이 연구는 국립 과학 재단 (National Science Foundation) 부여 번호 0651981과 알렉산더와 마가렛 스튜어트 신뢰 존스 홉킨스 대학 불구하고 암 센터 투자했다. 저자는 IRB와 환자의 연구뿐만 아니라 도움 박사 세원 강과 환자의 연구 기간 동안 자신의 부서를 지원하는 박사 로다 알라니의 기여를 인정하고 싶습니다.

1. 설정 <ol><li> 적외선 이미지 수집 및 저장뿐만 아니라 컴퓨터에 연결된 데이터 수집 카드 적외선 카메라와 PC를 갖춘 온도 제어 시험 실 Fig.1a에 표시됩니다. </li><li> 상온 및 피부 표면 온도가 환자의 연구 및 측정 데이터 동안 데이터 수집 카드에 연결된 열전쌍으로 모니터링하는 것은 컴퓨터에 저장됩니다. </li></ol> 2. 이미지 수집 <ol><li> 병변이 냉각 효과없이 이미지에서 열 감지되지 않을 수 있기 때문에 사각형 접착제 표시는 관심과 그 주변의 색소 병변을 (그림의 1B) 집중하는 데 사용됩니다. </li><li> 우리는 디지털 카메라 (캐논 파워샷 G11) (그림의 1B)와 색소 병변과 접착 윈도우의 밝은 빛을 이미지를 획득. </li><li> 디지털 카메라 (DermLite Foto 시스템)에 연결 dermatoscope는 편광 이미지를 캡처하는 데 사용됩니다. </li><li> 우리는 멀린 midwave (3-5 μm의) Fig.1a, C. 표시 적외선 카메라와 함께 안정된 상태 적외선 이미지를 획득 </li><li> 우리는 1 분 동안 병변뿐만 아니라 50mm 직경 주변 지역을 포함하는 환자의 피부의 영역에 차가운 공기의 흐름을 적용합니다. </li><li> 일분 후, 우리는 피부를 3-4 분 이내에 실내 온도 (열 복구 단계) (그림 1C - D)에 다시 따뜻한 수 있도록하려면이 냉각 스트레스를 제거합니다. </li><li> 열 복구 단계 동안, 색소 병변의 적외선 이미지는 매 2 초 (그림 1C - D)를 캡처합니다. </li><li> 연구 기간 동안 찍은 모든 IR 이미지 (흰색 빛이 편광 이미지 이외에) 저장되고 Labview 소프트웨어를 사용하여 저장됩니다. </li></ol> 3. 이미지 처리 <ol><li> IR 이미지는 피부 표면에 정확하게 과도 온도 배포판을 얻기 위해서는 전용 Matlab 코드를 사용하여 분석하고 있습니다. 이러한 목적을 위해, 우리는 몇 가지 보정 단계와 multimodal 영상 분석 시스템을 소개합니다. </li><li> 우리는 접착제 마커의 구석을 번역에 대한 밝은 빛을 이미지로 획기적인 탐지 알고리즘을 적용 시작. 다음, 우리는 참조 IR 이미지의 해당 지점을 확인. </li><li> 환자의 불수의 본문 / 사지 움직임 보상하기 위해, 우리는 복구 단계에서 IR 이미지 시퀀스를 정렬을위한 이차 모션 모델의 랜드마크로서이 포인트를 사용합니다. </li><li> 우리는 무작위 워커, 사용자가 공간 병변을 delineating 마스크 이미지를 만들 수, 씨앗 포인트를 배치하여 세그먼트 가이드 수있는 대화형 이미지 분할 알고리즘을 사용합니다. </li><li> 일단 우리가 병변의 모양을 결정, 우리는 등록된 IR 이미지 각 해당 지역을 식별합니다. </li><li> 우리는 각각 병변과 건강한 조직을 대표하는 병변 떨어져 병변 내부의 임의의 지점을 선택합니다. </li><li> 우리는 건강한 피부와 병변의 반응의 과도 열 응답을 비교합니다. </li><li> 디지털, dermoscopy, 색상 코드 냉각 여기 이후 주변 조건과 2 초 기록 병변과 주변 지역의 IR 이미지 및 병변과 건강한 조직의 과도 열 응답 : 우리는 모든 데이터를 보여주는 테이블을 준비합니다. </li></ol> 4. 대표 결과 : <img src="/files/ftp_upload/2679/2679fig1.jpg" alt="그림 1" /> 그림 1. 주위 온도 영역의 색소 병변의 클러스터 및 이미징, C 신청한 템플릿 프레임과 함께 더 큰 신체 표면적의) 임상 시험 실에서 적외선 이미징 시스템 HRIS, B)가 사진)을 참조 적외선 이미지, D) 냉각 및 전자 후에도 같은 지역)이 흑색증의 병변과 주변 부분을 확대 <img src="/files/ftp_upload/2679/2679fig2.jpg" alt="그림 2" /> 그림 2. 우리 열 이미징 시스템과 시험 실. <img src="/files/ftp_upload/2679/2679fig3.jpg" alt="그림 3" /> 그림 3. 소용돌이 관에서 차가운 공기의 흐름을 막아서 병변과 주위 피부 조직을 냉각.

