这项工作是由美国国家癌症研究所的卫生补助R01 CA140257-01的国家研究所的支持。

<html> 注:签署了知情同意的患者。此外，这项研究已在符合人类福祉的所有机构，国家和国际准则进行的。 1，准备电脑<ol><li>打开笔记本电脑，并连接USB从DVI2USB采集卡。 </li></ol> 2，准备监视器<ol><li>连接DVI缆线，以监测和画中画，以允许内窥镜医生从这些画面观看视频，而不是计算机。 </li><li>确保站在显示器上，并设置为DVI。 </li><li>连接画中画奥林巴斯系统背面，然后按奥林巴斯系统上的输入按钮来查看图像的大型显示器安装在墙壁上。 </li></ol> 3，激光二极管驱动器设置<ol><li>检查以确保激光二极管驱动器为OFF。它应该只被打开了几分钟进行成像之前。 </li></ol>e_title"&gt; 4。电源板<ol><li>插上笔记本电脑，处理器，激光二极管驱动器，DVI分配器，并epiphan采集卡插入电源板。 </li></ol> （5）运行MDE广角上的笔记本电脑桌面<ol><li>命中"当前文件夹"。 </li><li>输入病人的数量和缩写，然后按"创建病人文件夹"。 </li></ol> 6，准备盖和过滤器<ol><li>当处理过滤器，请始终使用手套和的Kimwipes，尽量减少油污和杂物的过滤器转移。纱布或酒精棉签可能留下纤维，可以与成像干涉。 </li><li>放置几天的Kimwipes到一个表创建一个平台，放下过滤器和瓶盖。这种消毒面积应在手术过程中容易接触到的肠胃病。 </li><li>放下过滤器和瓶盖，并保持关闭的Kimwipes由使用的过程中。 </li></ol> 7，病人的准备<ol><li>对使用病人的同意的VFI和原黄素染料抵达到内套件之前。 </li><li>位置为病人上消化道内镜检查程序。 </li><li>继续利用多光谱数字显微镜（MDM）7标准的白色光成像。 </li></ol> 8，插入和喷雾原黄素<ol><li>用白色光成像完成后，插入和喷雾原黄素染料在感兴趣的组织。 1-5毫米，应足以取决于Barrett的组织的面积。通过在这一步喷涂原黄素染料，有足够的时间（至少1分钟），给出了VFI前有足够的组织吸收。原黄素，这是根据从美国食品药品管理局（IND 102,217）的研究性新药申请涵盖的是，申请后不久，集中细胞核内的荧光造影剂。虽然VFI无法解析个体细胞形态学，当激光二极管平底锅以上的组织时，反射光使内镜欣赏整体腺morpholoGY。 </li></ol> 9，打开激光二极管<ol><li>接通激光二极管。这样做至少有2分钟的成像开始之前。 </li></ol> 10，准备内窥镜的VFI <ol><li>完全退出内窥镜。 </li><li>前处理内窥镜，确保助手有两对手套上。 </li><li>已内镜保持内窥镜的辅助下一个是谁给的Kimwipe平台上面详细的面前。 </li><li>用的Kimwipes，有助理清洗内窥镜的尖端。一定要轻轻擦拭表面，并沿着清洁内窥镜的侧几厘米更容易处理。 </li><li>清洁后，有助理处置外一双手套的。 </li><li>有助理提上了过滤器。插入过滤器上的短圆筒状突出部插入在内窥镜的其互补孔。保持内窥镜垂直，以帮助处理。 </li><li>抱着FILT呃到位，滑动盖在过滤器并将它推下来在内窥镜的尖端。确保盖子是安全的，完全推在与内窥镜冲洗的边缘。确保盖的唇稍微延伸超过内窥镜的前端。最后，确保过滤器仍然存在，并与内窥镜的尖端冲洗。 </li></ol> 11，将内窥镜回食道和图像 12，从食道取出内窥镜<ol><li>使用的Kimwipes小心地将盖和滤除内窥镜。扔掉帽，并清洁过滤器的下一个案例。 </li></ol> 13，清洗过滤器<ol><li>轻轻地清洁过滤器的Kimwipes。 </li><li>填补小杯的CyDex和淹没过滤器12分钟。每隔几分钟把过滤过。 </li><li>轻轻地清洁过滤器的Kimwipes。 </li><li>淹没滤波器，此时在无​​菌水中。 5分钟后，用水枪bOTTLE和轻轻一喷无菌水在过滤器。 </li><li>在过滤器上几的Kimwipes，让它干燥。 </li><li>一旦干燥，地点在之间的Kimwipes储存容器。 </li></ol>

