Name: in vivo immunofluorescence localization for assessment of therapeutic and diagnostic antibody biodistribution in cancer research
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我们感谢安德鲁·奥尔森博士（斯坦福神经科学显微镜服务）的讨论和设备使用。我们感谢尤尔根·威尔曼博士的导师。这项研究得到了NIH R21EB022214助学金（KEW）、NIH R25CA118681培训补助金（KEW）和NIH K99EB023279（KEW）的支持。斯坦福神经科学显微镜服务由NIH NS069375提供支持。

本文描述的所有方法均已获得斯坦福大学实验室动物护理机构行政小组（APLAC）的批准。
1. 乳腺癌发育的转基因小鼠模型
<ol><li>在继续之前，通过触觉或卡钳测量，从所需的癌症模型中观察小鼠，以进行适当的肿瘤生长。 注：这里使用了乳腺癌发育的转基因鼠模型（FVB/N-Tg（MMTV-PyMT）634Mul/J（MMTV-PyMT）。这些动物自发地在每个乳腺20个6至12周的年龄之间发...

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此方法有几个关键步骤，需要可能的修改以确保成功实施。首先，抗体/抗体结合静脉注射的剂量和时间必须针对具体应用进行定制。通常，应使用与抗体结合通常使用方式一致的剂量，即匹配治疗性抗体或基于抗体的造影剂的剂量。此外，应仔细考虑收集目标组织的时间。抗体和抗体结合剂比标准药物和造影剂具有更长的循环时间，但是，这些具有高度可变性，应针对所需应用进行优化，但建议...

单克隆抗体（mAb）是免疫球蛋白超级家族的大糖蛋白（约150 kDa），由B细胞分泌，在免疫系统中具有识别和抑制生物功能的主要功能，或标记破坏，细菌或病毒病原体，并能识别癌细胞上的异常蛋白质表达1。抗体可以有极高的亲和力，其特定表位下降到女性聚类浓度，使他们在生物医学2中非常有前途的工具。随着米尔斯坦和克勒（1984年获得诺贝尔奖）的混合瘤技术的发展，mAbs的生产成为可能。后来，人类mAbs产生使用噬菌体显示技术或转基因小鼠菌株，并彻底改变了它们作为新的研究工具和治疗4，5。
癌症是一个世界性的健康问题，也是主要死亡原因，因此需要新的预防、发现和治疗方法。迄今为止，mAbs已经允许免除基因及其蛋白质在肿瘤发生中的作用，当针对癌症生物标志物时，可以促进肿瘤的检测和患者分层的表征。对于癌症治疗，双特异性mAbs，抗体药物结合，和较小的抗体片段正在开发作为治疗，并针对药物交付，以提高疗效7。此外，抗体还用于分子成像模式（如荧光引导手术、光声 （PA） 成像、超声 （US） 分子成像和临床使用正电子发射）的造影剂的生物标记靶向断层扫描 （PET） 或单光子发射计算机断层扫描 （SPECT）8.最后，抗体也可用作治疗剂，使患者分层和反应监测的靶向疗法9。因此，新型mAbs开始在癌症的检测、诊断和治疗中发挥关键作用。
尽管在开发和生产新型和高度特异性的 mAbs 方面取得了重大进展，但由于肿瘤环境的复杂性，诊断和治疗应用可能会变得无效。抗体相互作用取决于表位的类型，即，无论是线性的还是构象的10。除了抗原的识别，抗体需要克服自然障碍，如血管壁，基底膜和肿瘤频闪，以达到表达抗原的目标细胞。抗体不仅通过可变片段抗原结合（Fab）域与组织相互作用，而且通过恒定的晶体片段（Fc）与组织相互作用，从而进一步导致场外相互作用11。靶向也由于肿瘤标记物在整个肿瘤体积的异质表达和肿瘤血管化和淋巴系统12、13的异质性而变得复杂。此外，肿瘤微环境由支持肿瘤细胞的癌症相关成纤维细胞、抑制抗肿瘤免疫反应的肿瘤免疫细胞以及支持氧气和营养物质输送的肿瘤内皮组成，干扰基于抗体的治疗或诊断的渗透、分布和可用性。总体而言，这些考虑可以限制治疗或诊断效果，降低治疗反应，并可能导致肿瘤耐药性。
因此，为了开发有效的基于抗体的疗法和诊断，评估基于抗体的偶联在肿瘤微环境中的生物分布和相互作用至关重要。目前，在临床前研究中，通过肿瘤14节的免疫荧光（IF）染色，对肿瘤研究模型中的标记表达进行了分析。标准 IF 染色使用主要标记特异性抗体执行，然后通过从动物分离的体外肿瘤组织切片上的二级荧光标记抗体突出显示这些抗体。此技术突出显示标记在组织固定时的静态位置，并且无法深入了解基于抗体的治疗或诊断如何在生理条件下分布或相互作用。PET、SPECT、US和PA的分子成像可以提供有关活前临床模型8、15中抗体结合造影剂分布的信息。由于这些成像模式是非侵入性的，可以进行纵向研究，并且每组动物数量最少，可以收集时间敏感数据。然而，这些非侵入性分子成像方法不够敏感，没有足够的分辨率来定位细胞层面的抗体分布。此外，由于造影剂16的偶联，原抗体的物理和生物特性可能发生巨大变化。
为了考虑体内生理和病理状况，考虑基于抗体的治疗和诊断如何在肿瘤环境中相互作用，并获得高分辨率细胞甚至亚细胞分布非结合抗体的配置文件，我们提出了一个IF方法，被认为是在Vivo免疫荧光定位（IVIL），其中抗原特异性抗体静脉注射在体内。基于抗体的治疗或诊断，作为主要抗体，在功能性血管中循环，并在高精度、活的肿瘤环境中与其目标蛋白结合。用原抗体分离出体内标记的肿瘤后，使用二次抗体来本地化累积和保留的抗体结合。这种方法类似于前面描述的IF组织学方法注射荧光标记抗体17。虽然在这里，使用非结合抗体可以避免由抗体修饰引起的生物分布特性的潜在变化。此外，荧光二级抗体的外生应用可避免组织收集和处理过程中荧光信号的可能丢失，并提供荧光信号强度的放大。我们的贴标方法反映了抗体药物和目标药物的体内生物分布，可为新型诊断和治疗剂的开发提供重要见解。
在这里，我们描述了IVIL方法在以往研究中应用的两种应用，这些研究研究抗体基造影剂的生物分布和可及性用于乳腺癌检测的分子成像方法。首先，抗体近红外染料结合（抗B7-H3抗体结合近红外荧光染料，青黄绿，B7-H3-ICG）和等型对照剂（一等型控制剂）用于荧光和光声分子成像被探索18。该协议中介绍了此应用程序的方法。其次，对netrin-1的构象敏感抗体的生物分布结果，通常不能用传统的IF成像检测，用于超声分子成像，在具有代表性的结果19中量化和呈现。在本文结束时，读者应该对采用IVIL方法进行基于抗体的研究应用感到自在。