<ol>
	<li>Elder, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+progression,+early+diagnosis+and+prognosis+of+melanoma.">Tumor progression, early diagnosis and prognosis of melanoma.</a> Acta Oncol. 38, 535-547 (1999).</li><li>Wartman, D., Weinstock, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+we+overemphasizing+sun+avoidance+in+protection+from+melanoma.">Are we overemphasizing sun avoidance in protection from melanoma.</a> Cancer Epidemiol Biomarkers Prev. 17, 469-470 (2008).</li><li>Pirtini Cetingul, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Using high resolution infrared imaging to detect melanoma and dysplastic nevi [dissertation]. , (2010).</li><li>Jones, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+reappraisal+of+the+use+of+infrared+thermal+image+analysis+in+medicine.">A reappraisal of the use of infrared thermal image analysis in medicine.</a> IEEE Trans. Med. Imaging. 17, 1019-1027 (1998).</li><li>Anbar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+thermal+imaging+today-shifting+from+phenomenological+thermography+to+pathophysiologically+based+thermal+imaging.">Clinical thermal imaging today-shifting from phenomenological thermography to pathophysiologically based thermal imaging.</a> IEEE Eng. Med. Biol. Mag. 17, 25-33 (1998).</li><li>Anbar, M., Gratt, B. M., Hong, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermology+and+facial+telethermography.+Part+I:+history+and+technical+review.">Thermology and facial telethermography. Part I: history and technical review.</a> Dentomaxillofacial Radiology. 27, 61-67 (1998).</li><li>Jones, B. F., Plassmann, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digital+infrared+thermal+imaging+of+human+skin.">Digital infrared thermal imaging of human skin.</a> IEEE Eng. Med. Bio. 21, 41-48 (2002).</li><li>Qi, H., Diakides, N. 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> Infrared imaging in Medicine. , (2007).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+skin+lesions+from+the+transient+thermal+response+using+infrared+imaging+technique.">Identification of skin lesions from the transient thermal response using infrared imaging technique.</a> IEEE 5th Int. Symp. on Biomedical Imaging: From Nano to Macro 1-4. , 1219-1222 (2008).</li><li>Cetingul, P. i. r. t. i. n. i., M, ., Herman, C. <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+the+thermal+signature+of+a+melanoma+lesion.">Quantification of the thermal signature of a melanoma lesion.</a> Int. Journal of Thermal Science. 50, 421-431 (2011).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+heat+transfer+model+of+skin+tissue+for+the+detection+of+lesions:+sensitivity+analysis.">A heat transfer model of skin tissue for the detection of lesions: sensitivity analysis.</a> Physics in Medicine and Biology. 55, 5933-5951 (2010).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+evaluation+of+skin+lesions+using+transient+thermal+imaging.">Quantitative evaluation of skin lesions using transient thermal imaging.</a> Proc. Int. Heat Transfer Conf. , (2010).</li></ol>

관심 없음 충돌 선언하지 않습니다.

