 단계 및 프로세스가 원하는 수행합니다. 나중에 참조 할 수 있도록 수행 된 이미지 처리 단계에 따라 매크로 이름을 지정합니다. 때 USI를미래의 사진에 매크로를 겨, 단순히 &quot;매크로 실행&quot;버튼을 클릭 프로그램이 자동으로 원하는 이미지 (들)에 기록 된 개선 사항을 적용합니다. 참고 :이 연구소는 몇 초에서 방법의 제 1 단계 중 하나를 제외하고 모두 수행하기 위해 매크로를 사용합니다. 사용자 입력을 필요로하는 유일한 단계는 이미지를 자르고, 여기서 단계 1.2을 선택한다. </li></ol>

<ol>
	<li>Gronenschild, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Semi-automatic+assessment+of+skin+capillary+density:+proof+of+principle+and+validation.">Semi-automatic assessment of skin capillary density: proof of principle and validation.</a> Microvasc Res. 90, 192-198 (2013).</li><li>Allen, J., Howell, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvascular+imaging:+techniques+and+opportunities+for+clinical+physiological+measurements.">Microvascular imaging: techniques and opportunities for clinical physiological measurements.</a> Physiol Meas. 35, R91-R141 (2014).</li><li>Boettcher, M., Gloe, T., de Wit, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Semiautomatic+quantification+of+angiogenesis.">Semiautomatic quantification of angiogenesis.</a> J Surg Res. 162, 132-139 (2010).</li><li>Wild, R., Ramakrishnan, S., Sedgewick, J., Griffioen, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+angiogenesis+and+tumor+vessel+architecture+by+computer-assisted+digital+image+analysis:+effects+of+VEGF-toxin+conjugate+on+tumor+microvessel+density.">Quantitative assessment of angiogenesis and tumor vessel architecture by computer-assisted digital image analysis: effects of VEGF-toxin conjugate on tumor microvessel density.</a> Microvasc Res. 59, 368-376 (2000).</li><li>Tresadern, P. 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=Simulating+nailfold+capillaroscopy+sequences+to+evaluate+algorithms+for+blood+flow+estimation.">Simulating nailfold capillaroscopy sequences to evaluate algorithms for blood flow estimation.</a> Conf Proc IEEE Eng Med Biol Soc. , 2636-2639 (2013).</li><li>Anderson, M. 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=Computerized+nailfold+video+capillaroscopy--a+new+tool+for+assessment+of+Raynaud's+phenomenon.">Computerized nailfold video capillaroscopy--a new tool for assessment of Raynaud's phenomenon.</a> J Rheumatol. 32, 841-848 (2005).</li><li>Neubauer-Geryk, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+reactivity+of+skin+microcirculation+in+response+to+L-arginine+in+later-onset+type+1+diabetes.">Decreased reactivity of skin microcirculation in response to L-arginine in later-onset type 1 diabetes.</a> Diabetes Care. 36, 950-956 (2013).</li><li>Pazos-Moura, C. C., Moura, E. G., Bouskela, E., Torres-Filho, I. P., Breitenbach, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nailfold+capillaroscopy+in+diabetes+mellitus:+morphological+abnormalities+and+relationship+with+microangiopathy.">Nailfold capillaroscopy in diabetes mellitus: morphological abnormalities and relationship with microangiopathy.</a> Braz J Med Biol Res. 20, 777-780 (1987).</li><li>Antonios, T. F., Singer, D. R., Markandu, N. D., Mortimer, P. S., MacGregor, 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=Structural+skin+capillary+rarefaction+in+essential+hypertension.">Structural skin capillary rarefaction in essential hypertension.</a> Hypertension. 33, 998-1001 (1999).</li><li>Kaiser, S. E., Sanjuliani, A. F., Estato, V., Gomes, M. B., Tibirica, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antihypertensive+treatment+improves+microvascular+rarefaction+and+reactivity+in+low-risk+hypertensive+individuals.">Antihypertensive treatment improves microvascular rarefaction and reactivity in low-risk hypertensive individuals.</a> Microcirculation. 20, 703-716 (2013).</li><li>Cheng, C., Diamond, J. J., Falkner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+capillary+rarefaction+in+mild+blood+pressure+elevation.">Functional capillary rarefaction in mild blood pressure elevation.</a> Clinical and Translational Science. 1, 75-79 (2008).</li><li>Debbabi, H., Bonnin, P., Levy, B. