Name: expanding the comprehension of the tumor microenvironment using mass spectrometry imaging of formalin-fixed and paraffin-embedded tissue samples
Uploaded: 2022-06-29T13:00:00
Description: Watch this Scientific Journal Video about Expanding the Comprehension of the Tumor Microenvironment using Mass Spectrometry Imaging of Formalin-Fixed and Paraffin-Embedded Tissue Samples at JoVE.com

作者感谢来自编辑服务，MD安德森研究医学图书馆编辑服务的Don Norwood编辑本文以及MD安德森翻译分子病理学系的多重免疫荧光和图像分析实验室。该出版物的部分结果来自通过美国国家癌症研究所（NCI）合作协议（U24CA224285）向德克萨斯大学MD安德森癌症中心癌症免疫监测和分析中心（CIMAC）提供的癌症免疫监测和分析中心 - 癌症免疫数据共享网络（CIMAC-CIDC）的科学和财政支持所促进的研究。

根据德克萨斯大学MD安德森癌症中心的机构审查委员会，出于研究目的获得了组织样本，并且样本进一步去鉴定。
1. 抗体选择
<ol><li>定义研究问题以选择最佳可用的抗体克隆。使用人类蛋白质图谱、已发表的研究和抗体制造商网站作为研究资源。 注意：选择足够的人体组织质控品和相同的人体组织质控品在不同步骤中染色对于确保标记物在正确的位置和正确?...

<ol>
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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computational+approaches+for+characterizing+the+tumor+immune+microenvironment.">Computational approaches for characterizing the tumor immune microenvironment.</a> Immunology. 158 (2), 70-84 (2019).</li><li>Eisenstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+sorting:+Divide+and+conquer.">Cell sorting: Divide and conquer.</a> Nature. 441 (7097), 1179-1185 (2006).</li><li>Scognamiglio, G., 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=Multiplex+immunohistochemistry+assay+to+evaluate+the+melanoma+tumor+microenvironment.">Multiplex immunohistochemistry assay to evaluate the melanoma tumor microenvironment.</a> Methods in Enzymology. 635, 21-31 (2020).</li><li>Guo, 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=A+34-marker+panel+for+imaging+mass+cytometric+analysis+of+human+snap-frozen+tissue.">A 34-marker panel for imaging mass cytometric analysis of human snap-frozen tissue.</a> Frontiers in Immunology. 11, 1466 (2020).</li><li>Fu, T., 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=Spatial+architecture+of+the+immune+microenvironment+orchestrates+tumor+immunity+and+therapeutic+response.">Spatial architecture of the immune microenvironment orchestrates tumor immunity and therapeutic response.</a> Journal of Hematology &amp; Oncology. 14 (1), 98 (2021).</li><li>Rost, 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=Multiplexed+ion+beam+imaging+analysis+for+quantitation+of+protein+expresssion+in+cancer+tissue+sections.">Multiplexed ion beam imaging analysis for quantitation of protein expresssion in cancer tissue sections.</a> Laboratory Investigation. 97 (8), 992-1003 (2017).</li><li>Keren, L., 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+structured+tumor-immune+microenvironment+in+triple+negative+breast+cancer+revealed+by+multiplexed+ion+beam+imaging.">A structured tumor-immune microenvironment in triple negative breast cancer revealed by multiplexed ion beam imaging.</a> Cell. 174 (6), 1373-1387 (2018).</li><li>Mathieson, W., Thomas, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+formalin-fixed,+paraffin-embedded+biospecimens+must+be+used+in+genomic+medicine:+An+evidence-based+review+and+conclusion.">Why formalin-fixed, paraffin-embedded biospecimens must be used in genomic medicine: An evidence-based review and conclusion.</a> The Journal of Histochemistry and Cytochemistry. 68 (8), 543-552 (2020).</li><li>Baghban, R., 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=Tumor+microenvironment+complexity+and+therapeutic+implications+at+a+glance.">Tumor microenvironment complexity and therapeutic implications at a glance.</a> Cell Communication and Signaling. 18 (1), 59 (2020).</li><li>Cai, S., Allam, M., Coskun, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplex+spatial+bioimaging+for+combination+therapy+design.">Multiplex spatial bioimaging for combination therapy design.</a> Trends in Cancer. 6 (10), 813-818 (2020).</li><li>Allam, M., Cai, S., Coskun, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplex+bioimaging+of+single-cell+spatial+profiles+for+precision+cancer+diagnostics+and+therapeutics.">Multiplex bioimaging of single-cell spatial profiles for precision cancer diagnostics and therapeutics.</a> NPJ Precision Oncology. 4, 11 (2020).</li><li>Rad, H. 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=The+Pandora's+box+of+novel+technologies+that+may+revolutionize+lung+cancer.">The Pandora's box of novel technologies that may revolutionize lung cancer.</a> Lung Cancer. 159, 34-41 (2021).</li><li>Tan, W. C. C., 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=Overview+of+multiplex+immunohistochemistry/immunofluorescence+techniques+in+the+era+of+cancer+immunotherapy.">Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy.</a> Cancer Commununications. 40 (4), 135-153 (2020).</li><li>Ptacek, 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=Multiplexed+ion+beam+imaging+(MIBI)+for+characterization+of+the+tumor+microenvironment+across+tumor+types.">Multiplexed ion beam imaging (MIBI) for characterization of the tumor microenvironment across tumor types.</a> Laboratory Investigation. 100 (8), 1111-1123 (2020).</li></ol>

全面阐明TME多个组成部分之间复杂而复杂的相互作用仍然是癌症研究的关键目标。作为这项工作的一部分，制造商已经推出了许多多重检测方法，特别是在过去的5年中。多重空间分析是一种多功能且功能强大的工具，有助于同时对多个靶标进行分类，同时保持肿瘤样品中的结构形态。空间分析技术可以使用荧光成像或质谱法进行，例如本文11、12、<...

克隆性肿瘤群体周围众多细胞类型的重要性是癌变分类的关键因素。在建立基于免疫的疗法作为癌症治疗武器库的一部分之后，对阐明这种肿瘤微环境（TME）组成和相互作用的兴趣不断上升。因此，治疗策略已从以肿瘤为中心的方法转变为以TME为中心的方法1。
近年来，阐明免疫细胞在肿瘤监测和癌症发展中的作用的努力急剧增加2，3.在医学研究中，大量的方法，包括基于细胞术的方法以及单重和多重成像技术，作为破译TME多种元素独特相互作用的尝试的一部分， 出现了。
流式细胞术（20世纪60年代发明）、荧光活化细胞分选和质量细胞术等开创性方法主要集中在鉴定和定量TME组分4上。尽管基于细胞术的定量技术允许免疫景观表型分析，但确定细胞空间分布是不可能的。相反，诸如标准单重免疫组织化学之类的方法保留了组织结构并使研究人员能够分析细胞分布，尽管单个组织切片中靶标数量的减少是这些方法的局限性5，6。在过去几年中，用于单细胞分辨率的多重成像技术（如多重免疫荧光，条形码荧光成像和成像质谱）已成为使用相同的组织切片7获取同时标记染色信息的综合策略。
在这里，我们介绍了一种将金属标记抗体和二次离子质谱耦合的技术，并使用福尔马林固定，石蜡包埋（FFPE）和新鲜冷冻（FF）组织样品8，9实现单细胞分辨率定量，标记物共表达（表型）和空间分析。FFPE样品是用于组织存档样品的最广泛使用的材料，并且比新鲜冷冻样品10更容易获得多重成像技术。此外，该技术还提供了在几个月后重新获取图像的可能性。在这里，我们讨论了使用FFPE组织样品的染色和图像处理方案。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>100% Reagent Alcohol</td>
			<td>Sigma-Aldrich</td>
			<td>R8382</td>
			<td></td>
		</tr>
		<tr>
			<td>95% Reagent Alcohol</td>
			<td>Sigma-Aldrich</td>
			<td>R3404</td>
			<td></td>
		</tr>
		<tr>
			<td>80% Reagent Alcohol</td>
			<td>Sigma-Aldrich</td>
			<td>R3279</td>
			<td></td>
		</tr>
		<tr>
			<td>70% Reagent Alcohol</td>
			<td>Sigma-Aldrich</td>
			<td>R315</td>
			<td></td>
		</tr>
		<tr>
			<td>20X TBS-T</td>
			<td>Ionpath</td>
			<td>567005</td>
			<td></td>
		</tr>
		<tr>
			<td>10X Low-Barium PBS pH 7.4</td>
			<td>Ionpath</td>
			<td>567004</td>
			<td></td>
		</tr>
		<tr>
			<td>10X Tris pH 8.5&#160;</td>
			<td>Ionpath</td>
			<td>567003</td>
			<td></td>
		</tr>
		<tr>
			<td>4&#176;C Refrigerator</td>
			<td>ThermoScientific</td>
			<td>REVCO</td>
			<td></td>
		</tr>
		<tr>
			<td>Aerosol Barrier Pipette Tips P10</td>
			<td>Olympus</td>
			<td>24-401</td>
			<td></td>
		</tr>
		<tr>
			<td>Aerosol Barrier Pipette Tips P20</td>
			<td>Olympus</td>
			<td>24-404</td>
			<td></td>
		</tr>
		<tr>
			<td>Aerosol Barrier Pipette Tips P200</td>
			<td>Olympus</td>
			<td>24-412</td>
			<td></td>
		</tr>
		<tr>
			<td>Aerosol Barrier Pipette Tips P1000</td>
			<td>Olympus</td>
			<td>24-430</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifugal Filter Ultrafree-MC</td>
			<td>Fisher Scientific</td>
			<td>UFC30VV00</td>
			<td></td>
		</tr>
		<tr>
			<td>Deionized H2O</td>
			<td>Ionpath</td>
			<td>567002</td>
			<td></td>
		</tr>
		<tr>
			<td>Donkey serum</td>
			<td>Sigma-Aldrich</td>
			<td>D9663</td>
			<td></td>
		</tr>
		<tr>
			<td>EasyDip Slide Staining Jar, Green</td>
			<td>Electron Microscopy Sciences</td>
			<td>71385-G</td>
			<td></td>
		</tr>
		<tr>
			<td>EasyDip Slide Staining Jar, Yellow</td>
			<td>Electron Microscopy Sciences</td>
			<td>71385-Y</td>
			<td></td>
		</tr>
		<tr>
			<td>EasyDip Slide Staining Kit (Jar+Rack), White</td>
			<td>Electron Microscopy Sciences</td>
			<td>71388-01</td>
			<td></td>