결과는 냉각 스트레스를 적용하여 우리는 병변 주변 건강한 조직 사이의 온도 차이를 개선하는 것이 좋습니다. 또한, 때문에 열 이미징하는 동안 환자의 작은 움직임을 우리는 제대로 참조 상태 및 열 복구하는 동안 온도 분포 사이의 온도 차이를 측정하는 이미지를 오버레이 추적 모션을 적용했다. 모션 추적없이 우리는 악성 병변과 건강한 조직 사이의 온도 차이를 감지하고 측정할 수 없었을 거에요. 정상 상태 정보 혼자 명확하게 동적 열 영상의 장점을 증명을 기반으로 IR 이미징을 사용하여 흑색증를 진단하려고 할 때 이러한 결과와 어려움의 수사 설명 추적 정확한 운동의 필요성은 과거에 직면했다. 이것은 IR 카메라 (IR 초점 평면 배열에있는 픽셀의 수)의 공간적 해상도 안목 작은 병변 때 중요합니다 있다고 지적한다. 공간 해상도와 초기 적외선 카메라의 온도 감도 모두는 과거에 초기 단계의 흑색증를 감지에 어려움을 계정에있는, 제한했다. 우리의 접근 방법과 사전에 열 이미징 시도 사이의 주요 차이점 - 그건 적당히 성공은 - 교정의 시퀀스 우리가 정확하게 활성 냉각에 의존 동적 이미징 프로세스뿐만 아니라이 시스템의 온도 차이를 측정하도록 영상 처리 단계입니다.

<table border="1"><tbody><tr><td> 장비의 명칭 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> 멀린 MWIR 카메라 </td><td> FLIR </td><td> - </td></tr><tr><td> 캐논 파워샷 G11 </td><td> 규범 </td><td> - </td></tr><tr><td> DermLite Foto 시스템 </td><td> DermLite </td><td> - </td></tr><tr><td> 소용돌이 관 </td><td> Exair </td><td> - </td></tr><tr><td> 에어 탱크 </td><td> Airgas </td><td> - </td></tr></tbody></table>

quantitative visualization and detection of skin cancer using dynamic thermal imaging

2010 년 약 68,720 melanomas는 8650 주위에 죽음이 하나의 결과로, 미국 혼자 진단한다. 날짜하려면, 흑색증 수있는 유일한 효과적인 치료 그러므로, 확장 생존의 열쇠가 2,3 조기 발견이며, 수술 절단 남아있다. 환자의 큰 숫자를 고려하면 신속하게 전문적인 치료에 접근 매년과 한계를 진단, 진단을 돕기 위해 생체내 진단 악기의 목적의 개발이 필수적입니다. 피부암, 특히 비침습 진단 도구를 감지하는 새로운 기술은 수많은 실험실에서 살펴되고있다. 디지털 사진, dermoscopy, multispectral 이미징 시스템 (MelaFind), 레이저 기반의 시스템 (공촛점 스캐닝 레이저 현미경, 레이저 도플러 재관류 이미징, 광학 일관성의 tomography), 초음파, 자기 공명 영상으로 수술 방법, 기법, 함께하는 것은 테스트하고 있습니다 . 각 기술은 효율성과 정확성 대 사용의 용이성 및 비용 고려 사항 사이의 타협을 제기하는 많은 독특한 장점과 단점을 제공합니다. 이러한 기술과 비교에 대한 자세한 내용은 문학 4에서 사용할 수 있습니다. 적외선 (IR​​) 이미징은 현지 피부 온도를 측정하여 특정 질병의 징후를 진단하는 유용한 방법으로 표시되었다. 질병이나 일반적인 기능의 편차가 신체의 온도 변화에 의해 수반되는 보여주는 증거의 많은 시체가있다, 이것은 다시 피부 5,6의 온도에 영향을 미칩니다. 인체와 피부의 온도에 대한 정확한 데이터는 종종 질병으로 인한 정상적인 조건에서 특정 편차의 열 생성 및 thermoregulation, 책임이있는 프로세스에 대한 풍부한 정보를 제공할 수 있습니다. 그러나, IR 영상이 널리 7,8 십년 전에 기술의 조기 사용으로 인해 의학에서 인정되지 않은, 온도 측정 정확도 및 공간적 해상도가있을 때 부적 절한하고 정교한 이미지 처리 도구를 사용할 수 있었다. 이러한 상황은 1990 년대 후반 - 2​​000 년대에 극적으로 바뀌었습니다. IR 기기, 디지털 영상 처리 알고리즘과 과학자의 공간뿐만 아니라 분석할 수 동적 IR 이미징의 구현뿐만 아니라, 피부 9 시간적 열 행동 진보, 분야에서 획기적인 허용. 우리의 연구에서, 우리는 이론 및 실험 연구와 함께 IR 영상의 가능성을, 탐험 비용 종양 검출에 대한 생체내 광학 측정 기술, 심사 강조하고 흑색증 10-13의 조기 발견으로, 비침습 효과 . 본 연구에서는, 우리는 생검을위한 임상 표시가있는 색소 병변을 가진 환자가 화상을 위해 선택되는 환자 연구에서 얻은 데이터를 표시합니다. 우리는 건강하고 악성 조직 및 생검 결과와 비교의 데이터 간의 온도 응답의 차이를 비교했다. 우리는 흑색증의 병변의 증가 신진 대사 활동이 동적 적외선 이미징에 의해 감지 수 있습니다 결론을 내렸다.

