这项工作已经由国家癌症研究所的资助以R01 CA120368，以R01 CA109558（国民党，缩减工时，护送队，BWP），RO1的CA132750（兆焦耳，BWP）和K25中CA138578（佛罗里达州）的资助，加拿大健康研究的博士后奖学金获奖研究院（国民党）。部分资金由高级研究技术（蒙特利尔，QC）的荧光断层成像系统的发展。

1。动物的制备<ol><li>麻醉裸鼠腹腔内注射氯胺酮 - 甲苯噻嗪（100毫克/千克10毫克/公斤的IP）（查尔斯河，威尔明顿，马萨诸塞州）。 </li><li>鼠标放置在立体框架，使一个切口，将头皮上的头骨左侧，使用18号针头，创建头骨2毫米到1毫米直径的孔从中央线和2毫米的背后前囟门。 </li><li> 5μL磷酸盐缓冲溶液注入5×10 5的U251人类神经胶质瘤细胞（请在达特茅斯学院，汉诺威，NH博士马克以色列提供）到左侧大脑半球表面以下约2毫米的深度大脑。使用一个Hamilton微量注射器18钝截至细胞植入27号针头插入针从头骨的外表面3毫米的一角，然后退出1毫米到创建一个细胞的口袋。 </li><li>缝合切口部位，并允许重新covery从手术。 </li><li>等待〜14天，使肿瘤生长前成像。 </li></ol> 2。荧光体层摄影术系统标定<ol><li>鼠标成像的日子，启动系统和激光器和光探测器，允许约20分钟热身，以避免在系统的灵敏度漂移。 </li><li>放置一个100°×4°在直接成像龙门中心工程线扩散器（Thorlabs，牛顿，新泽西州），正常的激励激光皮秒脉冲80MHz的多模635 nm激光二极管（PicoQuant光子北美国公司，西田，马）。扩散器的角度调整，以最大限度地发挥所有五个光收集渠道检测到的信号量。提供一个完整的描述成像几何别处11,14,15。 </li><li>将外径2中性密度过滤器在前面的所有荧光检测的光电倍增管（PMT）和外径1中性密度过滤器（Thorlabs，牛顿，新泽西州）（THorlabs，牛顿，新泽西州），在前面所有透过率检测的光电倍增管。收集100时空传播的激光脉冲型材（TPSF），每个1的整合时间。 </li><li>正常化的激光参考每个TPSF，正确的时空漂移在激光参考，超过平均每个探测器的所有迭代。这些的平均TPSFs是探测器特定的仪器响应函数（IRF）的光学图像重建中使用。 </li></ol> 3。成像议定书" <ol><li>麻醉与异氟醚2％的氧气（1升/分钟）的鼠标。 </li><li>注入100μL磷酸盐缓冲液，腹腔，12小时前1 nanomole的IRDye 800CW-EGF的Alexa Fluor 647和1 nanomole针对表皮生长因子受体的过度表达，在肿瘤成像。 </li><li>鼠标放置到玻璃纤维成像床支持，安排的鼠标，所以它的鼻子仍然在提供异氟醚麻醉锥。 </li><li>恩确保鼠标定位: 即在床上适当的荧光断层成像系统固定在床上时鼠标是在的成像龙门近似中心。有关鼠标的激发激光180°旋转，确保激光的焦点照亮中心鼠标一点，大约从激光在各个角度的角度来看，这种定位可以指导。 </li><li>一旦定位，仔细转移成像床和鼠标的微型电脑（探索轨迹，GE医疗，伦敦，ON）扫描仪，并收集在鼠标的整个头的分辨率为93微米的各向同性的解剖信息。 </li><li>可视化CT图像堆栈和选择（S）与荧光断层成像系统成像切片。 </li><li>仔细转让荧光断层成像系统成像床和鼠标。选择源位置的编号，以收集有关鼠标的数据，为每个成像SLICE（32），积分时间为每个TPSF测量（1秒），为每个源的位置（10）的迭代的数量，并从步骤3.6 CT图像堆栈中的地位和所需的影像片的数量。括号中的数字是每个成像产生每成像片〜5分钟的数据采集参数的典型值。 </li><li>放置在前面的荧光检测的光电倍增管的三重缺口过滤器（色度科技公司，波纹管瀑布，VT），以限制任何达到荧光探测器的激光，外径2中性密度滤光片的透过率检测的光电倍增管前，以避免饱和这些探测器。 </li><li>运行数据采集软件，收集在每个定义的源，探测器位置和荧光激发波长（635 nm和755 nm的激发的Alexa Fluor 647和IRDye 800CW-EGF的示踪剂，分别）为每个和透光TPSFs的。为每一套TPSFs收集，监视和记录的激光强度参考的PMT通道。 </li></ol> 4。图像重建<ol><li>确定鼠标的外表面和成像床支撑杆从CT图像的位置，并创建口罩，分别涵盖了鼠标的束缚，成像棒。 </li><li>使用鼠标面具产生了使用的NIRFAST软件19动物的有限元网格。 </li><li>本地化基于微型电脑和荧光的空间登记统筹20网格表面上的荧光断层成像系统的源和探测器的位置。 </li><li>卸下光学数据源或探测器的位置成像床支撑杆的位置交互点。 </li><li>规范化数据收集在每个源探测器的位置由激光参考，正确的时间漂移​​在激光参考，正确的过滤器的灵敏度，这是由实验测试确定<html>在购买时间15 </li><li>每个源探测器位置的数据的出生比率（荧光分为透光）和繁殖与动物统一的光学性质的有限元网格上的透光率的正向模型的仿真。这样做，以减轻相关的错误源或探测器组织耦合21，校准数据模型22，和数据调整等方面的数据模型不匹配23,24。 </li><li>构建一个数据向量组成的出生比例收集到的数据在两个波长的规模差异。比例因子的选择，以最大限度地提高EGFR结合对比。执行时域与校准的区别使用TPSF的数据作为输入的每个检测通道的图像重建，并建立荧光增强针对性示踪15地图。 </li></ol> 5。代表结果"&gt;荧光重建（ 图1b确定脑胶质瘤的质量中心共同注册的鼠标在图1b 1 U251细胞原位脑胶质瘤肿瘤与头部CT的解剖图像叠加荧光重建的一个例子。 ）CT和MRI图像质量对比增强磁共振成像（ 图1a）确定的肿瘤中心内1毫米。共登记基于一个共同的信息化改造。 <img src="/files/ftp_upload/4050/4050fig1.jpg" alt="图1" /> 图1。对比度增强（钆）鼠标头部的磁共振成像（一）。鼠标用的U251人脑胶质瘤细胞株接种原位。吸收超过正常脑造影剂，肿瘤的位置，可以看到在左侧大脑半球（右图像）和白色箭头指示。 "都有对应Ğ计算机断层扫描图像（从同一位置上的鼠标头），（二）与表皮生长因子有针对性的荧光减去不相关的荧光重建覆盖描绘。荧光的单位是在逆毫米和涉及到的结合针对性的荧光量子效率和乘以其浓度的吸收系数。