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			<td height="21" style="height:21px;width:295px;">Animal Model</td>
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			<td height="21" style="height:22px;">FVB/N-Tg(MMTV-PyMT)634Mul/J</td>
			<td style="width:196px;">The Jackson Laboratory</td>
			<td style="width:166px;">002374</td>
			<td>Females, 4-6 weeks of age</td>
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			<td height="21" style="height:21px;width:295px;">Animal Handling Supplies</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">27G Catheter</td>
			<td style="width:196px;">VisualSonics</td>
			<td style="width:166px;">Please call to order</td>
			<td>Vevo MicroMarker Tail Vein Access Cannulation Kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Alcohol Wipes</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">22-246073</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Gauze Sponges (4&#34; x 4&#34; 16 Ply)</td>
			<td style="width:196px;">Cardinal Health</td>
			<td style="width:166px;">2913</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Heat Lamp</td>
			<td style="width:196px;">Morganville Scientific</td>
			<td style="width:166px;">&#160;HL0100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Isoflurane</td>
			<td style="width:196px;">Henry Schein Animal Health</td>
			<td style="width:166px;">29404</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Ophthalmic Ointment</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">NC0490117</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Surgical Tape</td>
			<td style="width:196px;">3M</td>
			<td style="width:166px;">1530-1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Tissue Collection</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Disposable Base Molds</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">22-363-556</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:295px;">Optimal Cutting Temperature (OCT) Medium</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">23-730-571</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:295px;">Surgical London Forceps</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td>11080-02</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:295px;">Surgical Scissors</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:166px;">14084-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Antibodies</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AlexaFluor-488 goat anti-rat IgG</td>
			<td style="width:196px;">Life Technologies</td>
			<td style="width:166px;">A-11006</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AlexaFluor-546 goat anti-rabbit IgG</td>
			<td style="width:196px;">Life Technologies</td>
			<td style="width:166px;">A-11010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AlexaFluor-594 goat anti-human IgG</td>
			<td style="width:196px;">Life Technologies</td>
			<td style="width:166px;">A11014</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Human IgG Isotype Control</td>
			<td style="width:196px;">Novus Biologicals</td>
			<td style="width:166px;">NBP1-97043</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Humanized anti-netrin-1 antibody&#160;</td>
			<td style="width:196px;">Netris Pharma</td>
			<td style="width:166px;"></td>
			<td>contact@netrispharma.com</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Rabbit anti-Mouse CD276 (B7-H3)</td>
			<td style="width:196px;">Abcam</td>
			<td style="width:166px;">ab134161</td>
			<td>EPNCIR122 Clone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Rat anti-Mouse CD31</td>
			<td style="width:196px;">BD Biosciences</td>
			<td style="width:166px;">550274</td>
			<td>MEC 13.3 Clone</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Reagents</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:196px;">Sigma-Aldrich</td>
			<td style="width:166px;">A2153-50G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Clear Nail Polish</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td>Any local drug store</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Indocyanine Green - NHS</td>
			<td style="width:196px;">Intrace Medical</td>
			<td style="width:166px;">ICG-NHS ester</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Mounting Medium</td>
			<td style="width:196px;">ThermoFisher Scientific</td>
			<td style="width:166px;">TA-006-FM</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Normal Goat Serum</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">ICN19135680</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Paraformaldehyde (PFA)</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">AAJ19943K2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Sterile Phosphate Buffered Saline (PBS)</td>
			<td style="width:196px;">ThermoFisher Scientific</td>
			<td style="width:166px;">14190250</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Triton-X 100</td>
			<td style="width:196px;">Sigma-Aldrich</td>
			<td style="width:166px;">T8787</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Supplies</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Adhesion Glass Slides</td>
			<td style="width:196px;">VWR</td>
			<td style="width:166px;">48311-703</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Desalting Columns</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">45-000-148</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Glass Cover Slips</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">12-544G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Hydrophobic Barrier Pen</td>
			<td style="width:196px;">Ted Pella</td>
			<td style="width:166px;">22311</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Microcentrifuge Tubes</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:166px;">05-402-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Slide Staining Tray</td>
			<td style="width:196px;">VWR</td>
			<td style="width:166px;">87000-136</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">Software</td>
			<td style="width:196px;"></td>
			<td style="width:166px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:295px;">FIJI</td>
			<td style="width:196px;">LOCI, UW-Madison.</td>
			<td style="width:166px;">Version 4.0</td>
			<td>https://fiji.sc/</td>
		</tr>
	</tbody>
</table>