Watch this Scientific Journal Video about Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging at JoVE.com

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

우리는 증가 신진 대사 활동이 가열 및 냉각 여기에 피부의 과도 열 응답의 측정 같지는 양을 생성과 그 악성 색소 병변을 증명하는 것은 초기에 흑색증 및 기타 피부 암 (대 비 proliferative nevi)의 양적 확인하실 수 있습니다 질병의 단계.

양적 시각화 및 동적 써멀 이미징을 사용하여 피부암의 감지

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

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16.6K 조회수.  The Johns Hopkins University. 우리는 증가 신진 대사 활동이 가열 및 냉각 여기에 피부의 과도 열 응답의 측정 같지는 양을 생성과 그 악성 색소 병변을 증명하는 것은 초기에 흑색증 및 기타 피부 암 (대 비 proliferative nevi)의 양적 확인하실 수 있습니다 질병의 단계.

비디오: Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&amp;#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

전체 혈액 자극 분석을 사용하여 프로 염증성 시토킨 IL - 1β의 정확하고 간단한 측정

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

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Quantitative Analysis of Cancer Metastasis using an Avian Embryo Model

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex process that can involve many tissue types, span variable time periods, and often occur deep within organs, making it difficult to investigate and quantify. In addition, the efficacy of the metastatic process is influenced by multiple steps in the metastatic cascade making it difficult to evaluate the contribution of a single aspect of tumor cell behavior. As a consequence, metastasis assays are frequently performed in experimental animals to provide a necessarily realistic context in which to study metastasis. Unfortunately, these models are further complicated by their complex physiology. The chick embryo is a unique in vivo model that overcomes many limitations to studying metastasis, due to the accessibility of the chorioallantoic membrane (CAM), a well-vascularized extra-embryonic tissue located underneath the eggshell that is receptive to the xenografting of tumor cells (figure 1). Moreover, since the chick embryo is naturally immunodeficient, the CAM readily supports the engraftment of both normal and tumor tissues. Most importantly, the avian CAM successfully supports most cancer cell characteristics including growth, invasion, angiogenesis, and remodeling of the microenvironment. This makes the model exceptionally useful for the investigation of the pathways that lead to cancer metastasis and to predict the response of metastatic cancer to new potential therapeutics. The detection of disseminated cells by species-specific Alu PCR makes it possible to quantitatively assess metastasis in organs that are colonized by as few as 25 cells. Using the Human Epidermoid Carcinoma cell line (HEp3) we use this model to analyze spontaneous metastasis of cancer cells to distant organs, including the chick liver and lung. Furthermore, using the Alu-PCR protocol we demonstrate the sensitivity and reproducibility of the assay as a tool to analyze and quantitate intravasation, arrest, extravasation, and colonization as individual elements of metastasis.