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+blood+pressure+control+with+perindopril/indapamide+on+the+microcirculation+in+hypertensive+patients.">Effects of blood pressure control with perindopril/indapamide on the microcirculation in hypertensive patients.</a> Am J Hypertens. 23, 1136-1143 (2010).</li><li>He, F. J., Marciniak, M., Markandu, N. D., Antonios, T. F., MacGregor, 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=Effect+of+modest+salt+reduction+on+skin+capillary+rarefaction+in+white,+black,+and+Asian+individuals+with+mild+hypertension.">Effect of modest salt reduction on skin capillary rarefaction in white, black, and Asian individuals with mild hypertension.</a> Hypertension. 56, 253-259 (2010).</li><li>Murray, A. K., 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+influence+of+measurement+location+on+reliability+of+quantitative+nailfold+videocapillaroscopy+in+patients+with+SSc.">The influence of measurement location on reliability of quantitative nailfold videocapillaroscopy in patients with SSc.</a> Rheumatology (Oxford). 51, 1323-1330 (2012).</li><li>Ingegnoli, F., 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=Feasibility+of+different+capillaroscopic+measures+for+identifying+nailfold+microvascular+alterations.">Feasibility of different capillaroscopic measures for identifying nailfold microvascular alterations.</a> Semin Arthritis Rheum. 38, 289-295 (2009).</li><li>Debbabi, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+skin+capillary+density+in+treated+essential+hypertensive+patients.">Increased skin capillary density in treated essential hypertensive patients.</a> Am J Hypertens. 19, 477-483 (2006).</li><li>Serne, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+skin+capillary+recruitment+in+essential+hypertension+is+caused+by+both+functional+and+structural+capillary+rarefaction.">Impaired skin capillary recruitment in essential hypertension is caused by both functional and structural capillary rarefaction.</a> Hypertension. 38, 238-242 (2001).</li><li>Shore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillaroscopy+and+the+measurement+of+capillary+pressure.">Capillaroscopy and the measurement of capillary pressure.</a> Br J Clin Pharmacol. 50, 501-513 (2000).</li><li>Rieder, M. J., O'Drobinak, D. M., Greene, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+computerized+method+for+determination+of+microvascular+density.">A computerized method for determination of microvascular density.</a> Microvasc Res. 49, 180-189 (1995).</li><li>Vermeulen, P. B., 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=Quantification+of+angiogenesis+in+solid+human+tumours:+an+international+consensus+on+the+methodology+and+criteria+of+evaluation.">Quantification of angiogenesis in solid human tumours: an international consensus on the methodology and criteria of evaluation.</a> Eur J Cancer. 32A, 2474-2484 (1996).</li><li>Cheng, C., Daskalakis, C., Falkner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+Assessment+of+Microvascular+and+Endothelial+Function.">Non-invasive Assessment of Microvascular and Endothelial Function.</a> Journal of Visualized Experiments. , (2012).</li><li>Antonios, T. F., 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=Maximization+of+skin+capillaries+during+intravital+video-microscopy+in+essential+hypertension:+comparison+between+venous+congestion,+reactive+hyperaemia+and+core+heat+load+tests.">Maximization of skin capillaries during intravital video-microscopy in essential hypertension: comparison between venous congestion, reactive hyperaemia and core heat load tests.</a> Clin Sci (Lond). 97, 523-528 (1999).</li></ol>

The authors have no conflicts of interest.

Nailfold capillaroscopy 다양한 종양, 심혈관 질환 및 류마티스 질환 응용 프로그램에 대한 미래에 임상 적으로 유용한 도구로 약속을 보여줍니다. 이미지 획득 과정은 연구자 중에서 상당히 일관성이 아직 영상 처리 및 분석을위한 다수의 방법이 현재 존재한다. 방법은 현재 컴퓨터 및 수동 모세관 수를 포함한다. 수동 계산은 시간 소모적이기 때문에 문제가 있고, 사용자의 주관에 따라 피로. 현재 컴?...