in vivo immunofluorescence localization for assessment of therapeutic and diagnostic antibody biodistribution in cancer research

单克隆抗体 （mAbs） 是癌症检测、诊断和治疗的重要工具。它们用于解开蛋白质在肿瘤发生中的作用，可定向到癌症生物标志物，使肿瘤检测和表征，并可用于癌症治疗作为mAbs或抗体药物结合激活免疫效应细胞，以抑制信号通路，或直接杀死携带特定抗原的细胞。尽管在开发和生产新型和高度特异性的 mAbs 方面取得了临床进展，但肿瘤微环境的复杂性和异质性可能会损害诊断和治疗应用。因此，为了开发有效的基于抗体的疗法和诊断，评估基于抗体的偶联物与活肿瘤微环境的生物分布和相互作用至关重要。在这里，我们将 Vivo 免疫荧光定位 （IVIL） 描述为研究体内生理和病理条件下基于抗体的治疗和诊断的相互作用的新方法。在这项技术中，在体内静脉注射治疗或诊断抗原特异性抗体，并在隔离肿瘤中注射二级抗体。因此，IVIL反映了基于抗体的药物和靶向剂的体内生物分布。介绍了两种IVIL应用，用于评估基于抗体的造影剂在乳腺癌分子成像中的生物分布和可及性。该协议将允许未来的用户调整IVIL方法，以适应他们自己的基于抗体的研究应用。

IVIL 方法用于检查 B7-H3-ICG 和 Iso-ICG 的体内生物分布和组织相互作用，方法是允许制剂在静脉注射到活的动物体内后与目标组织相互作用 96 小时，然后一旦组织收获，在体外免疫染色过程中作为主要抗体。IVIL 方法还与 B7-H3 标记组织的标准体前 IF 染色进行了比较。正常乳腺不表示B7-H3标记，在体外确认，标准IF染色，不通过其他被动机制积累抗体，因此，未检测到B7-H3-ICG或...