<ol>
	<li>Modiano, N., Gerson, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Barrett's+Esophagus:+Incidence,+etiology,+pathophysiology,+prevention+and+treatment.">Barrett's Esophagus: Incidence, etiology, pathophysiology, prevention and treatment.</a> Therapeutics and Clinical Risk Management. 3, 1035-1145 (2007).</li><li>Brown, L. M., Devesa, S. S., Chow, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Incidence+of+Adenocarcinoma+of+the+esophagus+among+white+Americans+by+sex,+stage,+and+age.">Incidence of Adenocarcinoma of the esophagus among white Americans by sex, stage, and age.</a> Journal of the National Cancer Institute. 100, 1184-1187 (2008).</li><li>Siegel, R., Naishadham, D., Ahmedin, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+Statistics,+2012.">Cancer Statistics, 2012.</a> A Cancer Journal for Clinicians. 62, 10-29 (2012).</li><li>Vieth, M., Ell, C., Gossner, L., May, A., Stolte, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+analysis+of+endoscopic+resection+specimens+from+326+patients+with+Barrett's+esophagus+and+early+neoplasia.">Histological analysis of endoscopic resection specimens from 326 patients with Barrett's esophagus and early neoplasia.</a> Endoscopy. 36, 776-781 (2004).</li><li>Pohl, H., Rosch, T., Vieth, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Miniprobe+confocal+laser+microscopy+for+the+detection+of+invisible+neoplasia+in+patients+with+Barrett's+oesophagus.">Miniprobe confocal laser microscopy for the detection of invisible neoplasia in patients with Barrett's oesophagus.</a> Gut. 57, 1648-1653 (2008).</li><li>Thekkek, N., 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=Modular+video+endoscopy+for+in+vivo+cross-polarized+and+vital-dye+fluorescence+imaging+of+Barrett's-associated+neoplasia.">Modular video endoscopy for in vivo cross-polarized and vital-dye fluorescence imaging of Barrett's-associated neoplasia.</a> Journal of Biomedical Optics. 18, 026007 (2013).</li><li>Roblyer, D., 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=Multispectral+optical+imaging+device+for+in+vivo+detection+of+oral+neoplasia.">Multispectral optical imaging device for in vivo detection of oral neoplasia.</a> Journal of Biomedical Optics. 13, 024019 (2008).</li><li>Egger, 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=Biopsy+surveillance+is+still+necessary+in+patients+with+Barrett's+oesophagus+despite+new+endoscopic+imaging+techniques.">Biopsy surveillance is still necessary in patients with Barrett's oesophagus despite new endoscopic imaging techniques.</a> Gut. 52, 18-23 (2003).</li><li>Thekkek, N., 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=Vital-dye+enhanced+fluorescence+imaging+of+GI+mucosa:+metaplasia,+neoplasia,+inflammation.">Vital-dye enhanced fluorescence imaging of GI mucosa: metaplasia, neoplasia, inflammation.</a> Gastrointestinal Endoscopy. 75, 877-887 (2012).</li><li>Muldoon, T. J., Anandasabapathy, S., Maru, D., Richards-Kortum, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+imaging+in+Barrett's+esophagus:+a+novel,+low-cost+endoscopic+microscope.">High-resolution imaging in Barrett's esophagus: a novel, low-cost endoscopic microscope.</a> Gastrointestinal Endoscopy. 68, 737-744 (2008).</li></ol>