미세 혈관 이미징. 많은 잠재적 임상 적용에 급속히 성장하는 분야, 예를 들면 2, 종양 가능한 치료 방법으로 종양과 통찰력의 상태에 관한 유용한 정보를 산출, 종양 혈관 신생의 범위를 결정하는 미세 혈관 이미지를 사용하고있다. (3) (4) 단, nailfold의 capillaroscopy 아마도 미세 혈관 이미징의 가장 비용 효율적이고 광범위하게 적용 할 수있는 형태이다. 연구진은 모세관 형태를 혈류 속도를 연구하고 조사하는 비디오 nailfold의 capillaroscopy을 사용하고 있습니다. (5) (6) 비디오 및 정지 영상 nailfold의 capillaroscopy 모두를하고 있습니다 부속 진단 및 전신 경화증으로 레이노 현상과 다양한 결합 조직 질환 치료를위한 신경. 2 Nailfold capillaroscopy뿐만 아니라 다양한 잠재적 인 심장 혈관 애플리케이션을 보유하고 있습니다. 현재 연구 사용 nailfold의 capillaroscopy 시사그 당뇨병 유형 1과 유형 2의 환자는 당뇨병이 아닌 개인들에 비해 불변 모세관 밀도 이상 모세관 형태의 높은 유병률을 나타내고, 아직. 7-8 Capillaroscopy 또한 고혈압 실험적으로 연구되고있다. 감소 모세관 밀도로 이어지는 구조 모세관 진공이 아닌 고혈압 사람들에 비해 고혈압 개인에서 입증되었다. 9-10 이러한 이전 고혈압 환자는 달리 구조 진공을 전시, 최근의 연구가 있음을 보여 주었다 (40 위의 평균 연령) 젊은 고혈압 환자는 구조 진공없이 기능 진공이 (세 미만 40 세 의미). (11)이 기능 진공 전에 발생 제안 및 구조 진공 시간에 걸쳐 진행된다. 흥미롭게도, 고혈압 환자는 페린도 프릴과 같은 특정 항 고혈압 약물로 치료 / 인다 파 미드는 규범을 표시알 모세관 밀도와 처리 후의 내피 기능, ACE (안지오텐신 전환 효소) 억제제 또는 이뇨제로 치료 것과 필적 혈압 조절에도 불구하고 낮은 모세관 밀도를 유지한다. (12)이 일부 항 고혈압 약물 모세관을 역전시킴으로써 모세관 밀도를 정상화 수 있음을 시사 고혈압에 의한 진공. 또한, 다른 연구자는 소금 섭취 감소 고혈압 개인의 기능적 및 구조적 모세관 희박 반전 리드 것으로 나타났다. 13 이 기술의 다양한 잠재적 인 임상 적용에도 불구하고, 기술에 약간의 표준화가 모세 혈관 밀도 이미지를 정량을 위해 존재한다. (2) 지금까지 연구자들은 모세 혈관 밀도 결과 내 관찰자 간 관찰자의 관점 만 정확한을 경우 모두에서 재현 할 수 있음을 발견했다 같은 지역 각 시간을 계산하고있다. 1,14 (15) </sup&gt; 참고로, 이전의 연구는 크게 수동으로 느리고 주관적인 과정이다 육안, 9 16 17 18를 사용하여 모세관 카운트를 수행했습니다. 모세관 이미지의 정량을위한 표준화, 컴퓨터 기반의 알고리즘은 이론적 capillaroscopy의 임상 적용을 용이하게, 덜 주관으로보다 효율적이고 재현성있는 데이터 분석을 제공합니다. 일부 연구자들은 참으로. nailfold capillaroscopic 사진에서 데이터를 정량화하기 위해 1,6 19 20 그러나, 현재까지 단 하나의 게시 capillaroscopy 데이터, 1 정량에 사용할 수 복잡한 소프트웨어 프로그램의 신뢰성에 대해 설명을 컴퓨터 기반의 프로그램을 사용하고이 프로그램은 다음과 같이 복잡 이전에 동일한 시야를 계산하기위한 요구 사항에 의해 위에서 언급. 여기서는 허용 표준 알고리즘을 이용하여 정량 모세관을위한 간단하고 신뢰성있는 프로토콜을 제시복수의 시야의 용도. 복수의 시야의 사용뿐만 아니라 절차를 단순화 할뿐만 아니라 모세관 카운트 정상적인 생물학적 변화의 평가를 허용한다. 본 연구의 목적은 모세관 정량 처리를 표준화 및 재현성 효율적인 컴퓨터 기반의 알고리즘을 설명하기위한 것이다. 이러한 방법은 완전히 자동화되지 않지만 그들은 매우 작은 사용자 입력을 필요로하고, 이미지의 신속하고 신뢰성있는 정량을 제공한다.