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没有利益冲突的声明。

荧光断层扫描（FT）是一个敏感，电离辐射自由分子成像方式的基础上通过生物组织的可见光和近红外光传输。在FT的利益大部分都集中在小动物实验模型1和重要的研究领域之一，一直是癌症的生物标志物的表达和26分子疗法的研究，以加速药物发现和开发的潜力。目前，有两个相互竞争的办法FT系统设计。最常见的设计是基于冷却的电荷耦合器件（CCD）摄像机荧光检测4-9。这种设计提供了一个高密度的测量，最大限度地组织取样，因为每个像素的CCD相机可以检测光线已走遍组织的一个独特的路径。然而，CCD相机有一个有限的动态范围和读出噪声限制了他们的最终灵敏度。第二个设计，避免了潜在的限制<html> CCD相机检测的tions采用高度敏感的单光子计数技术的基础上，这种探测器使用光电倍增管或雪崩光电二极管10-13的影响。这些更敏感的检测方法的缺点是，每个探测器可以只收集在一个单一的点光，因此，要实现组织密度采样，要么许多探测器，必须使用（这是非常昂贵），或许多预测成像具有相同探测器（可以是耗时）。小动物的FT组织取样的最佳水平，而没有商定，对案件逐案的基础上可能会有所不同，它同意，单光子计数仪器更适合探索金融时报"的敏感性限制条款它能够探测到低浓度的分子标记。在这项研究中，我们提供了一种方法，为开展金融时报"使用单光子计数检测仪器定位在小鼠的肿瘤。 耳鼻喉科"&gt;有涉及到生产与时间相关单光子计数金融时报"的强大的数据集的四个关键步骤。首先是一个合适的和简单的校准过程中的应用。在提出的方法，每个检测通道各自的敏感性占收集计量基准的激发光传输线扩散器通过旨在引导光等于每个探测器的分数15。此外，在实验中检测光不断校准激光参考，双方强度和平均时间，这可能会随着时间的推移而波动11,15激光参考通道的运作，第二个关键步骤是"金融时报数据的准确收集和解剖成像制导荧光重建共登记单没有提供解剖信息;因此，为了创造一个光传输模型可以用来重建的劳在试样表面荧光检测的标本内的荧光来源的阳离子，解剖标本FT系统必须准确知道。在我们的系统中，被收购的解剖信息与空间坐标空间已注册的FT系统15,20微型计算机断层扫描系统。第三个关键的一步是确保最佳曝光（ 即总光子探测时间为每个激光投影）受聘在每个源探测器的位置。这是很重要的原因有两个:第一，要确保有足够的信号与噪声在每个检测位置和第二，以避免探测器饱和，这可能会损坏检测单位。为了实现在每个探测器位置的最佳曝光，自动曝光控制，基本上从二，低信号暴露14三角测量的最佳曝光。第四个关键该方法的步骤是引用传输激发光量的荧光数据收集。此参照通常被称为出生比率，并提供金融时报"的许多好处，主要是减缓的数据模型不匹配的错误23,24之一。该系统的设计，同时检测窜，在每个检测通道分为2个独立的光电倍增管的光激发光荧光和传播。通过这样做，我们避免出生比率的准确性议案的任何影响。 它一方面与一个强大的数据集时域数据，图像重建涉及逆问题求解的有限元网格表达: D = JX 其中d是一个向量为N×M为N源，探测器的预测和 m TPSF时间门元素; J是一个 N x M--升灵敏度矩阵（或雅可比），L的网格节点; x是在每个节点上的荧光光学性质的载体，具有尺寸 L d是校准实验和 J期间收集的数据是模拟采用有限元解决方案。到时域扩散近似的荧光运输25。时间维的 J也探测器特定的仪器响应函数的卷积。X荧光地图利益的代表，并用Tikhonov正则化15 1的Levenberg-Marqardt非负最小二乘法解决。 这里的方法提出，它描述了一个程序化荧光标记的使用高度敏感的光子计数荧光检测小鼠肿瘤的能力，有潜力，推动金融时报的限制。在先前的研究中，采用这种潜在在较大的小鼠动物模型，如大鼠，以及在现有鼠标中小型标本系统设计，提高了灵敏度，方法，结果表明17。这种方法的直接应用，将小动物肿瘤模型体内一种高通量的手段，以评估药物疗效监测的生物标志物的表达。该系统在多个波长的荧光激发和检测的能力允许多个荧光标记物的同时检测。额外的荧光标记物提供了一种手段，审问的病理多个方面，同时，或可用于在这项研究中，双记者的方法测量在体内结合的潜力，受体密度的标记，如聘请更多的定量成像方法26,27。

<table border="1"><tbody><tr><td>试剂名称</td><td>公司</td><td>目录编号</td><td>评论（可选） </td></tr><tr><td> IRDye 800CW - 表皮生长因子</td><td>李-COR公司Biosciences公司</td><td> 926-08446 </td><td></td></tr><tr><td>的Alexa Fluor 647，琥珀酰亚胺酯</td><td>生命科技</td><td> A20106 </td><td>与水反应，以尽量减少非特异性结合</td></tr></tbody></table>

computed tomography-guided time-domain diffuse fluorescence tomography in small animals for localization of cancer biomarkers