没有什么可以透露。

与标准的内镜检查，在BE肿瘤常常被漏诊8，因为良性上皮化生可以从高度异型增生和腺癌难以区别。作为一种工具，将更好地使胃肠病来解决这个问题目前无法避免的错误，重要的染料增强荧光成像突出了组织的腺体形态从而提供了一个独特的功能来区分组织类型。此外，通过提供的视图长达2.5厘米的领域，VFI使内镜医师能够有效地和有系统地平移，在整个食管远端，使可疑病变更为突出。 为了利用这种新技术，先喷局部造影剂，原黄素，在感兴趣的组织。然后，需要一个495-nm的长通滤波器去除内窥镜后到被牢固地封到MDM小费。最后，重新插入邻近所关注的组织的内窥镜，然后从切换后白色光的激光二极管，绿色荧光提供了在组织的腺框架的体内图像。 在连续荧光欠佳的情况下，采用新鲜的原黄素可以改善的结果，以及开启激光二极管前几分钟的实际使用情况，并调整其电压。另外，对患者安全，使某些帽已被完全推过过滤器，使得过滤器是与内窥镜的顶端齐平。这，反过来，将增强的图像质量通过防止过滤器本身后面粘液积聚。 而VFI能够表征腺形态，一种有形改善白光内窥镜，它不能解决细胞发育不良。也就是说，50〜100微米的空间分辨率使得VFI不能从低度异型增生鉴别良性上皮化生。为了区分可视化细胞TRA这些诊断，更高分辨率的设备它，如核极性和细胞拥挤，需要9,10。作为一个例子，高分辨率显微内窥镜是一个更新颖的技术，可以区分肿瘤组织在细胞水平上为核被照亮明亮的白色和核到细胞质的比例是可以理解的。窄带成像是另一种选择，但它需要使用两个不同的范围和处理器，要么原黄素或荧光的显微内窥镜的后续管理;相比之下VFI允许立即聚焦成像有一个范围和一个处理器，约6-8分钟延长麻醉时间。然而，由于这些器件后者集中于极小的组织区域，从而错过瘤，VFI，除了诊断瘤形成，也可以作为一个宽视场，红色萎靡不振装置用于放置更高分辨率的探测器。因此，内镜筛查Barrett的发育不良的未来将可能涉及的广角侦察相结合的方式枪技术，如VFI，随着细胞分类技术，如显微内窥镜。

在过去的四十年，食管腺癌（EAC）的发病率显著增加1,2;但由于后期诊断，五年生存率小于20％3。在BE内镜检查的现行标准，前体选管会，是白光内镜与段的随机四quandrant镊子&#39;活组织切片检查。不幸的是，这种技术常常射门瘤形成，它可以是平的，细微且难以区分的标准白光成像4。虽然已经在使用激光共聚焦显微镜突出体内细胞功能的成功，病变仍可由于视图的5场减少错过。有一个"桥梁"技术，它可以突出区域进行进一步的共聚焦显微成像将是显着的价值。 因此，增强红旗成像方式，提高到精华素的能力T和活检早期肿瘤在BE将有助于检测选管会在早期，可医治的阶段，并可能导致更有效的治疗，并随后提高生存率。 VFI是一种新型的技术，它结合了高解析度成像上皮外源外用荧光造影，原黄素，突出腺体形态并划定肿瘤（高度异型增生和癌症）在食管远端在改善体内诊断6的希望。经原黄素，该申请后不久，浓缩细胞核内的激发下，荧光图像提供50至100微米的空间分辨率和视高达2.5厘米的字段，允许内镜形象化腺形态。作为一个结果，这个方法使肠胃病学区分经典巴雷特化生，其中有连续的，均匀间隔的腺体和整体均匀的形态，从可以与瘤形成，其具有obliteration的腺体结构。在这里，我们描述了这种新技术的协议与多光谱内窥镜，并提供有代表性的结果证明该设备的描述中良性上皮化生的形态转变为高度异型增生和癌症的效用。