<table>
	<tbody>
		<tr>
			<td class="xl65">Image-Pro Premier</td>
			<td class="xl65">Media Cybernetics, Inc</td>
			<td align="right" class="xl63">9.1</td>
			<td class="xl63">Image processing software</td>
		</tr>
	</tbody>
</table>

a reproducible computerized method for quantitation of capillary density using nailfold capillaroscopy

Capillaroscopy 직접 미세 순환을 시각화, 비 침습적 효율이 상대적으로 저렴하고 쉽게 배울 방법론이다. capillaroscopy 기술은 잠재적으로 가치있는 피부 질환, 안과, 류마티스 및 심혈관 임상 다양한 용도로 이어지는, 환자의 미세 혈관의 건강에 대한 통찰력을 제공 할 수있다. 또한, 종양의 성장은 종양 내 미세 혈관 밀도를 측정함으로써 정량화 될 수있는 혈관 신생에 의존 할 수있다. 그러나,이 기술없이 표준화에 현재 조금이고, 지금까지 단 하나의 공보 capillaroscopy 데이터 정량을 위해 현재 사용 가능한 복잡한 컴퓨터 기반 알고리즘의 신뢰성을보고한다. (1)이 논문은 새로운 간단하고 신뢰할 표준화 모세관 카운팅 알고리즘을 설명 nailfold capillaroscopy 데이터를 정량합니다. 간단하고 재현성 컴퓨터 capillaroscopy 알고리즘 등이 더 용이하게하는 것처럼연구자와 임상의 간의 기술의 광범위한 사용. 많은 연구자들은 현재 결과 사용자의 피로와 주체성을 촉진, 손으로 capillaroscopy 이미지를 분석. 이 논문은 신규 한 설명하기 쉬운 화상 처리 알고리즘 재현성, 반자동 카운팅 알고리즘 이외에 자동화. 이 알고리즘은 주관성을 감소시키면서 분 화상의 분석을 가능하게; 트레이닝 시간의 최소량은 (우리의 경험에서, 1 시간 미만)의 기술을 배울 필요가있다.

이 화상 처리 순서의 목적은 정확하게 정량화 될 수 있도록 배경 이미지에서 모세관을 구별하는 것이다. 모두 불완전한 화상 처리 과도한 화상 처리는 모세관을 정량화하는 프로그램의 기능에 해롭다. 도 3에 도시 된 바와 같이, 불완전한 화상 처리는 배경으로부터 구별 모세관 어렵게한다. 이는 사용자가 상기 계산 방법은 정확하게 몇 모세관을 강조 할 수있는 사용자의 능력에 의존...

Watch this Scientific Journal Video about A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy at JoVE.com

A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm.

Nailfold Capillaroscopy를 사용하여 모세관 밀도의 정량에 대한 재현 전산 방법

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy to learn methodology for directly visualizing the microcirculation. The ...

a-reproducible-computerized-method-for-quantitation-capillary-density

Research

JoVE Journal

Medicine

8.9K 조회수.  Thomas Jefferson University. Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm. 