小动物荧光分子成像（FMI）可以是一个强大的工具，临床前药物发现和开发研究1。然而，由组织生色团（如血红蛋白，水，脂肪，黑色素）的光吸收通常限制光信号的传播，通过厚度大于2几毫米。相比其他可见光波长，组织吸收红光和近红外（近红外）的光吸收显着降低和非弹性散射占主导地位的光组织的互动机制。荧光剂的吸收和发射光在近红外范围（600-1000 nm）的，相对近期的发展带动了成像系统和光的传播模式，可以实现在3小动物全身三维成像的发展。 尽管在这一领域的巨大进步，弥漫性荧光体层摄影术的病态性质仍然是一个重要的的稳定，相反​​复苏和空间分辨率图像重建技术，在FMI的小动物的最佳途径的问题还有待商定。大多数的研究小组已投资在电荷耦合器件（CCD）为基础的系统，提供丰富的组织取样，但最理想的敏感性4-9，而我们组和10-13少数人的追求非常高的灵敏度探测器的基础上的系统，允许组织密度采样，此时要实现成本低成像吞吐量。在这里，我们证明申请单光子荧光断层成像系统的检测技术，本地化癌细胞在小鼠模型脑损伤的方法。 荧光断层成像（FT）系统采用单光子计数用光电倍增管（PMT）和信息丰富的时域光检测在非接触式构象11。这提供了一个同步的山坳激发和发射光传输的经文，包括激发自动曝光控制14，激光参照，与小动物计算机断层扫描系统（微型电脑）15，共登记。裸鼠模型被用于成像。原位接种动物与人脑胶质瘤细胞株（U251细胞）在左侧大脑半球，2周后成像。肿瘤注射荧光示踪剂荧光，IRDye 800CW表皮生长因子（Li-COR Biosciences公司，林肯，NE）的有针对性的表皮生长因子受体，细胞膜蛋白已知U251肿瘤行过度和许多其他癌症18。第二，没有针对性的荧光示踪的Alexa Fluor 647（Life Technologies公司，大岛，NY）也被注入帐户针对性示踪剂的摄取提供量化示踪结合和受体的可用性/密度的一种手段，非受体介导的效果27。一个CT引导下，TIME域算法，用于重建的两个荧光示踪剂的位置（ 即 ，肿瘤的位置）在小鼠的大脑和他们的能力，本地化的肿瘤对比增强磁共振成像验证。 虽然荧光成像在脑胶质瘤的小鼠模型表明，在这部影片中提出的方法可以扩展到不同的肿瘤模型中的各种小动物模型可能上升到老鼠的大小17。

Watch this Scientific Journal Video about Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers at JoVE.com

Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

弥漫性荧光体层摄影术提供了一个相对较低的成本和潜在的高通量的方法，以临床<em&gt;在体内</em&gt;肿瘤显像。计算机断层扫描引导非接触式时域使用荧光针对肿瘤标志物的表皮生长因子受体在小鼠脑胶质瘤模型系统的光学数据收集，校准和图像重建方法。

电脑断层导引的时域漫荧光体层摄影术在小动物的肿瘤生物标志物的本土化

Small animal fluorescence molecular imaging (FMI) can be a powerful tool for preclinical drug discovery and development studies1. However, light ...

computed-tomography-guided-time-domain-diffuse-fluorescence

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.3K 次观看.  Dartmouth College. 弥漫性荧光体层摄影术提供了一个相对较低的成本和潜在的高通量的方法，以临床在体内肿瘤显像。计算机断层扫描引导非接触式时域使用荧光针对肿瘤标志物的表皮生长因子受体在小鼠脑胶质瘤模型系统的光学数据收集，校准和图像重建方法。

视频: Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location.
Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.

An experimental mouse model of bone metastasis was established following intracardiac delivery of luciferase expressing mammary tumor cells. Tumor development and resulted osteolytic lesion were monitored longitudinally with bioluminescence and micro CT imaging.