Watch this Scientific Journal Video about In Vivo Immunofluorescence Localization for Assessment of Therapeutic and Diagnostic Antibody Biodistribution in Cancer Research at JoVE.com

In Vivo Immunofluorescence Localization for Assessment of Therapeutic and Diagnostic Antibody Biodistribution in Cancer Research

体内免疫荧光定位（IVIL）方法可用于利用体内肿瘤靶向和体外免疫染色的组合，在活体中检测抗体和抗体结合物在体内的生物体中的抗体和抗体结合方法。

在Vivo免疫荧光定位中用于评估癌症研究中的治疗和诊断抗体生物分布

Monoclonal antibodies (mAbs) are important tools in cancer detection, diagnosis, and treatment. They are used to unravel the role of proteins in ...

此活体免疫荧光定位方案（IVIL）评估了诊断和治疗抗体或抗体基制剂的体内生物分布，用于癌症研究、检测和治疗。与传统的免疫荧光染色（其中抗原的细胞表达在体内显示，IVIL方法阐明抗体和抗体为基础的制剂在活体动物中分布的方式。本视频将有助于了解 IVIL 方法的整体工作流。它显示了成功复制协议的其他应用和抗体的重要细节。观察小鼠从所需的癌症模型适当的肿瘤生长通过心或卡钳测量，然后再继续。净化兔子抗小鼠B7-H3和兔子IgG等型控制抗体在脱盐柱上，以去除防腐剂和储存缓冲液，按照制造商的指示。每种抗体的33微克的等量在单个微离心管中结合。按照文本协议中描述的麻醉后，为抗体溶液的尾静脉接种做好准备。用酒精擦拭三次，对动物的尾巴进行消毒。用热垫加热约30秒，使尾静脉扩张。避免加热整个动物。使用 27 量尾静脉导管，将蝴蝶针插入两个横向尾静脉之一。将血液回流可视化到导管中，以确保针头正确放置，使溶液完全进入动物，而不是被捕获在尾部。使用一块手术胶带小心地用插入的针头固定到舞台上的尾巴。用 25 微升无菌磷酸盐缓冲盐水冲洗导管。然后，使用胰岛素注射器将抗体溶液注射到导管中。使用 25 微升无菌 PBS 再次冲洗导管。从尾巴上取下针头，并施加压力以阻止任何出血。如文本协议所述，对鼠标进行人道安乐死，将鼠标放在超平位置。使用手术剪刀和钳子切除肿瘤组织。使用钳子只抓住最接近尾部的乳腺之间的皮肤外层，用一把手术剪刀做一个小切口。将封闭的剪刀放入切口中，然后慢慢打开尖端，小心地将皮肤与底层腹壁膜分开，保持其完好无损。在腹部进行垂直切口，继续将皮肤与内膜分开。在第三和第四乳腺之间，在腹部进行水平切口，允许皮肤缩回和乳腺的可视化。用钳子抓住每个肿瘤或正常腺体，用手术剪刀小心修剪附着的皮肤。将切除的组织放入组织一次性基模中，这些基模已预标记，并充满最佳的切割温度嵌入介质。将模具放在干冰上，快速冷冻模具。为了研究非靶向交付，切除其他感兴趣的组织或器官。使用低温恒温器，将 10 微米厚度的节块冷冻组织块，并放置相邻部分，将相邻截面放在预标记的粘附玻璃幻灯片上。用室温 PBS 冲洗冷冻组织幻灯片五分钟，以去除嵌入介质。使用疏水阻隔屏障笔划定组织部分，以减少染色过程中所需的溶液量。用4%的甲醛溶液修复组织部分5分钟。在 PBS 中冲洗幻灯片 5 分钟后，在 PBS 中用 0.5% Triton X-100 渗透组织部分 15 分钟。在 PBS 中再次冲洗幻灯片五分钟。在室温下，用PBS将含有每体积牛血清白蛋白3%的重量和每卷山羊血清5%体积的PBS块块组织一小时。在PBS中冲洗幻灯片五分钟后，用记录保持原抗体（如常见的核、血管或细胞质标记）孵育部分。在这里，大鼠防鼠CD31用于一到100稀释的阻解溶液。含有原抗体的幻灯片在四摄氏度下过夜，防止在滑动托盘上脱水。孵育后，在PBS中冲洗幻灯片五分钟三次。每次更改 PBS。用二次抗体孵育幻灯片，为初级抗体贴上标签。对于本应用，使用Alexa Fluor 546结合山羊抗兔抗体可视化抗B7-H3抗体，并可视化CD31与Alexa Fluor 488山羊抗大鼠二级抗体的阻断溶液。在室温下，保护幻灯片免受光线和脱水，在一小时。在 PBS 中像之前一样冲洗幻灯片后，将一滴安装介质放入组织切片的中心。小心放置盖玻片，避免夹住气泡。用清晰的指甲油密封盖玻片的边缘，并允许干燥。继续按照文本协议中所述进行共声显微镜成像和定量图像分析。代表性共合显显微图显示特定B7-H3抗体-ICG结合物和非特异性异型控制抗体-ICG结合定位在含有正常或癌组织乳腺的乳腺中。正常乳腺从动物静脉注射Iso-ICG或B7-H3-ICG和与CD31对等显示没有染色的抗体结合。正常组织通过标准外活体免疫荧光染色显示没有B7-H3的表达。在侵入性乳腺肿瘤中，B7-H3-ICG与体内第一接触点血管强烈结合，然后能够从血管外体异质染色肿瘤上皮。Iso-ICG 显示肿瘤组织内的非特异性积累。标准外体免疫荧光染色显示上皮细胞和内皮细胞上B7-H3标记的均匀表达。代表性共体显微图显示体内免疫荧光定位方法，用于检测乳腺癌和正常乳腺中的网蛋白-1表达或等型控制表达。体内免疫荧光定位确认MMTV-PyMT肿瘤中网林-1的上皮信号，但在正常乳腺中不确认。此外，在乳腺肿瘤的内皮细胞上还有一个强烈的网林-1信号，如结黄信号所示。正常乳腺中的信号明显较弱。IVIL方法需要优化抗体剂量、组织采集时间、二次抗体稀释等方法，才能成功实施。IVIL 允许定位使用标准免疫荧光染色无法检测到的构象表位，这需要组织处理。此外，还可以收集带肿瘤小鼠的健康器官，以评估治疗抗体的靶外传递。随着抗体治疗和对比剂在癌症研究和管理中的使用增加，IVIL方法高度适用于药物研发。癌症治疗、分子成像、炎症和其他领域的研究人员发现，IVIL方法提供了有用的信息。