비디오: A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombotic 국소 빈혈 : 마우스 스트로크 연구를위한 최소 침습 및 재현성 광화학 대뇌 피질의 병변 모델

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

연구

의학

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

생성 및 마우스의 동맥 병변의 3 차원 정량 광학 프로젝션 단층 촬영을 사용하여

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic conditions. As part of a diagnostic assessment, the early detection of postural deficits is important so that appropriate and targeted interventions can be prescribed. The Sensory Organization Test (SOT) on the EquiTest determines an individual&#39;s use of the sensory systems (somatosensory, visual, and vestibular) that are responsible for postural control. Somatosensory and visual input are altered by the calibrated sway-referenced support surface and visual surround, which move in the anterior-posterior direction in response to the individual&#39;s postural sway. This creates a conflicting sensory experience. The Motor Control Test (MCT) challenges postural control by creating unexpected postural disturbances in the form of backwards and forwards translations. The translations are graded in magnitude and the time to recover from the perturbation is computed.
Intermittent claudication, the most common symptom of peripheral arterial disease, is characterized by a cramping pain in the lower limbs and caused by muscle ischemia secondary to reduced blood flow to working muscles during physical exertion. Claudicants often display poor balance, making them susceptible to falls and activity avoidance. The Ankle Brachial Pressure Index (ABPI) is a noninvasive method for indicating the presence of peripheral arterial disease and intermittent claudication, a common symptom in the lower extremities. ABPI is measured as the highest systolic pressure from either the dorsalis pedis or posterior tibial artery divided by the highest brachial artery systolic pressure from either arm. This paper will focus on the use of computerized dynamic posturography in the assessment of balance in claudicants.

Individuals with intermittent claudication exhibit poor balance compared to healthy controls. Computerized dynamic posturography is an objective method for measuring an individual's postural responses to balance disturbances. This provides an objective reflection of the person&#x2019;s ability to respond to situations under which sensory stimuli are altered and unexpected perturbations occur.

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

복강 내 난소 암 전이의 정량

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

쥐 간 문부 클램프 모델을 사용하여 직접 분절 인트라 간 납품 방법

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

쥐 모델에서 Infrarenal 복부 대동맥 클램핑에 의해 재현 동맥 발가 벗김 부상

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Method for Quantifying Upper Limb Performance in Daily Life Using Accelerometers

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. It has become important to measure a person's activities in daily life, and not just measure their capacity for activity in the structured environment of a clinic or laboratory. A wearable sensor that is now enabling measurement of daily movement is the accelerometer. Accelerometers are commercially-available devices resembling large wrist watches that can be worn throughout the day. Data from accelerometers can quantify how the limbs are engaged to perform activities in peoples' homes and communities. This report describes a methodology to collect accelerometry data and turn it into clinically-relevant information. First, data are collected by having the participant wear two accelerometers (one on each wrist) for 24 h or longer. The accelerometry data are then downloaded and processed to produce four different variables that describe key aspects of upper limb activity in daily life: hours of use, use ratio, magnitude ratio, and the bilateral magnitude. Density plots can be constructed that visually represent the data from the 24 h wearing period. The variables and their resultant density plots are highly consistent in neurologically-intact, community-dwelling adults. This striking consistency makes them a useful tool for determining if upper limb daily performance is different from normal. This methodology is appropriate for research studies investigating upper limb dysfunction and interventions designed to improve upper limb performance in daily life in people with stroke and other patient populations. Because of its relative simplicity, it may not be long before it is also incorporated in clinical neurorehabilitation practice.

This protocol describes a method to quantify upper limb performance in daily life using wrist-worn accelerometers.

가속도계를 사용하여 일상 생활에서 상지 성능을 정량화하는 방법

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. ...