生物发光和微型CT成像监测肿瘤转移和溶骨性病变

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

科研

医学

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

Metastasis is responsible for most cancer deaths. Despite extensive research, the mechanistic understanding of the complex processes governing metastasis remains incomplete. In vivo models are paramount for metastasis research, but require refinement. Tracking spontaneous metastasis by non-invasive in vivo imaging is now possible, but remains challenging as it requires long-time observation and high sensitivity. We describe a longitudinal combined radionuclide and fluorescence whole-body in vivo imaging approach for tracking tumor progression and spontaneous metastasis. This reporter gene methodology employs the sodium iodide symporter (NIS) fused to a fluorescent protein (FP). Cancer cells are engineered to stably express NIS-FP followed by selection based on fluorescence-activated cell sorting. Corresponding tumor models are established in mice. NIS-FP expressing cancer cells are tracked non-invasively in vivo at the whole-body level by positron emission tomography (PET) using the NIS radiotracer [18F]BF4-. PET is currently the most sensitive in vivo imaging technology available at this scale and enables reliable and absolute quantification. Current methods either rely on large cohorts of animals that are euthanized for metastasis assessment at varying time points, or rely on barely quantifiable 2D imaging. The advantages of the described method are: (i) highly sensitive non-invasive in vivo 3D PET imaging and quantification, (ii) automated PET tracer production, (iii) a significant reduction in required animal numbers due to repeat imaging options, (iv) the acquisition of paired data from subsequent imaging sessions providing better statistical data, and (v) the intrinsic option for ex vivo confirmation of cancer cells in tissues by fluorescence microscopy or cytometry. In this protocol, we describe all steps required for routine NIS-FP-afforded non-invasive in vivo cancer cell tracking using PET/CT and ex vivo confirmation of in vivo results. This protocol has applications beyond cancer research whenever in vivo localization, expansion and long-time monitoring of a cell population is of interest.

We describe a protocol for preclinical in vivo tracking of cancer metastasis. It is based on a radionuclide-fluorescence reporter combining the sodium iodide symporter, detected by non-invasive [18F]tetrafluoroborate-PET, and a fluorescent protein for streamlined ex vivo confirmation. The method is applicable for preclinical in vivo cell tracking beyond tumor biology.

核素荧光报告基因成像在啮齿动物肿瘤模型中跟踪肿瘤进展

Metastasis is responsible for most cancer deaths. Despite extensive research, the mechanistic understanding of the complex processes governing metastasis ...

癌症研究

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting immunodeficient mice subcutaneously with cancer cells and measuring the tumor mass after it becomes visible and palpable. Orthotopic injections of cancer cells aim to introduce the xenograft in the microenvironment that most closely resembles the tissue of origin of the tumor being studied. Brain cancer research requires intracranial injection of cancer cells to allow the tumor formation and analysis in the unique microenvironment of the brain. The in vivo imaging of intracranial xenografts monitors instantaneously the tumor mass of orthotopically engrafted mice. Here we report the use of fluorescence molecular tomography (FMT) of brain tumor xenografts. The cancer cells are first transduced with near infrared fluorescent proteins and then injected in the brain of immunocompromised mice. The animals are then scanned to obtain quantitative information about the tumor mass over an extended period of time. Cell pre-labeling allows for cost effective, reproducible, and reliable quantification of the tumor burden within each mouse. We eliminated the need for injecting imaging substrates, and thus reduced the stress on the animals. A limitation of this approach is represented by the inability to detect very small masses; however, it has better resolution for larger masses than other techniques. It can be applied to evaluate the efficacy of a drug treatment or genetic alterations of glioma cell lines and patient-derived samples.

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Orthotopic intracranial injection of tumor cells has been used in cancer research to study brain tumor biology, progression, evolution, and therapeutic response. Here we present fluorescence molecular tomography of tumor xenografts, which provides real-time intravital imaging and quantification of a tumor mass in preclinical glioblastoma models.

胶质瘤移植中体内成像的荧光分子层析成像研究

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting ...