Research

JoVE Journal

Cancer Research

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

该<em&gt;果蝇</em&gt;成虫盘瘤模型：基因的表达与肿瘤侵袭性的可视化和定量遗传马赛克

Drosophila melanogaster has emerged as a powerful experimental system for functional and mechanistic studies of tumor development and progression in the ...

科研

癌症研究

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

体内在肿瘤微环境中 pH 值、 pO2、氧化还原状态、磷酸盐和谷胱甘肽浓度的 EPR 评价

This protocol demonstrates the capability of low-field electron paramagnetic resonance (EPR)-based techniques in combination with functional paramagnetic ...

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

用抗体阵列对信号转导通路的审问对酪氨酸激酶抑制剂的抗性评价

Cancer patients with an aberrant regulation of the protein phosphorylation networks are often treated with the tyrosine kinase inhibitors. Response rates ...

Using 22C3 Anti-PD-L1 Antibody Concentrate on Biopsy and Cytology Samples from Non-small Cell Lung Cancer Patients

22C3 Anti-PD-L1 抗体精矿在非小细胞肺癌活检及细胞学标本中的应用

Pembrolizumab monotherapy has been approved for the first- and second-line treatment of patients with PD-L1-expressing advanced non-small cell lung cancer ...

Establishment of Gastric Cancer Patient-derived Xenograft Models and Primary Cell Lines

建立胃癌患者衍生异种移植模型和原细胞系

The use of preclinical models to advance our understanding of tumor biology and investigate the efficacy of therapeutic agents is key to cancer research. ...

Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence

免疫荧光对非小细胞肺癌患者循环肿瘤细胞的半自动PD-L1特征化及枚举

Circulating tumor cells (CTCs) derived from the primary tumor are shed into the bloodstream or lymphatic system. These rare cells (1&#8722;10 cells per mL ...

Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody

靶性抗原特异性治疗抗体的肿瘤及组织分布分析

Monoclonal antibodies are high affinity multifunctional drugs that work by variable independent mechanisms to eliminate cancer cells. Over the last few ...

Enhanced Viability for Ex vivo 3D Hydrogel Cultures of Patient-Derived Xenografts in a Perfused Microfluidic Platform

在全融合微流体平台中增强患者衍生异种移植物的外体 3D 水凝胶培养物的生存能力

Patient-derived xenografts (PDX), generated when resected patient tumor tissue is engrafted directly into immunocompromised mice, remain biologically ...

Immunofluorescence Imaging of DNA Damage and Repair Foci in Human Colon Cancer Cells

人类结肠癌细胞DNA损伤与修复福西的免疫荧光成像

The purpose of the manuscript is to provide a step-by-step protocol for performing immunofluorescence microscopy to study the radiation-induced DNA damage ...