Ablation of Ischemic Ventricular Tachycardia Using a Multipolar Catheter and 3-dimensional Mapping System for High-density Electro-anatomical Reconstruction

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars represent zones of slow conduction that allow the occurrence and maintenance of reentrant circuits. Catheter ablation enables substrate modification of those low voltage areas and thus can help to alter the scar tissue in such a way that arrhythmias cannot appear anymore. Hospitalizations of concerned patients decrease, quality of life and outcome rise. Consequently, VT ablation represents a growing field in electrophysiology, especially for patients with endocardial scars in ischemic heart disease after myocardial infarction. However, ablation of ventricular tachycardia remains one of the most challenging procedures in the electrophysiology lab. Precise scar definition and localization of abnormal potentials are critical for ablation success. The following manuscript describes the use of a multipolar mapping catheter and 3-dimensional (3D) mapping system to create a high density electro-anatomical map of the left ventricle including a precise scar representation as well as mapping of fractionated and late potentials in order to allow a highly accurate substrate modification.

With the following protocol, we provide an approach to Ventricular Tachycardia (VT) ablation using high density mapping with a multipolar catheter and 3D mapping system enhancing the success of the procedure.

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars ...

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a computerized functional skills training program that was paired in half of the participants with a commercially available cognitive training (CCT) program.
Non-impaired older individuals (NC) aged 60+ (n=45) and similarly aged individuals with mild cognitive impairment (MCI; n=50) were randomized to receive 12 weeks of twice-weekly computerized functional skills training (CFST) or 12 weeks of twice-weekly sessions split between CCT and CFST. Skills trained were use of an ATM; internet banking; ticket kiosk; telephone and internet prescription refill; medication management; and internet shopping. As with previous functional capacity assessments, we focus on completion time for each simulation.
51 participants completed the training program, either by mastering all 6 tasks (34) or completing 12 weeks of training. 44 more participants completed 4 or more training sessions so they were also analyzed for improvement up to their last training session. Completion time for all 6 tests significantly improved from the baseline assessment to the final training session in both groups of participants (all p&#60;0.001 with an average improvement in task completion time of 45%). Further, there was no differential improvement in MCI and NC in the 6 tests from baseline to end of training (all t&#60;1.66, all p&#62;0.12). Finally, combined CCT plus CFST did not differ from CSFT alone on any of the percent-change score measures (all t&#60;1.64, all p&#62;0.11).
Both NC and MCI groups evidenced substantial improvements in performance. CCT supplementation led to similar functional gains with half as many training sessions. The NC participants proceeded through the training fairly rapidly even without CCT supplementation; MCI participants required more training but learned equivalently. These findings suggest that even in cases with memory impairments, functional skills can be efficiently learned with training.

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.

전산화된 기능 기술 평가 및 교육 프로그램 대상 기술 기반 일상 기능 기술

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a ...

A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ranges between 20 and 30%. Novel treatment approaches in order to reduce sepsis-related multiorgan failure and death are urgently needed. Robust animal models allow for one or multiple treatment approaches as well as for testing their effect on physiological and molecular parameters. In this article, a simple animal model is presented.
First, general anesthesia is induced in animals either with the use of volatile or by intraperitoneal anesthesia. After placement of an intravenous catheter (tail vein), tracheostomy, and insertion of an intraarterial catheter (tail artery), mechanical ventilation is started. Baseline values of mean arterial blood pressure, arterial blood oxygen saturation, and heart rate are recorded.
The injection of lipopolysaccharides (1 milligram/kilogram body weight) dissolved in phosphate-buffered saline induces a strong and reproducible inflammatory response via the toll-like receptor 4. Fluid corrections as well as the application of norepinephrine are performed based on well-established protocols.
The animal model presented in this article is easy to learn and strongly oriented towards clinical sepsis treatment in an intensive care unit with sedation, mechanical ventilation, continuous blood pressure monitoring, and repetitive blood sampling. Also, the model is reliable, allowing for reproducible data with a limited number of animals in accordance with the 3R (reduce, replace, refine) principles of animal research. While animal experiments in sepsis research cannot easily replaced, repetitive measurements allow for a reduction of animals and keeping septic animals anesthetized diminishes suffering.

Here, we present a reproducible intensive care unit-oriented endotoxin model in rats.

쥐의 재현 가능한 집중 치료실 지향 내독소 모델

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ...