Name: spatial measurements of perfusion, interstitial fluid pressure and liposomes accumulation in solid tumors
Uploaded: 2016-08-18T13:00:00.000+00:00
Duration: 9 min
Description: Watch this Scientific Journal Video about Spatial Measurements of Perfusion, Interstitial Fluid Pressure and Liposomes Accumulation in Solid Tumors at JoVE.com

The authors would like to thank Dr. Javed Mahmood for assistance with culturing MDA-MB-231 cells and implanting the MDA-MB-231 xenografts, Linyu Fan for preparing the CT-liposomes. Shawn Stapleton is grateful for funding from the Natural Sciences and Engineering Research Postgraduate Scholarships Program and the Terry Fox Foundation Strategic Initiative for Excellence in Radiation Research for the 21st Century (EIRR21) at CIHR. This study was supported by grants from the Terry Fox New Frontiers Program (020005) and the Canadian Institutes of Health Research (102569).

所有的体内实验是由大学健康网络机构动物护理和使用委员会批准的方案下进行的。 1.动物模型<ol><li> 5至7×10 6个 MDA-MB-231乳腺癌肿瘤细胞在含10％胎牛血清（FBS）和100倍稀释青霉素-链霉素的一起之间培养。 </li><li>收获细胞时，他们使用的是0.05％胰蛋白酶EDTA溶液80％汇合。 3-5分钟后中和胰蛋白酶EDTA的DMEM的3倍量。取细胞的15微升等分，并使用血球计数。离心细胞向在200×g离心5分钟沉淀，并在HBS以每毫升10×10 6个细胞的浓度重悬。 </li><li>植入皮下（SC）由每个8至12周大的雌性SCID小鼠的后肢注射1到2×10 6个细胞瘤（N = 5）。使用标准的25针摹注射。 </li><li>监视器ŧumor用卡尺增长（体积= 0.5×长×宽2），并开始进行测量，一旦SC肿瘤已经达到了量&gt; 200 立方毫米（约7-9天）。 </li></ol> 2. CT-脂质体的制备与表征<ol><li>脂质体制剂<ol><li>溶解脂质组分（200毫摩尔/ L），用于对CT-脂质体，其中包括1,2-二棕榈酰-sn-甘油基-3-磷酸胆碱（DPPC），胆固醇（CH），和1,2-二硬脂酰-sn-甘油-3- -phosphoethanolamine-N - 聚（乙二醇）2000（DSPE-PEG2000）的无水乙醇在70℃下以55的摩尔比:40:5的DPPC CH:DSPE-PEG2000。 </li><li>通过在70℃下保持热蒸发乙醇，然后加入CT造影剂碘海醇（300毫克/毫升碘）到该溶液中，使得最终的脂质浓度为100mM。 </li><li>保持在70℃4小时频繁涡旋该溶液中。 </li><li>为了获得单层囊，挤出样品5恬通过两个ES，在250 psi的压力层叠200nm的孔尺寸的膜，并再次通过5个循环通过两个层叠80nm的孔径的膜，在400磅使用10ml的脂质挤出机挤出。吸取10毫升脂质体的体积到挤压机在每个挤压周期的开始，并收集到每个挤压周期后的无菌锥形管或玻璃管形瓶中。 </li><li>使用100 kDa的分子量切断对250倍体积过量的0.02毫米的HEPES缓冲的盐水溶液（HBS，pH值7.4）（MWC）透析袋透析16小时除去未包封碘海醇。例如，地点1毫升的透析袋用250毫升HBS的外袋在烧杯内的脂质体溶液。 </li><li>集中根据制造商的说明使用75万nomical MWC商业切向流系统对CT-脂质体。集中到约55毫克ml的最终碘浓度-1。 </li></ol></li><li>脂质体表征<ol><li>通过使用10倍体积的过量的乙醇以释放ioxehol然后稀释用100倍体积的过量的去离子水破裂对CT-脂质体测量包封率（ 即 10微升脂质体的使用的乙醇100微升，然后稀释破裂至10毫升的最终体积）。 </li><li>使用带有检测的UV光谱仪上在245纳米的波长确定碘海醇的浓度。通过采取释放碘海醇剂的比例量剂的量计算出的封装效率制备过程中加入。 </li><li>使用根据制造商的说明，动态光散射粒度分析仪系统测量的流体动力学直径和zeta电位。稀释由200倍（ 即 5微升脂质体在1ml最终体积的）对CT-脂质体溶液在去离子水中，以促进测量。 </li></ol></li></ol> 3.肿瘤微循环和CT-脂质体的CT成像分配注:按照制造商的说明，如果使用不同的软件版本或设备执行体积扫描。 <ol><li>麻醉，用2％异氟烷以医用空气或氧气混合各小鼠和按捏脚趾和观察没有反应确认。适用软膏的眼睛，防止干燥时的麻醉下。用胶带爪子薄塑料板固定在俯卧的动物。 </li><li>将一个自定义的27克导管，连接到PE10管20厘米，到侧尾静脉并在插好几块胶带固定。 </li><li>制备1ml注射器包含对CT-脂质体中的至少200微升。制备1ml注射器用生理盐水使用冲洗导管。最后，制备1ml注射器具有至少150微升游离碘海醇与盐水混合（9:1体积比）。 </li><li>把鼠标放在微CT扫描仪床上俯卧。用激光定位系统以放置致瘤R IN大约每个扫描相同的方向。 </li><li>放置在CT脂质体注射器在注射器泵和导管连接到注射器。设置的每秒10微升泵速率。 </li><li>通过使用CT扫描仪控制台软件进行明暗校准扫描初始化系统。