Using quantitative PCR, we demonstrate how the well-established chick CAM model can be used to quantitatively analyze the metastasis of human tumor cells to distant organs.

조류 엠브료 모델을 사용하여 암 전이의 정량 분석

During metastasis cancer cells disseminate from the primary tumor, invade into surrounding tissues, and spread to distant organs. Metastasis is a complex ...

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

In molecular imaging, positron emission tomography (PET) and optical imaging (OI) are two of the most important and thus most widely used modalities1-3. PET is characterized by its excellent sensitivity and quantification ability while OI is notable for non-radiation, relative low cost, short scanning time, high throughput, and wide availability to basic researchers. However, both modalities have their shortcomings as well. PET suffers from poor spatial resolution and high cost, while OI is mostly limited to preclinical applications because of its limited tissue penetration along with prominent scattering optical signals through the thickness of living tissues.
 Recently a bridge between PET and OI has emerged with the discovery of Cerenkov Luminescence Imaging (CLI)4-6. CLI is a new imaging modality that harnesses Cerenkov Radiation (CR) to image radionuclides with OI instruments. Russian Nobel laureate Alekseyevich Cerenkov and his colleagues originally discovered CR in 1934. It is a form of electromagnetic radiation emitted when a charged particle travels at a superluminal speed in a dielectric medium7,8. The charged particle, whether positron or electron, perturbs the electromagnetic field of the medium by displacing the electrons in its atoms. After passing of the disruption photons are emitted as the displaced electrons return to the ground state. For instance, one 18F decay was estimated to produce an average of 3 photons in water5. 
 Since its emergence, CLI has been investigated for its use in a variety of preclinical applications including in vivo tumor imaging, reporter gene imaging, radiotracer development, multimodality imaging, among others4,5,9,10,11. The most important reason why CLI has enjoyed much success so far is that this new technology takes advantage of the low cost and wide availability of OI to image radionuclides, which used to be imaged only by more expensive and less available nuclear imaging modalities such as PET.
 Here, we present the method of using CLI to monitor cancer drug therapy. Our group has recently investigated this new application and validated its feasibility by a proof-of-concept study12. We demonstrated that CLI and PET exhibited excellent correlations across different tumor xenografts and imaging probes. This is consistent with the overarching principle of CR that CLI essentially visualizes the same radionuclides as PET. We selected Bevacizumab (Avastin; Genentech/Roche) as our therapeutic agent because it is a well-known angiogenesis inhibitor13,14. Maturation of this technology in the near future can be envisioned to have a significant impact on preclinical drug development, screening, as well as therapy monitoring of patients receiving treatments.

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Use of Cerenkov Luminescence Imaging (CLI) for monitoring preclinical cancer treatment is described here. This method takes advantage of Cerenkov Radiation (CR) and optical imaging (OI) to visualize radiolabeled probes and thus provides an alternative to PET in preclinical therapeutic monitoring and drug screening.

암 치료 모니터링을위한 Cerenkov 발광 이미징 (CLI)

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

Quantitative Multispectral Analysis Following Fluorescent Tissue Transplant for Visualization of Cell Origins, Types, and Interactions

With the desire to understand the contributions of multiple cellular elements to the development of a complex tissue; such as the numerous cell types that participate in regenerating tissue, tumor formation, or vasculogenesis, we devised a multi-colored cellular transplant model of tumor development in which cell populations originate from different fluorescently colored reporter gene mice and are transplanted, engrafted or injected in and around a developing tumor. These colored cells are then recruited and incorporated into the tumor stroma. In order to quantitatively assess bone marrow derived tumor stromal cells, we transplanted GFP expressing transgenic whole bone marrow into lethally irradiated RFP expressing mice as approved by IACUC. 0ovarian tumors that were orthotopically injected into the transplanted mice were excised 6-8 weeks post engraftment and analyzed for bone marrow marker of origin (GFP) as well as antibody markers to detect tumor associated stroma using multispectral imaging techniques. We then adapted a methodology we call MIMicc- Multispectral Interrogation of Multiplexed cellular compositions, using multispectral unmixing of fluoroprobes to quantitatively assess which labeled cell came from which starting populations (based on original reporter gene labels), and as our ability to unmix 4, 5, 6 or more spectra per slide increases, we&#39;ve added additional immunohistochemistry associated with cell lineages or differentiation to increase precision. Utilizing software to detect co-localized multiplexed-fluorescent signals, tumor stromal populations can be traced, enumerated and characterized based on marker staining.1

Complex tissue masses, from organs to tumors, are composed of various cellular elements. We elucidated the contribution of cellular phenotypes within a tissue utilizing multi-labeled fluorescent transgenic mice in combination with multiparameter immunofluorescent staining followed by spectral unmixing to decipher cell origin as well as cell characteristics based on protein expression.

세포의 기원, 종류, 그리고 상호 작용의 시각화를위한 형광 조직 이식에 따라 정량 멀티 스펙트럼 분석

With the desire to understand the contributions of multiple cellular elements to the development of a complex tissue; such as the numerous cell types that ...

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their translational repression. Changes in miRNA expression have been shown to be involved in the development of all major complex diseases. Furthermore, recent findings showed that miRNAs can be secreted to the extracellular environment and enter the bloodstream and other body fluids where they can circulate with high stability. The function of such circulating miRNAs remains largely elusive, but systematic high throughput approaches, such as miRNA profiling arrays, have lead to the identification of miRNA signatures in several pathological conditions, including neurodegenerative disorders and several types of cancers. In this context, the identification of miRNA expression profile in the cerebrospinal fluid, as reported in our recent study, makes miRNAs attractive candidates for biomarker analysis.
There are several tools available for profiling microRNAs, such as microarrays, quantitative real-time PCR (qPCR), and deep sequencing. Here, we describe a sensitive method to profile microRNAs in cerebrospinal fluids by quantitative real-time PCR. We used the Exiqon microRNA ready-to-use PCR human panels I and II V2.R, which allows detection of 742 unique human microRNAs. We performed the arrays in triplicate runs and we processed and analyzed data using the GenEx Professional 5 software.
Using this protocol, we have successfully profiled microRNAs in various types of cell lines and primary cells, CSF, plasma, and formalin-fixed paraffin-embedded tissues.

We describe a protocol of real time PCR to profile microRNAs in the cerebrospinal fluid (CSF). With the exception of RNA extraction protocols, the procedure can be extended to RNA extracted from other body fluids, cultured cells, or tissue specimens.

정량 실시간 PCR을 사용하여 뇌척수액의 마이크로 RNA 프로파일

MicroRNAs (miRNAs) constitute a potent layer of gene regulation by guiding RISC to target sites located on mRNAs and, consequently, by modulating their ...

Measurement of Dynamic Scapular Kinematics Using an Acromion Marker Cluster to Minimize Skin Movement Artifact

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to clearly describe the acromion marker cluster (AMC) method of determining scapular kinematics when using a passive marker motion capture system, with consideration for the sources of error which could affect the validity and reliability of measurements. The AMC method involves placing a cluster of markers over the posterior acromion, and through calibration of anatomical landmarks with respect to the marker cluster it is possible to obtain valid measurements of scapular kinematics. The reliability of the method was examined between two days in a group of 15 healthy individuals (aged 19-38 years, eight males) as they performed arm elevation, to 120&#176;, and lowering in the frontal, scapular and sagittal planes. Results showed that between-day reliability was good for upward scapular rotation (Coefficient of Multiple Correlation; CMC = 0.92) and posterior tilt (CMC = 0.70) but fair for internal rotation (CMC = 0.53) during the arm elevation phase. The waveform error was lower for upward rotation (2.7&#176; to 4.4&#176;) and posterior tilt (1.3&#176; to 2.8&#176;), compared to internal rotation (5.4&#176; to 7.3&#176;). The reliability during the lowering phase was comparable to results observed during the elevation phase. If the protocol outlined in this study is adhered to, the AMC provides a reliable measurement of upward rotation and posterior tilt during the elevation and lowering phases of arm movement.