Multimodal Bioluminescent and Positronic-emission Tomography/Computational Tomography Imaging of Multiple Myeloma Bone Marrow Xenografts in NOG Mice

Multiple myeloma (MM) tumors engraft in the bone marrow (BM) and their survival and progression are dependent upon complex molecular and cellular interactions that exist within this microenvironment. Yet the BM microenvironment cannot be easily replicated in vitro, which potentially limits the physiologic relevance of many in vitro and ex vivo experimental models. These issues can be overcome by utilizing a xenograft model in which luciferase (LUC)-transfected 8226 MM cells will specifically engraft in the mouse skeleton. When these mice are given the appropriate substrate, D-luciferin, the effects of therapy on tumor growth and survival can be analyzed by measuring changes in the bioluminescent images (BLI) produced by the tumors in vivo. This BLI data combined with positronic-emission tomography/computational tomography (PET/CT) analysis using the metabolic marker 2-deoxy-2-(18F)fluoro-D-glucose (18F-FDG) is used to monitor changes in tumor metabolism over time. These imaging platforms allow for multiple noninvasive measurements within the tumor/BM microenvironment.

Here we use bioluminescent, X-ray, and positron-emission tomography/computed tomography imaging to study how inhibiting mTOR activity impacts bone marrow-engrafted myeloma tumors in a xenograft model. This allows for physiologically relevant, non-invasive, and multimodal analyses of the anti-myeloma effect of therapies targeting bone marrow-engrafted myeloma tumors in vivo.

nog 小鼠多发性骨髓瘤异种移植的多模态生物发光和正电子发射成形断层扫描成像

Multiple myeloma (MM) tumors engraft in the bone marrow (BM) and their survival and progression are dependent upon complex molecular and cellular ...

Hybrid &#181;CT-FMT imaging and image analysis

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to assess the biodistribution of novel probes and to assess disease progression using established molecular probes or reporter genes. The combination with an anatomical modality, e.g., micro computed tomography (&#181;CT), is beneficial for image analysis and for fluorescence reconstruction. We describe a protocol for multimodal &#181;CT-FMT imaging including the image processing steps necessary to extract quantitative measurements. After preparing the mice and performing the imaging, the multimodal data sets are registered. Subsequently, an improved fluorescence reconstruction is performed, which takes into account the shape of the mouse. For quantitative analysis, organ segmentations are generated based on the anatomical data using our interactive segmentation tool. Finally, the biodistribution curves are generated using a batch-processing feature. We show the applicability of the method by assessing the biodistribution of a well-known probe that binds to bones and joints.

Hybrid  &#181;CT-FMT imaging and image analysis

We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (&#181;CT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.

混合μCT-FMT成像和图像分析

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to ...

生物工程

Using Micro-computed Tomography for the Assessment of Tumor Development and Follow-up of Response to Treatment in a Mouse Model of Lung Cancer

Lung cancer is the most lethal cancer in the world. Intensive research is ongoing worldwide to identify new therapies for lung cancer. Several mouse models of lung cancer are being used to study the mechanism of cancer development and to experiment with various therapeutic strategies. However, the absence of a real-time technique to identify the development of tumor nodules in mice lungs and to monitor the changes in their size in response to various experimental and therapeutic interventions hampers the ability to obtain an accurate description of the course of the disease and its timely response to treatments. In this study, a method using a micro-computed tomography (CT) scanner for the detection of the development of lung tumors in a mouse model of lung adenocarcinoma is described. Next, we show that monthly follow-up with micro-CT can identify dynamic changes in the lung tumor, such as the appearance of additional nodules, increase in the size of previously detected nodules, and decrease in the size or complete resolution of nodules in response to treatment. Finally, the accuracy of this real-time assessment method was confirmed with end-point histological quantification. This technique paves the way for planning and conducting more complex experiments on lung cancer animal models, and it enables us to better understand the mechanisms of carcinogenesis and the effects of different treatment modalities while saving time and resources.

We describe a method for the detection of tumor nodule development in the lungs of an adenocarcinoma mouse model using micro-computed tomography and its use for monitoring changes in nodule size over time and in response to treatment. The accuracy of the assessment was confirmed with end-point histological quantification.

采用微计算机断层扫描对肿瘤发展的评估和后续反应来治疗肺癌的小鼠模型

Lung cancer is the most lethal cancer in the world. Intensive research is ongoing worldwide to identify new therapies for lung cancer. Several mouse ...

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not routinely employed in animal models. Conventional experimental readouts often rely on post mortem and ex vivo analyses, which prevent intra-individual follow-up examinations and increase the number of study animals needed. Fluorescence-mediated tomography enables the non-invasive, repetitive, quantitative, three-dimensional assessment of fluorescent probes. It is highly sensitive and permits the use of molecular makers, which allows for the specific detection and characterization of distinct molecular targets. In particular, targeted probes represent an innovative tool for analyzing gene activation and protein expression in inflammation, autoimmune disease, infection, vascular disease, cell migration, tumorigenesis, etc. In this article, we provide step-by-step instructions on this sophisticated imaging technology for the in vivo detection and characterization of inflammation (i.e., F4/80-positive macrophage infiltration) in a widely used murine model of intestinal inflammation. This technique might also be used in other research areas, such as immune cell or stem cell tracking.

Target-specific probes represent an innovative tool for analyzing molecular mechanisms, such as protein expression in various types of disease (e.g., inflammation, infection, and tumorigenesis). In this study, we describe a quantitative three-dimensional tomographic assessment of intestinal macrophage infiltration in a murine model of colitis using F4/80-specific fluorescence-mediated tomography.

荧光介导的断层扫描对巨噬细胞相关小鼠肠道炎症的检测和定量

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

In Vivo Fluorescence Imaging to Localize Antibodies in a Mouse Tumor Xenograft Model

Source: Shivange, G. et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/60727">Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody.</a>&#160;J. Vis. Exp. (2020)
The video demonstrates a fluorescence imaging method for in vivo antibody localization in a mouse tumor xenograft model. It involves the injection of tumor cells in a mouse to generate solid xenografts beneath the skin. Subsequent injection of fluorescent antibodies confirms specific localization through enhanced fluorescence signals within the tumor xenograft.

体内荧光成像定位小鼠肿瘤异种移植模型中的抗体

1. Expression and purification of antibodies

	Maintenance of CHO cells
	
		Grow CHO cells in FreeStyle CHO Media supplemented with commercially available ...

JoVE Encyclopedia of Experiments

Immunology

Antibody-Based Technologies

Antibody-Based Imaging and Detection Methods

FDG Based PET/CT Scan: A Method of Using Radiotracer for Non-invasive Imaging of Tumor Metabolism

Source: Momcilovic M. et al., <a href="https://jove.unicartagenaproxy.elogim.com/v/57167/utilizing-18f-fdg-petct-imaging-quantitative-histology-to-measure">Utilizing&#160;18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer.</a> J. Vis. Exp. (2018)
This article describes multimodality imaging of cancerous tissues with PET and CT diagnostic imaging by using a radiotracer 18F-FDG. Cancerous tissue with high glycolytic activity uptakes more 18F-FDG.

基于 FDG 的 PET/CT 扫描:一种使用放射性示踪剂进行肿瘤代谢无创成像的方法

Source: Momcilovic M. et al., Utilizing&#160;18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in ...

肺癌

In vivo study

Fluorescence Molecular Tomography: An Imaging Technique for In Vivo Imaging of Fluorescent Protein-tagged Glioblastoma Xenografts in Mouse Model

Source: Benitez, J. A. et al., <a href="https://jove.unicartagenaproxy.elogim.com/video/57448" target="_blank">Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts.</a> J. Vis. Exp. (2018)
In this video, we describe fluorescence molecular tomography or FMT for in vivo imaging of fluorescent protein-tagged glioblastoma xenografts in mouse models. The technique is useful for elucidating the effects of anti-tumor therapeutics at the molecular level.

荧光分子断层扫描:一种在小鼠模型中对荧光蛋白标记的胶质母细胞瘤异种移植物进行体内成像的成像技术

Source: Benitez, J. A. et al., Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts. J. Vis. Exp. (2018)
In this video, we ...