选择每个感兴趣的成像协议的亮暗扫描选项，从下拉菜单中亮暗，然后按下扫描按钮开始校准。 </li><li>执行前的任何造影剂注射的肿瘤的体积解剖显微CT。再看CT扫描仪控制台软件指标，以确保CT扫描仪的安全联锁装置已经被清除。在CT扫描器控制台选择扫描选择80千伏70毫安的管电流的X射线能量，并且随着时间的推移16秒捕获1000图像的预测。按下扫描按钮开始扫描。 </li><li>使用注射泵在400毫克碘kg的浓度注入CT-脂质体的大丸剂-1。设置泵注入大约150微升（假设将25g小鼠）的体积。按下泵注入"开始"按钮。手动冲洗导管与盐水的50微升（两次导管的体积），以确保在注射整个剂量和导管是明确的。 </li><li>等待10分钟的CT-脂质体的注射后，然后执行使用在3.5中描述的相同的方法和设置的第二解剖扫描。 </li><li>使用在3.3中描述的相同的喷射率设定:（1体积比9）通过设置在注射器泵注入100μl的游离碘海醇与盐水混合的体积执行一个DCE-CT扫描。 <ol><li>于CT扫描器控制台选择使用80千伏，90毫安的管能量的x射线能量设定，并捕获416图像投影每一秒用于第一30秒和随后的获取每10秒5分钟动态扫描。捕获5秒DCE-CT数据，然后按下该注射液的启动按钮ñ泵。 </li><li>在DCE-CT扫描后进行解剖体积微型CT扫描。 </li></ol></li><li>捕获解剖CT图像的CT-脂质体的48和72小时后喷射之间，使用相同的立体CT设置在3.5步骤所述。 </li><li>重建使用GPU-重建软件解剖CT和DCE-CT数据。 <ol><li>加载图像进重建软件。选择感兴趣的区域，以通过使用鼠标绘制的ROI在图像重建。设置重建成像的保存位置和文件名，选择输出文件类型为".MAT"。 注:该软件会自动设置重建像素大小为0.153 0.153点¯x0.153点¯xmm 3的对解剖扫描和 0.153 0.153点¯x点¯x0.462毫米3 DCE-CT扫描。点击"开始重建"按钮。 </li></ol></li><li>使用CT-脂质体的预喷射和10分钟后注射扫描来计算血浆体积分数如前所述3。此外，使用预喷射和碘海醇的5分钟注射后扫描如前所述7来计算间隙体积分数。 </li><li>通过导入DCE-CT数据转换为提供给肿瘤体积内找出感兴趣区域（ROI）的区域能力的软件获得时间强度曲线（TICS资本流动）。然后计算在ROI的平均CT增强作为时间的函数。在这个实验中的定制软件的开发是为了确定一个投资回报率和计算TIC。 </li><li>通过使用二室示踪剂动力学模型拟合测定国际信托获得灌注和血管通透性的定量估计。配件可使用的DCE-CT分析软件来执行，并使用等离子体体积分数的先验估计和插页体积分数为在两隔室示踪剂动力学模型的固定参数。使用先前报道的方法来获取血浆的先验估计一个和间质体积分数14。 </li></ol> 4.肿瘤组织间液压力的空间测量<ol><li>为了测量IFP通过50厘米PE20聚乙烯管的地下25脊椎针头连接到压力换能器和对IFP采集系统。冲洗整个系统用硫酸肝素/盐水溶液（1:10）。用消毒后再使用70％的异丙醇针。 </li><li>打开采集系统，并启动IFP采集软件和加载设置文件到系统收​​购毫米汞柱IFP测量校准。点击获取按钮连续收集数据IFP。 </li><li>执行的CT-脂质体的48和72小时后喷射之间IFP测量（这对应于肿瘤的CT-脂质体的峰积累的大致时间），使用在4.8中描述的方法。附加IFP针对CT-IFP机器人。 </li><li>执行校准扫描对准的坐标系对CT-IFP机器人和CT扫描仪。添加基准标记附着到CT-IFP机器人和在四个不同位置的基准标记进行的四立体CT扫描。 <ol><li>启动CT-IFP机器人控制器软件，初始化机器人，并通过输入X，Y，Z目标位置，然后点击"进入"按钮，机器人移动到三个位置。 </li><li>以CT扫描在下面的X，Y，Z坐标:（1）0,0,0; （2）-10,0,0; （3）0,7,0;和（4）0,0,10。选择一个90千伏为10 mA，16使用CT扫描仪软件，然后按"开始"开始扫描秒扫描。在3.10中描述重建扫描。 </li></ol></li><li>启动CT-IFP机器人比对软件。点击"注册数据"区域中加载的"添加"按钮，然后在4.3得到的4个重建登记扫描，然后点击"打开"。 注:基准标记的像素位置将被自动输入到softwa回覆。 <ol><li>点击"计算转换"按钮，然后点击"应用转换"按钮。这产生将被用于转换对CT-IFP机器人坐标系到CT扫描器的坐标系对准数据。校准完成后，贴上动物平台，以CT-IFP机器人。 </li></ol></li><li>麻醉，用2％异氟烷以医用空气或氧气混合各小鼠和按捏脚趾和观察没有反应确认。固定对CT-IFP机器人平台对动物和定位鼠标，使得肿瘤的CT-IFP机器人系统进​​行访问。固定用胶带，使得它不IFP针插入期间移动至肿瘤。 </li><li>执行前，插入针IFP解剖显微CT扫描。重建用3.10描述的步骤的CT数据。 </li><li>加载针预先插入CT数据到对CT-IFP机器人对准软件。调整窗口和水平可视化的肿瘤。点击次在任何图像肿瘤电子边缘，然后在第二个边缘的位置。 注:该软件将计算出的一系列位置的沿在两个位置之间的直线。注意x，y和一系列从列表5-8均匀间隔的位置中的Z坐标。 </li><li>冲洗以插入之前肝素盐溶液针准备IFP系统。 </li><li>进入第一预确定针的位置在X，Y，Z，进入对CT-IFP机器人控制软件和压移动到"走"按钮，将机器人移动到所需的位置。点击"插入针"按钮，插入针头进入组织。 <ol><li>插入针确保IFP针和捏和释放PE20管组织之间良好的流体连通，并指出，IFP测量的增加和返回的IFP采集软件到预先捏值之后。拒绝不回到基线测量。 </li></ol></li><li>获得一个插入针解剖CT扫描，然后点击CT-IFP机器人控制软件的"回退针"按钮，退离组织的针。拒绝任何IFP测量其中的IFP值不针撤出后返回到针预先插入值。这意味着针可能在测量过程中已被堵塞。重复步骤对每个针位4.8〜4.10。 </li><li>通过计算相对于在针的插入后的容积CT扫描确定的肿瘤体积的质量中心针端口的x，y和z位置确定肿瘤体积内的针的位置。 </li><li>返回动物的笼子里所有的测量完成后。不要让动物无人值守，并注意观察他们，直到意识已恢复，他们能够保持胸骨斜卧。 </li></ol>