This report presents details of how to adopt the acromion marker cluster method of obtaining scapular kinematics when using a passive marker motion capture device. As has been described in the literature, this method provides a robust, non-invasive, three-dimensional, dynamic and valid measurement of scapular kinematics, minimizing skin movement artifact.

피부 운동 아티팩트를 최소화하기 위해 견봉 마커 클러스터를 사용하여 동적 스카풀라 운동학의 측정

The measurement of dynamic scapular kinematics is complex due to the sliding nature of the scapula beneath the skin surface. The aim of the study was to ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.

동소 이식 췌장 암 마우스 모델의 동적 대비 강화 된 자기 공명 영상

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe ...

Diffuse Optical Spectroscopy for the Quantitative Assessment of Acute Ionizing Radiation Induced Skin Toxicity Using a Mouse Model

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and negatively impact patient quality of life and long term survival. Advances in the understanding of the biological pathways associated with normal tissue toxicities have allowed for the development of interventional drugs, however, current response studies are limited by a lack of quantitative metrics for assessing the severity of skin reactions. Here we present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe the instrumentation design of the DOS system as well as the inversion algorithm for extracting the optical parameters. Finally, to demonstrate clinical utility, we present representative data from a pre-clinical mouse model of radiation induced erythema and compare the results with a commonly employed visual scoring. The described DOS method offers an objective, high through-put evaluation of skin toxicity via functional response that is translatable to the clinical setting.

We present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe DOS instrumentation design, optical parameters extraction algorithms and the animal handling procedures required to yield representative data from a pre-clinical mouse model of radiation induced erythema.

마우스 모델을 사용하여 급성 이온화 방사선 유발 피부 독성의 정량적 평가를위한 광학 분광학 확산

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Light Sheet Microscopy에 의한 심근 성 심근염의 Murine 모델에서의 백혈구 침윤의 정량적 시각화

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing continuous remodeling with the competing activity of osteoblasts, which produce the new bone matrix, and osteoclasts, whose function is to eliminate mineralized bone. [18F]-NaF is a positron emission tomography (PET) radiotracer that enables visualization of bone metabolism. [18F]-NaF is chemically absorbed into hydroxyapatite in the bone matrix by osteoblasts and can thus noninvasively detect osteoblastic activity, which is occult to conventional imaging techniques. Kinetic modeling of dynamic [18F]-NaF-PET data provides detailed quantitative measures of bone metabolism. Conventional semi-quantitative PET data, which utilizes standardized uptake values (SUVs) as a measure of radiotracer activity, is referred to as a static technique due to its snapshot of tracer uptake in time.&#160; Kinetic modeling, however, utilizes dynamic image data where tracer levels are continuously acquired providing tracer uptake temporal resolution. From the kinetic modeling of dynamic data, quantitative values like blood flow and metabolic rate (i.e., potentially informative metrics of tracer dynamics) can be extracted, all with respect to the measured activity in the image data. When combined with dual modality PET-MRI, region-specific kinetic data can be correlated with anatomically registered high-resolution structural and pathologic information afforded by MRI. The goal of this methodological manuscript is to outline detailed techniques for performing and analyzing dynamic [18F]-NaF-PET-MRI data. The lumbar facet joint is a common site of degenerative arthritis disease and a common cause for axial low back pain.&#160; Recent studies suggest [18F]-NaF-PET may serve as a useful biomarker of painful facetogenic disease.&#160; The human lumbar facet joint will, therefore, be used as a prototypical region of interest for dynamic [18F]-NaF-PET-MRI analysis in this manuscript.

Quantitative [18F]-Naf-PET-MRI Analysis for the Evaluation of Dynamic Bone Turnover in a Patient with Facetogenic Low Back Pain

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. We present detailed methodologies for performing and analyzing dynamic [18F]-NaF-PET-MRI data in a patient with facetogenic low back pain using the lumbar facet joints as a prototypical region of interest.

정량 [18F]-Naf-PET-MRI 분석 은 얼굴 투박성 요통 환자에서 동적 뼈 회전율의 평가를 위한

Imaging techniques that reflect dynamic bone turnover may aid in characterizing a wide range of bone pathologies. Bone is a dynamic tissue undergoing ...