<table><tbody><tr><td>Filter*</td><td>Schott North America, Inc., Duryea, Pennsylvania</td><td>Not Applicable</td><td>495-nm long-pass filter</td></tr><tr><td>Halo Cap &amp;ndash; Medium*</td><td>Barrx Medical</td><td>CP-002A</td><td></td></tr><tr><td>Processor*</td><td>Pentax</td><td>EPK-i</td><td></td></tr><tr><td>Multispectral Digital Microscope**</td><td>Not Applicable</td><td>Not Applicable</td><td></td></tr><tr><td>Kimwipes</td><td>Kimberly-Clark</td><td>KimTech Science</td><td>S-8115</td></tr><tr><td>Cidex</td><td>Advanced Sterilization Products&amp;nbsp;</td><td>CIDEX OPA Solution</td><td></td></tr><tr><td>Proflavine hemisulfate (0.01% w/v)</td><td></td><td></td><td>FDA (IND 102,217)</td></tr><tr><td>Laser Diode*</td><td>Nichia Corporation</td><td>Not Applicable</td><td>455-nm</td></tr><tr><td>Image Capture*</td><td>Labview</td><td>Not Applicable</td><td></td></tr><tr><td>Spray Catheter</td><td>Olympus</td><td>Not Applicable</td><td></td></tr><tr><td colspan="4">*Equipment specifics within Reference 6. **Equipment specifics within Reference 7</td></tr></tbody></table>

diagnosis of neoplasia in barrett&#8217;s esophagus using vital-dye enhanced fluorescence imaging

从肿瘤在体内分化良性上皮化生Barrett食管（BE）的能力仍然是困难的，因为这两种组织类型可以是平的，并没有什么区别与单纯的白色光成像。其结果是，一个模式，突出腺架构将是有益的在食管远端良性上皮肿瘤鉴别。 VFI是一个使用外源性外用荧光造影剂来界定高档异型增生和癌症良性上皮细胞新技术。具体地，荧光图像提供50至100微米的空间分辨率和视高达2.5厘米的字段，允许内镜形象化腺形态。一旦激发，经典巴雷特化生表现为连续，均匀间隔的腺体和整体均匀的形态;与此相反，肿瘤组织出现拥挤腺体框架完全闭塞。在这里，我们提供的概述仪器仪表和枚举这种新技术的协议。虽然VFI提供了一次胃肠病与可疑组织的腺体架构，细胞不典型增生不能用这种方式解决。因此，人们不能形态学通过该成像方式区分巴雷特化生从与低度不典型增生。通过权衡在与视图更大字段分辨率的降低，该成像系统可用于在至少一个红色标记成像器件对目标和活检可疑病灶;然而，如果精度的措施是令人鼓舞的，VFI可能成为肿瘤的诊断（定义为高档异型增生或癌症）在食管远端的标准成像技术。

图1B描绘了经典的Barrett食管，没有发育不良正常鳞状上皮所包围的边框。与较平坦的鳞状组织，其圆周分布，并用蓝色箭头表示开始，无光泽荧光的均匀区域的存在与无腺体结构。绿色箭头表示周围的鳞状上皮组织的圆形绿线。本大纲是神器从内窥镜的上限所致。移动到位于中心位置的Barrett的组织，腺体结构可以被定义为周围较暗的管腔绿色荧光。虽然一些腺被拉长，有相邻的腺体之间几乎没有失真，如腺体的宽度相似，并且边缘有明确的规定。最后，腺体和流明间隔均匀，无结块或挤占存在。 图2B描绘Barrett食管与低度不典型增生。这是重要的是要注意，尽管有低度异型增生存在，这不能由形态学标准通过这种成像的方式可视化。因此，基于形态模式，这种组织则仍会被界定为良性。而同质腺体和流明的黄色椭圆形的提​​示仅仅是化生，蓝色椭圆表示这主要是联合起来腺体的区域。也就是说，腺体的厚度增加了，而较暗的管腔腔已成为薄且几乎不存在。这些拥挤，轻微扭曲的腺体，但是，有离散，边界和趋于均匀。此外，不存在抹杀形式存在腺体的边缘是平滑的，从而所述组织仍然是良性的。红色箭头表示组织的失焦和视频可以很容易区别开来。最后，黑色箭头所示的气泡是神器。 在图3B中 ，红色椭圆表示与高品位DYS最突出的地区plasia。这些腺体拥挤，因为他们有不规则的薄边框以及组织与腺体结构抹杀附近地区。也就是说，腺不再是独立的，而是融合在一起，与它们的管腔是小型和不规则。虽然小，有些流明的持续存在可能表示高度异型增生，因为它是更经常的浸润性癌，其中管腔完全丧失。与此相反的高档异型增生，黄色椭圆形凸显恶性组织。在这里，腺体结构是抹杀和流明基本上不存在。 图4B示出了一个位于中心的腺癌。首先，请注意红色椭圆内的肿瘤和腺体结构与管腔没有完全闭塞。这种闭塞可比较的癌症由蓝色箭头，其中拥有一些腺体框架指示的组织时，还可以理解。在左边，灰重ctangle突出的鳞状上皮，它可以在视频被更好地理解，当在内窥镜盘在它的全部内容。这鳞状上皮组织是平淡的荧光，无腺体结构的平坦均匀的区域。 <img alt="图1" src="/files/ftp_upload/50992/50992fig1highres.jpg" /> 图1瘤在Barrett食管:白光内镜与活体染料荧光成像。与白光内镜取（左）图像。（右）与相应的重要染料增强荧光成像拍摄的图像。 <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/50992/50992fig1highres.jpg" target="_blank">请点击这里查看这个数字的放大版本。</a>

Watch this Scientific Journal Video about Diagnosis of Neoplasia in Barrettâ€™s Esophagus using Vital-dye Enhanced Fluorescence Imaging at JoVE.com

Diagnosis of Neoplasia in Barrett&#8217;s Esophagus using Vital-dye Enhanced Fluorescence Imaging

重要的染料增强荧光成像（VFI）是生物体内的技术了一种新的，结合高分辨率成像上皮外源外用荧光对比，从而突出腺体形态及食管远端划定瘤（高档异型增生和癌症）。

用生命染增强荧光成像瘤在Barrett食管诊断

The ability to differentiate benign metaplasia in Barrett&#8217;s Esophagus (BE) from neoplasia in vivo remains difficult as both tissue types can be flat ...

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Unit 1

Disorders of the Esophagus

SCIENTISTS IN ACTION

12.0K 次观看.  Icahn School of Medicine at Mount Sinai. 重要的染料增强荧光成像（VFI）是生物体内的技术了一种新的，结合高分辨率成像上皮外源外用荧光对比，从而突出腺体形态及食管远端划定瘤（高档异型增生和癌症）。 

视频: Diagnosis of Neoplasia in Barrett’s Esophagus using Vital-dye Enhanced Fluorescence Imaging

Elastic Staining on Paraffin-embedded Slides of pT3N0M0 Gastric Cancer Tissue

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and eosin (H&amp;E) staining but can be visualized through elastic staining. One of the important benefits of elastic staining is that it makes it easy to determine whether tumor cells have invaded beyond the elastic lamina. This helps in examining the extent of peritoneal surface invasion, thereby distinguishing the pT3 and pT4 stages in gastrointestinal cancer. In this study, we present a protocol to identify elastic fibers in the formalin-fixed, paraffin-embedded pT3N0M0 gastric cancer tissue sections. We prepare 5-&#181;m paraffin sections fixed on slides, and then all the slides are deparaffinated and rehydrated during the staining procedure. Subsequently, all the sections are oxidized by potassium permanganate, bleached by oxalic acid, stained by elastic staining, and counterstained by Van Gieson's. Finally, we examine the effect of staining; elastic lamina is stained blue-black and collagen fibers are stained red, while the cancer cells are stained in varying shades of yellow. We also describe in detail the methods used for determining the positional relationship between cancer cells and elastic lamina. This method is simple, low-cost, and widely applicable in the identification of peritoneal surface invasion in gastrointestinal cancer because of its outstanding selectivity for elastic fibers and authentic results.

Here, we present a protocol of elastic staining to identify elastic fibers in pT3N0M0 gastric cancer tissues on formalin-fixed, paraffin-embedded sections. Subsequently, we describe a method to determine whether tumor cells invade beyond the elastic lamina.

pT3N0M0 胃癌组织石蜡嵌入幻灯片上的弹性染色

The elastic lamina, which is usually located in the sub-mesothelial layer, adjacent to the peritoneum mesothelial cells, is amphiphilic on hematoxylin and ...

科研

癌症研究

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite fluorescent in situ elements through the optical fibers, and then record some of the emitted photons, via the same optical fibers. The light source is a laser that sends the exciting light through an element within the fiber bundle and as it scans over the sample, recreates an image pixel by pixel. As this scan is very fast, by combining it with dedicated image processing software, images in real time with a frequency of 12 frames/sec can be obtained.
We developed a technique to quantitatively characterize capillary morphology and function, using a confocal fluorescence videomicroscopy device. The first step in our experiment was to record 5 sec movies in the four quadrants of the tumor to visualize the capillary network. All movies were processed using software (ImageCell, Mauna Kea Technology, Paris France) that performs an automated segmentation of vessels around a chosen diameter (10 &mu;m in our case). Thus, we could quantify the 'functional capillary density', which is the ratio between the total vessel area and the total area of the image. This parameter was a surrogate marker for microvascular density, usually measured using pathology tools.
The second step was to record movies of the tumor over 20 min to quantify leakage of the macromolecular contrast agent through the capillary wall into the interstitium. By measuring the ratio of signal intensity in the interstitium over that in the vessels, an 'index leakage' was obtained, acting as a surrogate marker for capillary permeability.

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

In this paper, we present a method to analyze tumor microvessels in vivo using dynamic contrast-enhanced fluorescence videomicroscopy. Two quantitative parameters were acquired: functional capillary density reflecting the vascularity of the tumor, and index leakage reflecting the leakiness of the endothelial wall.

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite ...

医学

Sentinel Lymph Node Mapping and Biopsy for Endometrial Cancer at Early Stage with Laparoscopy

Sentinel lymph node (SLN) mapping and biopsy is a promising technique for visualizing and evaluating lymph node status in cancer. This approach has been recommended for low-risk endometrial cancer (EC) patients by authoritative international guidelines, but it has not been performed broadly in China and worldwide. This work aims to describe detailed SLN mapping and biopsy procedures to promote the clinical application. SLN mapping and postoperative pathologic ultrastaging were conducted in a patient with low-risk EC using indocyanine green (ICG) dye to track the SLNs under laparoscopy and resecting them completely for ultrastaging. In conclusion, this protocol describes details of ICG injection, and SLN mapping and biopsy in EC patients based on the experiences gained during clinical practice.

This protocol describes the identification and resection of sentinel lymph nodes to make the operation as easy and minimally invasive as possible.

腹腔镜检查早期子宫内膜癌的前哨淋巴结定位和活检

Sentinel lymph node (SLN) mapping and biopsy is a promising technique for visualizing and evaluating lymph node status in cancer. This approach has been ...

肿瘤微环境的多重免疫荧光和空间图像分析

Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment

The tumor microenvironment (TME) is composed of a plethora of different cell types, such as cytotoxic immune cells and immunomodulatory cells. Depending on its composition and the interactions between cancer cells and peri-tumoral cells, the TME may affect cancer progression. The characterization of tumors and their complex microenvironment could improve the understanding of cancer diseases and may help scientists and clinicians to discover new biomarkers.
We recently developed several multiplex immunofluorescence (mIF) panels based on tyramide signal amplification (TSA) for the characterization of the TME in colorectal cancer, head and neck squamous cell carcinoma, melanoma, and lung cancer. Once the staining and scanning of the corresponding panels are completed, the samples are analyzed on an image analysis software. The spatial position and the staining of each cell are then exported from this quantification software into R. We developed R scripts that allow us not only to analyze the density of each cell type in several tumor compartments (e.g. the center of the tumor, the margin of the tumor, and the stroma) but also to perform distance-based analyses between different cell types.
This particular workflow adds a spatial dimension to the classical density analysis already routinely performed for several markers. mIF analysis could allow scientists to have a better understanding of the complex interaction between cancer cells and the TME and to discover new predictive biomarkers of response to treatments, such as immune checkpoint inhibitors, and targeted therapies.

作者聚焦：多重免疫荧光结合空间图像分析用于肿瘤微环境的临床和生物学评估  

In this article, a protocol for manual tyramide signal amplification (TSA) multiplex immunofluorescence (mIF) combined with image analysis and spatial analysis is described. This protocol can be used with formalin-fixed paraffin-embedded (FFPE) sections for the staining of two to six antigens per slide depending on the slide scanner available in the laboratory.

多重免疫荧光结合空间图像分析，用于肿瘤微环境的临床和生物学评估

The tumor microenvironment (TME) is composed of a plethora of different cell types, such as cytotoxic immune cells and immunomodulatory cells. Depending ...

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

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions preclinically. However, valid assessment of pathological alterations often requires histological analysis, and when performed ex vivo, necessitates death of the animal. Therefore in conventional experimental settings, intra-individual follow-up examinations are rarely possible. Thus, development of murine endoscopy in live mice enables investigators for the first time to both directly visualize the gastrointestinal mucosa and also repeat the procedure to monitor for alterations. Numerous applications for in vivo murine endoscopy exist, including studying intestinal inflammation or wound healing, obtaining mucosal biopsies repeatedly, and to locally administer diagnostic or therapeutic agents using miniature injection catheters. Most recently, molecular imaging has extended diagnostic imaging modalities allowing specific detection of distinct target molecules using specific photoprobes. In conclusion, murine endoscopy has emerged as a novel cutting-edge technology for diagnostic experimental in vivo imaging and may significantly impact on preclinical research in various fields.

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

Small animal imaging techniques allow serial diagnostic examinations and therapeutic interventions in vivo. Recently, the scope of applications has significantly widened and currently includes assessment of colonic tumor development, wound healing and monitoring of inflammation. This protocol illustrates these diverse potential applications of murine endoscopy.

鼠内镜<em&gt;体内</em&gt;发生和肠道的评价多模式成像伤口愈合和炎症

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

Multimodal Imaging and Spectroscopy Fiber-bundle Microendoscopy Platform for Non-invasive, In Vivo Tissue Analysis

Recent fiber-bundle microendoscopy techniques enable non-invasive analysis of in vivo tissue using either imaging techniques or a combination of spectroscopy techniques. Combining imaging and spectroscopy techniques into a single optical probe may provide a more complete analysis of tissue health. In this article, two dissimilar modalities are combined, high-resolution fluorescence microendoscopy imaging and diffuse reflectance spectroscopy, into a single optical probe. High-resolution fluorescence microendoscopy imaging is a technique used to visualize apical tissue micro-architecture and, although mostly a qualitative technique, has demonstrated effective real-time differentiation between neoplastic and non-neoplastic tissue. Diffuse reflectance spectroscopy is a technique which can extract tissue physiological parameters including local hemoglobin concentration, melanin concentration, and oxygen saturation. This article describes the specifications required to construct the fiber-optic probe, how to build the instrumentation, and then demonstrates the technique on in vivo human skin. This work revealed that tissue micro-architecture, specifically apical skin keratinocytes, can be co-registered with its associated physiological parameters. The instrumentation and fiber-bundle probe presented here can be optimized as either a handheld or endoscopically-compatible device for use in a variety of organ systems. Additional clinical research is needed to test the viability of this technique for different epithelial disease states.

Multimodal Imaging and Spectroscopy Fiber-bundle Microendoscopy Platform for Non-invasive, In Vivo Tissue Analysis

The assembly and use of a multimodal microendoscope is described which can co-register superficial tissue image data with tissue physiological parameters including hemoglobin concentration, melanin concentration, and oxygen saturation. This technique can be useful for evaluating tissue structure and perfusion, and can be optimized for individual needs of the investigator.