<ol>
	<li>Seo, J. W., Zhang, H., Kukis, D. L., Meares, C. F., Ferrara, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+to+label+preformed+liposomes+with+64Cu+for+positron+emission+tomography+(PET)+imaging.">A novel method to label preformed liposomes with 64Cu for positron emission tomography (PET) imaging.</a> Bioconjugate chemistry. 19 (12), 2577-2584 (2008).</li><li>Huang, H., Dunne, M., Lo, J., Jaffray, D., Allen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+Computed+Tomography-+and+Optical+Image-Based+Assessment+of+Liposome+Distribution.">Comparison of Computed Tomography- and Optical Image-Based Assessment of Liposome Distribution.</a> Molecular Imaging. 12 (3), 148-160 (2013).</li><li>Stapleton, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mathematical+model+of+the+enhanced+permeability+and+retention+effect+for+liposome+transport+in+solid+tumors.">A mathematical model of the enhanced permeability and retention effect for liposome transport in solid tumors.</a> PloS one. 8 (12), e81157 (2013).</li><li>Zheng, 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=A+multimodal+nano+agent+for+image-guided+cancer+surgery.">A multimodal nano agent for image-guided cancer surgery.</a> Biomaterials. 67, 160-168 (2015).</li><li>Zheng, J., Liu, J., Dunne, M., Jaffray, D. A., Allen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+performance+of+a+liposomal+vascular+contrast+agent+for+CT+and+MR-based+image+guidance+applications.">In vivo performance of a liposomal vascular contrast agent for CT and MR-based image guidance applications.</a> Pharmaceutical research. 24 (6), 1193-1201 (2007).</li><li>Harrington, K. 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=Effective+targeting+of+solid+tumors+in+patients+with+locally+advanced+cancers+by+radiolabeled+pegylated+liposomes.">Effective targeting of solid tumors in patients with locally advanced cancers by radiolabeled pegylated liposomes.</a> Clinical Cancer Research. 7 (2), 243-254 (2001).</li><li>Stapleton, S., Allen, C., Pintilie, M., Jaffray, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+perfusion+imaging+predicts+the+intra-tumoral+accumulation+of+liposomes.">Tumor perfusion imaging predicts the intra-tumoral accumulation of liposomes.</a> J Control Release. 172 (1), 351-357 (2013).</li><li>Lammers, T., Kiessling, F., Hennink, W. E., Storm, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanotheranostics+and+image-guided+drug+delivery:+current+concepts+and+future+directions.">Nanotheranostics and image-guided drug delivery: current concepts and future directions.</a> Mol. Pharm. 7, 1899-1912 (2010).</li><li>Stapleton, S., Milosevic, M. 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> Cancer Targeted Drug Delivery. , 241-272 (2013).</li><li>Blanco, E., Shen, H., Ferrari, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+of+nanoparticle+design+for+overcoming+biological+barriers+to+drug+delivery.">Principles of nanoparticle design for overcoming biological barriers to drug delivery.</a> Nature biotechnology. 33 (9), 941-951 (2015).</li><li>Heldin, C. H., Rubin, K., Pietras, K., Ostman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+interstitial+fluid+pressure+-+an+obstacle+in+cancer+therapy.">High interstitial fluid pressure - an obstacle in cancer therapy.</a> Nat Rev Cancer. 4 (10), 806-813 (2004).</li><li>Chauhan, V. P., Stylianopoulos, T., Boucher, Y., Jain, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delivery+of+molecular+and+nanoscale+medicine+to+tumors:+transport+barriers+and+strategies.">Delivery of molecular and nanoscale medicine to tumors: transport barriers and strategies.</a> Annual review of chemical and biomolecular engineering. 2, 281-298 (2011).</li><li>Bax, J. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=3D+image-guided+robotic+needle+positioning+system+for+small+animal+interventions.">3D image-guided robotic needle positioning system for small animal interventions.</a> Medical physics. 40 (1), 011909 (2013).</li><li>Stapleton, S., Milosevic, M., Tannock, I. F., Allen, C., Jaffray, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+intra-tumoral+relationship+between+microcirculation,+interstitial+fluid+pressure+and+liposome+accumulation.">The intra-tumoral relationship between microcirculation, interstitial fluid pressure and liposome accumulation.</a> Journal of Controlled Release. 211, 163-170 (2015).</li><li>Stapleton, S., Allen, C., Pintilie, M., Jaffray, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+perfusion+imaging+predicts+the+intra-tumoral+accumulation+of+liposomes.">Tumor perfusion imaging predicts the intra-tumoral accumulation of liposomes.</a> J Control Release. 172 (1), 351-357 (2013).</li><li>Brix, G., Zwick, S., Kiessling, F., Griebel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacokinetic+analysis+of+tissue+microcirculation+using+nested+models:+multimodel+inference+and+parameter+identifiability.">Pharmacokinetic analysis of tissue microcirculation using nested models: multimodel inference and parameter identifiability.</a> Medical physics. 36 (7), 2923-2933 (2009).</li><li>Brix, G., Griebel, J., Kiessling, F., Wenz, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracer+kinetic+modelling+of+tumour+angiogenesis+based+on+dynamic+contrast-enhanced+CT+and+MRI+measurements.">Tracer kinetic modelling of tumour angiogenesis based on dynamic contrast-enhanced CT and MRI measurements.</a> European journal of nuclear medicine and molecular imaging. 37 (1), 30-51 (2010).</li></ol>

The authors have nothing to disclose

本文中所呈现用于基于图像的测量方法能够肿瘤微循环特性，IFP，和CT-脂质积累的空间分布的确定。以前的尝试涉及这些属性都依赖于执行在多个肿瘤携带动物散装测量，因此缺乏灵敏度阐明负责该已普遍被观察到纳米尺寸的药物输送系统15在肿瘤内积聚的异质性的机制。 DCE-CT提供了测量在肿瘤微循环特性的肿瘤内变化的工具，立体CT提供的CT-脂质沉积动力学的准确描绘，与CT-IFP机器人系...

测量纳米颗粒的药物递送系统的肿瘤内积累可以提供，以确定是否细胞毒性药物的适当浓度已在肿瘤内实现的​​一个重要工具。 "图象能够"脂质体系统的发展允许非侵入性和定量的体内检测用成像方式如正电子发射断层扫描（PET）1，光学荧光2，和计算机断层扫描（CT）3的药物递送载体的， 4和磁共振成像（MRI）5。成像已被用于确定药代动力学和脂质体递送系统的生物分布和以显示跨学科和肿瘤内的异质性在纳米颗粒累积6,7的程度。然而，纳米粒子成像本身并不能识别已经给他们可怜的积累和分配贡献了生物屏障。这方面的知识是最重要的，将r更有效的配方憩的发展和战略，以提高肿瘤内积聚8。它已被证实的治疗策略可以应用于调节从而提高纳米颗粒运输9特定生物屏障。此外，纳米颗粒制剂已经开发了专门克服特定生物传输阻挡10。在这两种情况下，生物屏障的测量可以被用来指导使用合适的纳米颗粒的药物递送策略的。 肿瘤微循环和升高IFP被认为是纳米颗粒的肿瘤内蓄积的两个关键因素，如脂质体，在实体瘤9,11。然而，有助于脂质体积累穷人服务的其他障碍包括致密的细胞外基质，防渗血管和实体组织的压力12。这些障碍在一个时空相关方式，与异常血流和升高的组织间隙液压作为驱动纳米颗粒的初始传送和外渗的两个重要因素。如先前所讨论的，建立肿瘤微循环，升高的IFP和脂质体的肿瘤内蓄积之间的关系是当务之急脂质体的成像数据的适当解释。在此定量的方法来衡量实体瘤，肿瘤微循环，IFP升高和积累纳米之间的关系呈现。这是通过使用容积CT成像，肿瘤微循环使用动态对比增强计算机断层成像执行的CT脂质体的造影剂的肿瘤内分布的共定位测量完成，使用图像引导的机器人针定位系统，肿瘤IFP称为对CT-IFP机器人13。

<table><tbody><tr><td>MDA-MB-231 metastatic breast adenocarcinoma tumor cells&amp;nbsp;</td><td>ATCC</td><td>HTB-26</td><td></td></tr><tr><td>Dulbecco&#039;s Modified Eagle Medium (DMEM)&amp;nbsp;</td><td>Life Technologies</td><td>11965-092</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Sigma-Aldrich</td><td>F1051</td><td></td></tr><tr><td>HyClone Penicillin-Streptomycin 100x Solution</td><td>GE Healthcare Life Sciences</td><td>SV30010</td><td></td></tr><tr><td>Trypsin-EDTA (0.05%), phenol red</td><td>ThermoFisher Scientific</td><td>25300-054</td><td></td></tr><tr><td>1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)</td><td>Avanti Lipids Inc., USA</td><td>850355P</td><td></td></tr><tr><td>Cholesterol (CH)</td><td>Avanti Lipids Inc., USA</td><td>700000P</td><td></td></tr><tr><td>1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-poly(ethylene glycol) 2000 (DSPE-PEG2000)</td><td>Avanti Lipids Inc., USA</td><td>880128P</td><td></td></tr><tr><td>Omnipaque (Iohexol) 300 mg of iodine/ml&amp;nbsp;</td><td>GE Healthcare, CA</td><td></td><td></td></tr><tr><td>80 nm pore size Track-Etch polycarbonate membranes</td><td>Whatman Inc., USA</td><td></td><td></td></tr><tr><td>200 nm pore size Track-Etch polycarbonate membranes</td><td>Whatman Inc., USA</td><td></td><td></td></tr><tr><td>10 m Lipex Extruder&amp;nbsp;</td><td>Nothern Lipids Inc, CA</td><td></td><td></td></tr><tr><td>Dialysis Bag Molecular Weight Cut Off (MWCO) of 8 kDa</td><td>Spectrum Labs, USA&amp;nbsp;</td><td></td><td></td></tr><tr><td>750,000 Nomical Molecular Weight Cut Off (NMWC) Tangential flow column&amp;nbsp;</td><td>MidGee ultrafiltration cartridge, GE Healthcare, CA</td><td></td><td></td></tr><tr><td>Peristaltic pump&amp;nbsp;</td><td>Watson Marlow Inc., USA</td><td></td><td></td></tr><tr><td>UV spectrometer</td><td>Helios &amp;gamma;, Spectronic Unicam,&amp;nbsp; USA</td><td></td><td></td></tr><tr><td>90Plus particle size analyzer&amp;nbsp;</td><td>Brookhaven, Holtsville, USA</td><td></td><td></td></tr><tr><td>eXplore Locus Ultra micro-CT system&amp;nbsp;</td><td>GE Healthcare, CA</td><td></td><td>Manipulated using CT-Console Software</td></tr><tr><td>AxRecon GPU-based Reconstruction</td><td>&amp;nbsp;Acceleware Corp. CA</td><td></td><td></td></tr><tr><td>27 G Catheter SURFLO Winged Infusion Set</td><td>Terumo Medical Products, USA</td><td>SV*27EL</td><td></td></tr><tr><td>PE20 polyethylyne tubing</td><td>Becton Dickinson, USA</td><td>427406</td><td></td></tr><tr><td>Pen tip 25 G &amp;times; 3.5&amp;prime;&amp;prime; Whitacre spinal needle&amp;nbsp;</td><td>Becton Dickinson, USA</td><td>405140</td><td>IFP needle</td></tr><tr><td>P23XL&amp;nbsp; pressure transducer&amp;nbsp;</td><td>Harvard Apparatus, CA</td><td>P23XL</td><td></td></tr><tr><td>PowerLab 4/35, Bridge Amp, with LabChart Pro 7.0</td><td>ADInstruments Pty Ltd., USA</td><td>PL3504, FE221</td><td>IFP acquisition system and acquisition software</td></tr><tr><td>CT-Sabre Small Animall Intervention system (CT-IFP Robot)</td><td>Parallax Innovations, CA</td><td></td><td>Manipulated using CT-IFP robot Control Software</td></tr><tr><td>CT-IFP robot alignment software</td><td></td><td></td><td>Custom Matlab software</td></tr><tr><td>DCE-CT Analysis Software</td><td></td><td></td><td>Custom Matlab software</td></tr><tr><td>Matlab 2013b</td><td>Mathworks, USA</td><td></td><td></td></tr></tbody></table>

spatial measurements of perfusion, interstitial fluid pressure and liposomes accumulation in solid tumors

脂质体的异质肿瘤内积累是其功效的关键决定因素。既混沌肿瘤微循环和高架IFP被链接到基于纳米技术的药物递送系统的异质肿瘤内分布，如脂质体。在本研究中，肿瘤微循环，升高的IFP，和纳米颗粒的积累之间的关系，通过体内实验进行了研究。这是通过使用动态对比增强计算机断层扫描（DCE-CT）和使用连接到微CT扫描仪的新型图像引导的机器人针放置系统肿瘤IFP的测量肿瘤微循环的评估来实现的。脂质体的肿瘤内积聚是由纳米颗粒脂质体制剂稳定地包封造影剂碘海醇（CT-脂质体）的基于图像的CT评价来确定。 CT成像允许的空间分布的共定位肿瘤血流动力学，IFP和CT-脂质体蓄积在乳腺癌的个体皮下异种移植物小鼠模型。测量导致发现该灌注和血​​浆体积分数是脂质体的肿瘤内分布的强介体。此外，结果表明，IFP起着通过调节血流介导的脂质体分布的间接作用。

上述协议应产生的CT-脂质体与碘海醇的包封浓度，平均脂质体直径，和55毫克毫升-1ζ电位，分别91.8±0.3 nm和-45.5±2.5毫伏， 图1a包括代表DCE-CT成像结果，产生一个时间序列，显示在碘海醇的肿瘤内蓄积的经时变化的体积数据。选择肿瘤内的投资回报率产生可使用示踪动力学建模方法以获得灌注，血管通透性，血浆体积分数，和间质体积分数（ 图1b...

Watch this Scientific Journal Video about Spatial Measurements of Perfusion, Interstitial Fluid Pressure and Liposomes Accumulation in Solid Tumors at JoVE.com

Spatial Measurements of Perfusion, Interstitial Fluid Pressure and Liposomes Accumulation in Solid Tumors

The heterogeneous intra-tumoral accumulation of liposomes has been linked to an abnormal tumor microenvironment. Herein methods are presented to measure tumor microcirculation by perfusion imaging and elevated interstitial fluid pressure (IFP) using an image-guided robotic system. Measurements are compared to the intra-tumoral accumulation of liposomes, determined using volumetric micro-CT imaging.

灌注的空间测量，实体肿瘤间质流体压力和脂质体积累

The heterogeneous intra-tumoral accumulation of liposomes is a critical determinant of their efficacy. Both the chaotic tumor microcirculation and ...

spatial-measurements-perfusion-interstitial-fluid-pressure-liposomes

Research

JoVE Journal

Medicine

7.6K 次观看.  University of Toronto. The heterogeneous intra-tumoral accumulation of liposomes has been linked to an abnormal tumor microenvironment. Herein methods are presented to measure tumor microcirculation by perfusion imaging and elevated interstitial fluid pressure (IFP) using an image-guided robotic system. Measurements are compared to the intra-tumoral accumulation of liposomes, determined using volumetric micro-CT imaging.

视频: Spatial Measurements of Perfusion, Interstitial Fluid Pressure and Liposomes Accumulation in Solid Tumors

Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the microvasculature are controlled by varying ultrasound pulsing parameters.  Specifically, we are testing whether such approaches may be used to treat malignant brain tumors through drug delivery and microvascular ablation.  Preliminary studies have been performed to determine whether targeted drug-bearing nanoparticle delivery can be facilitated by the ultrasound mediated destruction of "composite" delivery agents comprised of 100nm poly(lactide-co-glycolide) (PLAGA) nanoparticles that are adhered to albumin shelled microbubbles. We denote these agents as microbubble-nanoparticle composite agents (MNCAs). When targeted to subcutaneous C6 gliomas with ultrasound, we observed an immediate 4.6-fold increase in nanoparticle delivery in MNCA treated tumors over tumors treated with microbubbles co-administered with nanoparticles and a 8.5 fold increase over non-treated tumors. Furthermore, in many cancer applications, we believe it may be desirable to perform targeted drug delivery in conjunction with ablation of the tumor microcirculation, which will lead to tumor hypoxia and apoptosis. To this end, we have tested the efficacy of non-theramal cavitation-induced microvascular ablation, showing that this approach elicits tumor perfusion reduction, apoptosis, significant growth inhibition, and necrosis. Taken together, these results indicate that our ultrasound-targeted approach has the potential to increase therapeutic efficiency by creating tumor necrosis through microvascular ablation and/or simultaneously enhancing the drug payload in gliomas.

Insonation of microbubbles is a promising strategy for tumor ablation at reduced time-averaged acoustic powers, as well as for the targeted delivery of therapeutics. The purpose of the present study is to develop low duty cycle ultrasound pulsing strategies and nanocarriers to maximize non-thermal microvascular ablation and payload delivery to subcutaneous C6 gliomas.

超声造影有针对性的治疗胶质瘤的小鼠通过药物轴承纳米粒子交付和微血管消融

We are developing minimally-invasive contrast agent microbubble based therapeutic approaches in which the permeabilization and/or ablation of the ...

科研

生物工程

Magnetic Resonance Imaging Quantification of Pulmonary Perfusion using Calibrated Arterial Spin Labeling

This demonstrates a MR imaging method to measure the spatial distribution of pulmonary blood flow in healthy subjects 
during normoxia (inspired O2, fraction (FIO2) = 0.21) hypoxia (FIO2 = 0.125), and hyperoxia 
(FIO2 = 1.00). In addition, the physiological responses of the subject are monitored in the MR scan environment. MR images 
were obtained on a 1.5 T GE MRI scanner during a breath hold from a sagittal slice in the right lung at functional residual capacity. An arterial 
spin labeling sequence (ASL-FAIRER) was used to measure the spatial distribution of pulmonary blood flow 1,2 and a multi-echo fast 
gradient echo (mGRE) sequence 3 was used to quantify the regional proton (i.e. H2O) density, allowing the quantification 
of density-normalized perfusion for each voxel (milliliters blood per minute per gram lung tissue). 
With a pneumatic switching valve and facemask equipped with a 2-way non-rebreathing valve, different oxygen concentrations 
were introduced to the subject in the MR scanner through the inspired gas tubing. A metabolic cart collected expiratory gas via expiratory tubing. Mixed expiratory O2 and CO2 concentrations, oxygen consumption, carbon dioxide production, respiratory exchange ratio, 
respiratory frequency and tidal volume were measured. Heart rate and oxygen saturation were monitored using pulse-oximetry. 
Data obtained from a normal subject showed that, as expected, heart rate was higher in hypoxia (60 bpm) than during normoxia (51) or hyperoxia (50) and the arterial oxygen saturation (SpO2) was reduced during hypoxia to 86%. Mean ventilation was 8.31 L/min BTPS during hypoxia, 7.04 L/min during normoxia, and 6.64 L/min during hyperoxia. Tidal volume was 0.76 L during hypoxia, 0.69 L during normoxia, and 0.67 L during hyperoxia. 
Representative quantified ASL data showed that the mean density normalized perfusion was 8.86 ml/min/g during hypoxia, 8.26 ml/min/g during normoxia and 8.46 ml/min/g during hyperoxia, respectively. In this subject, the relative dispersion4, an index of global heterogeneity, was increased in hypoxia (1.07 during hypoxia, 0.85 during normoxia, and 0.87 during hyperoxia) while the fractal dimension (Ds), another index of heterogeneity reflecting vascular branching structure, was unchanged (1.24 during hypoxia, 1.26 during normoxia, and 1.26 during hyperoxia). 
Overview. This protocol will demonstrate the acquisition of data to measure the distribution of pulmonary perfusion noninvasively under conditions of normoxia, hypoxia, and hyperoxia using a magnetic resonance imaging technique known as arterial spin labeling (ASL). 
Rationale: Measurement of pulmonary blood flow and lung proton density using MR technique offers high spatial resolution images which can be quantified and the ability to perform repeated measurements under several different physiological conditions. In human studies, PET, SPECT, and CT are commonly used as the alternative techniques. However, these techniques involve exposure to ionizing radiation, and thus are not suitable for repeated measurements in human subjects.

A MR imaging method to study the distribution of pulmonary blood flow under a variety of physiological conditions, in this case exposure to three different inspired oxygen concentrations: hypoxia, normoxia, and hyperoxia, is described. This technique utilizes human pulmonary physiology research techniques in an MR scanning environment.

使用校准动脉自旋标记磁共振肺灌注显像定量

This demonstrates a MR imaging method to measure the spatial distribution of pulmonary blood flow in healthy subjects 
during normoxia (inspired O2, ...

医学

Assessing Tumor Microenvironment of Metastasis Doorway-Mediated Vascular Permeability Associated with Cancer Cell Dissemination using Intravital Imaging and Fixed Tissue Analysis

The most common cause of cancer related mortality is metastasis, a process that requires dissemination of cancer cells from the primary tumor to secondary sites. Recently, we established that cancer cell dissemination in primary breast cancer and at metastatic sites in the lung occurs only at doorways called Tumor MicroEnvironment of Metastasis (TMEM). TMEM doorway number is prognostic for distant recurrence of metastatic disease in breast cancer patients. TMEM doorways are composed of a cancer cell which over-expresses the actin regulatory protein Mena in direct contact with a perivascular, proangiogenic macrophage which expresses high levels of TIE2 and VEGF, where both of these cells are tightly bound to a blood vessel endothelial cell. Cancer cells can intravasate through TMEM doorways due to transient vascular permeability orchestrated by the joint activity of the TMEM-associated macrophage and the TMEM-associated Mena-expressing cancer cell. In this manuscript, we describe two methods for assessment of TMEM-mediated transient vascular permeability: intravital imaging and fixed tissue immunofluorescence. Although both methods have their advantages and disadvantages, combining the two may provide the most complete analyses of TMEM-mediated vascular permeability as well as microenvironmental prerequisites for TMEM function. Since the metastatic process in breast cancer, and possibly other types of cancer, involves cancer cell dissemination via TMEM doorways, it is essential to employ well established methods for the analysis of the TMEM doorway activity. The two methods described here provide a comprehensive approach to the analysis of TMEM doorway activity, either in na&#239;ve or pharmacologically treated animals, which is of paramount importance for pre-clinical trials of agents that prevent cancer cell dissemination via TMEM.

We describe two methods for assessing transient vascular permeability associated with tumor microenvironment of metastasis (TMEM) doorway function and cancer cell intravasation using intravenous injection of high-molecular weight (155 kDa) dextran in mice. The methods include intravital imaging in live animals and fixed tissue analysis using immunofluorescence.

利用生命内成像和固定组织分析评估与癌细胞传播相关的转移门道介导血管渗透性的肿瘤微环境

The most common cause of cancer related mortality is metastasis, a process that requires dissemination of cancer cells from the primary tumor to secondary ...

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 ...

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

Optical imaging offers a wide range of diagnostic modalities and has attracted a lot of interest as a tool for biomedical imaging. Despite the enormous number of imaging techniques currently available and the progress in instrumentation, there is still a need for highly sensitive probes that are suitable for in vivo imaging. One typical problem of available preclinical fluorescent probes is their rapid clearance in vivo, which reduces their imaging sensitivity. To circumvent rapid clearance, increase number of dye molecules at the target site, and thereby reduce background autofluorescence, encapsulation of the near-infrared fluorescent dye, DY-676-COOH in liposomes and verification of its potential for in vivo imaging of inflammation was done. DY-676 is known for its ability to self-quench at high concentrations. We first determined the concentration suitable for self-quenching, and then encapsulated this quenching concentration into the aqueous interior of PEGylated liposomes. To substantiate the quenching and activation potential of the liposomes we use a harsh freezing method which leads to damage of liposomal membranes without affecting the encapsulated dye. The liposomes characterized by a high level of fluorescence quenching were termed Lip-Q. We show by experiments with different cell lines that uptake of Lip-Q is predominantly by phagocytosis which in turn enabled the characterization of its potential as a tool for in vivo imaging of inflammation in mice models. Furthermore, we use a zymosan-induced edema model in mice to substantiate the potential of Lip-Q in optical imaging of inflammation in vivo. Considering possible uptake due to inflammation-induced enhanced permeability and retention (EPR) effect, an always-on liposome formulation with low, non-quenched concentration of DY-676-COOH (termed Lip-dQ) and the free DY-676-COOH were compared with Lip-Q in animal trials.

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

The use of fluorophores for in vivo imaging can be greatly limited by opsonization, rapid clearance, low detection sensitivity and cytotoxic effects on the host. Encapsulation of fluorophores in liposomes by film hydration and extrusion leads to fluorescence quenching and protection which enables in vivo imaging with high detection sensitivity.

荧光淬一个脂质体包封的近红外荧光团的作为一个工具<em&gt;在体内</em&gt;光学成像

Optical imaging offers a wide range of diagnostic modalities and has attracted a lot of interest as a tool for biomedical imaging. Despite the enormous ...

A Permanent Window for Investigating Cancer Metastasis to the Lung

Metastasis, accounting for ~90% of cancer-related mortality, involves the systemic spread of cancer cells from primary tumors to secondary sites such as the bone, brain, and lung. Although extensively studied, the mechanistic details of this process remain poorly understood. While common imaging modalities, including computed tomography (CT), positron emission tomography (PET), and magnetic resonance imaging (MRI), offer varying degrees of gross visualization, each lacks the temporal and spatial resolution necessary to detect the dynamics of individual tumor cells. To address this, numerous techniques have been described for intravital imaging of common metastatic sites. Of these sites, the lung has proven especially challenging to access for intravital imaging owing to its delicacy and critical role in sustaining life. Although several approaches have previously been described for single-cell intravital imaging of the intact lung, all involve highly invasive and terminal procedures, limiting the maximum possible imaging duration to 6-12 h. Described here is an improved technique for the permanent implantation of a minimally invasive thoracic optical Window for High-Resolution Imaging of the Lung (WHRIL). Combined with an adapted approach to microcartography, the innovative optical window facilitates serial intravital imaging of the intact lung at single-cell resolution across multiple imaging sessions and spanning multiple weeks. Given the unprecedented duration of time over which imaging data can be gathered, the WHRIL can facilitate the accelerated discovery of the dynamic mechanisms underlying metastatic progression and numerous additional biologic processes within the lung.

Here, we present a protocol for the surgical implantation of a permanently indwelling optical window for the murine thorax, which enables high-resolution, intravital imaging of the lung. The permanence of the window makes it well-suited to the study of dynamic cellular processes in the lung, especially those that are slowly evolving, such as metastatic progression of disseminated tumor cells.

研究癌症转移至肺部的永久窗口

Metastasis, accounting for ~90% of cancer-related mortality, involves the systemic spread of cancer cells from primary tumors to secondary sites such as ...

制备基于微流体的血管化人类微肿瘤模型

Establishing a Physiologic Human Vascularized Micro-Tumor Model for Cancer Research

A lack of validated cancer models that recapitulate the tumor microenvironment of solid cancers in vitro remains a significant bottleneck for preclinical cancer research and therapeutic development.&#160;To overcome this problem, we have developed the vascularized microtumor (VMT), or tumor chip, a microphysiological system that realistically models the complex human tumor microenvironment. The VMT forms de novo within a microfluidic platform by co-culture of multiple human cell types under dynamic, physiological flow conditions. This tissue-engineered micro-tumor construct incorporates a living perfused vascular network that supports the growing tumor mass just as newly formed vessels do in vivo. Importantly, drugs and immune cells must cross the endothelial layer to reach the tumor, modeling in vivo physiological barriers to therapeutic delivery and efficacy. Since the VMT platform is optically transparent, high-resolution imaging of dynamic processes such as immune cell extravasation and metastasis can be achieved with direct visualization of fluorescently labeled cells within the tissue. Further, the VMT retains in vivo tumor heterogeneity, gene expression signatures, and drug responses. Virtually any tumor type can be adapted to the platform, and primary cells from fresh surgical tissues grow and respond to drug treatment in the VMT, paving the way toward truly personalized medicine. Here, the methods for establishing the VMT and utilizing it for oncology research are outlined. This innovative approach opens new possibilities for studying tumors and drug responses, providing researchers with a powerful tool to advance cancer research.

作者聚焦：创建人类血管化微肿瘤作为转化癌症研究的模型 

This protocol presents a physiologically relevant tumor-on-a-chip model to perform high-throughput basic and translational human cancer research, advancing drug screening, disease modeling, and personalized medicine approaches with a description of loading, maintenance, and evaluation procedures.

建立用于癌症研究的生理性人血管化微肿瘤模型

A lack of validated cancer models that recapitulate the tumor microenvironment of solid cancers in vitro remains a significant bottleneck for preclinical ...

小鼠的引导肿瘤接种和多模式成像

A Dorsal Skinfold Window Chamber Tumor Mouse Model for Combined Intravital Microscopy and Magnetic Resonance Imaging in Translational Cancer Research

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the tumor microenvironment and its response to therapy. These imaging modalities have micron-scale resolution but have limited use in the clinic due to their shallow penetration depth into tissue. More clinically applicable imaging modalities such as CT, MRI, and PET have much greater penetration depth but have comparatively lower spatial resolution (mm scale). 
To translate preclinical intravital imaging findings into the clinic, new methods must be developed to bridge this micro-to-macro resolution gap. Here we describe a dorsal skinfold window chamber tumor mouse model designed to enable preclinical intravital and clinically applicable (CT and MR) imaging in the same animal, and the image analysis platform that links these two disparate visualization methods. Importantly, the described window chamber approach enables the different imaging modalities to be co-registered in 3D using fiducial markers on the window chamber for direct spatial concordance. This model can be used for validation of existing clinical imaging methods, as well as for the development of new ones through direct correlation with "ground truth" high-resolution intravital findings. 
Finally, the tumor response to various treatments-chemotherapy, radiotherapy, photodynamic therapy-can be monitored longitudinally with this methodology using preclinical and clinically applicable imaging modalities. The dorsal skinfold window chamber tumor mouse model and imaging platforms described here can thus be used in a variety of cancer research studies, for example, in translating preclinical intravital microscopy findings to more clinically applicable imaging modalities such as CT or MRI.

作者聚焦:整合高分辨率活体成像和 MRI 以增强立体定向放射治疗计划

Translation of Intravital microscopy findings is challenged by its shallow depth penetration into tissue. Here we describe a dorsal window chamber mouse model that enables co-registration of intravital microscopy and clinically applicable imaging modalities (e.g., CT, MRI) for direct spatial correlation, potentially streamlining clinical translation of intravital microscopy findings.

背侧皮褶窗室肿瘤小鼠模型在转化癌症研究中用于联合活体显微镜和磁共振成像

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the ...

灌注的空间测量，实体肿瘤间质流体压力和脂质体积累

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超声造影有针对性的治疗胶质瘤的小鼠通过药物轴承纳米粒子交付和微血管消融

使用校准动脉自旋标记磁共振肺灌注显像定量

利用生命内成像和固定组织分析评估与癌细胞传播相关的转移门道介导血管渗透性的肿瘤微环境

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

荧光淬一个脂质体包封的近红外荧光团的作为一个工具

研究癌症转移至肺部的永久窗口

建立用于癌症研究的生理性人血管化微肿瘤模型

建立用于癌症研究的生理性人血管化微肿瘤模型

建立用于癌症研究的生理性人血管化微肿瘤模型

背侧皮褶窗室肿瘤小鼠模型在转化癌症研究中用于联合活体显微镜和磁共振成像