多峰成像和光谱学纤维束显微内窥镜纲要非侵入性，<em&gt;在体内</em&gt;组织分析

Recent fiber-bundle microendoscopy techniques enable non-invasive analysis of in vivo tissue using either imaging techniques or a combination of ...

生物工程

胃低级别上皮内瘤变风险预测模型

Establishment and Evaluation of a Risk Prediction Model for Pathological Escalation of Gastric Low-Grade Intraepithelial Neoplasia

The study aims to explore the risk factors for pathological escalation after endoscopic surgery for gastric low-grade intraepithelial neoplasia (LGIN) and to establish and evaluate a risk prediction model for LGIN. A total of 120 patients diagnosed with gastric LGIN by biopsy and endoscopic submucosal dissection (ESD) between November 2020 and June 2022 were retrospectively analyzed. Gender, age, Helicobacter pylori (HP) infection, lesion size, lesion location, morphology, gastric mucosal congestion, nodules status, surface ulceration and erosion, and ME-observation of all patients were collected and divided into upgraded and non-upgraded groups according to the biopsy and ESD postoperative pathological diagnosis results. Independent risk factors for pathological escalation after ESD surgical treatment were screened by logistic regression analysis, and a risk prediction model was established. Among the 120 patients with gastric LGIN, 49 patients developed postoperative pathological upgrading; the rate of pathological upgrading was 40.83%. Among them, 42 patients were upgraded to high-grade intraepithelial neoplasia (HGIN), 1 case was upgraded to advanced gastric cancer, and 6 cases were upgraded to early gastric carcinoma (EGC). Univariate analysis showed that age, lesion size, gastric mucosal congestion, surface ulcers, and erosion were significantly different between the groups (p &lt; 0.05). Multivariate Logistic regression analysis revealed that age &#8805;60 years, focal length &#8805;2 cm, gastric mucosal congestion, and surface ulceration and erosion were independent risk factors for postoperative pathological escalation in patients with gastric LGIN. Final joint probability forecasting model for P = 1/[1 + e(26.515-0.161 x &#946;1-0.357 x &#946;2+0.039 x &#946;3-0.269 x &#946;4)]. Age, lesion size &#8805;2 cm, gastric mucosal congestion, and lesion surface ulceration and erosion are risk factors for postoperative pathological upgrading in patients with gastric LGIN. The risk prediction model established in this study based on risk factors has predictive value and can provide a scientific reference for the clinical treatment of patients with gastric LGIN.

作者聚焦：推进胃肠道肿瘤的早期检测和治疗

Here, we performed a systematic evaluation of patients diagnosed with gastric low-grade intraepithelial neoplasia by previous endoscopic biopsy and obtained a pathological diagnosis by complete resection of the lesion by endoscopic submucosal dissection (ESD), analyzing factors that potentially increase the risk of pathological escalation.

胃低级别上皮内瘤变病理升级风险预测模型的建立与评价

The study aims to explore the risk factors for pathological escalation after endoscopic surgery for gastric low-grade intraepithelial neoplasia (LGIN) and ...

用生命染增强荧光成像瘤在Barrett食管诊断

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此视频中的章节

pT3N0M0 胃癌组织石蜡嵌入幻灯片上的弹性染色

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

腹腔镜检查早期子宫内膜癌的前哨淋巴结定位和活检

多重免疫荧光结合空间图像分析，用于肿瘤微环境的临床和生物学评估

多重免疫荧光结合空间图像分析，用于肿瘤微环境的临床和生物学评估

多重免疫荧光结合空间图像分析，用于肿瘤微环境的临床和生物学评估

鼠内镜

多峰成像和光谱学纤维束显微内窥镜纲要非侵入性，

胃低级别上皮内瘤变病理升级风险预测模型的建立与评价

胃低级别上皮内瘤变病理升级风险预测模型的建立与评价