		</tr>
		<tr>
			<td>EasyDip Stainless Steel Holder</td>
			<td>Electron Microscopy Sciences</td>
			<td>71388-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Glutaraldehyde 70% EM Grade</td>
			<td>Electron Microscopy Sciences</td>
			<td>16360</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat Induced Epitope Retrieval (HIER) buffer: 10X Tris with EDTA, pH 9</td>
			<td>Dako</td>
			<td>S2367</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat resistant slide chamber</td>
			<td>Electron Microscopy Sciences</td>
			<td>62705-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrophobic barrier pen</td>
			<td>Fisher</td>
			<td>50-550-221</td>
			<td></td>
		</tr>
		<tr>
			<td>MIBI/O software</td>
			<td>Ionpath</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>MIBIcontrol software</td>
			<td>Ionpath</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>MIBIslide</td>
			<td>Ionpath</td>
			<td>567001</td>
			<td></td>
		</tr>
		<tr>
			<td>MIBIscope</td>
			<td>Ionpath</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Microcentrifuge</td>
			<td>Eppendorf</td>
			<td>5415D</td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Leica</td>
			<td>RM2135</td>
			<td></td>
		</tr>
		<tr>
			<td>Moisture Chamber (Humid Chamber)</td>
			<td>Simport</td>
			<td>M922-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (PBS) Tablets</td>
			<td>Fisher Scientific</td>
			<td>BP2944100</td>
			<td></td>
		</tr>
		<tr>
			<td>PT Module</td>
			<td>Thermo Scientific</td>
			<td>A80400012</td>
			<td></td>
		</tr>
		<tr>
			<td>Rapid-Flow Sterile Disposable Filter Units</td>
			<td>Fisher Scientific</td>
			<td>097403A</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaker</td>
			<td>BioRocker</td>
			<td>S2025</td>
			<td></td>
		</tr>
		<tr>
			<td>Spin column (Ultrafree-MC Spin Filter, 0.5mL 0.1&#956;m )</td>
			<td>MillQ</td>
			<td>UFC30VV00</td>
			<td></td>
		</tr>
		<tr>
			<td>Slide oven</td>
			<td>Fisher Scientific</td>
			<td>6901</td>
			<td></td>
		</tr>
		<tr>
			<td>Vaccum Cabinet Desiccator</td>
			<td>VWR</td>
			<td>30621-076</td>
			<td></td>
		</tr>
		<tr>
			<td>Task-whipe</td>
			<td>Kimberly Clark</td>
			<td>34155</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Sigma-Aldrich</td>
			<td>534056-4L</td>
			<td></td>
		</tr>
	</tbody>
</table>

expanding the comprehension of the tumor microenvironment using mass spectrometry imaging of formalin-fixed and paraffin-embedded tissue samples

基于免疫的疗法的进步彻底改变了癌症治疗和研究。这引发了对肿瘤免疫景观表征的需求不断增长。虽然标准免疫组化适用于研究组织结构，但它仅限于分析少量标记物。相反，流式细胞术等技术可以同时评估多个标志物，尽管有关组织形态的信息丢失。近年来，整合表型和空间分析的多重策略已成为表征肿瘤免疫景观的综合方法。 在这里，我们讨论了一种结合金属标记抗体和二次离子质谱的创新技术，重点介绍了测定开发和优化，组织制备以及图像采集和处理中的技术步骤。在染色之前，必须开发和优化金属标记的抗体组合。这种高复级图像系统在单个组织切片中支持多达40种金属标记的抗体。值得注意的是，信号干扰的风险随着面板中包含的标记数量而增加。在设计面板后，应特别注意对抗体的金属同位素分配，以尽量减少这种干扰。使用一小部分抗体进行初步的面板测试，随后在对照组织中对整个面板进行测试。获得福尔马林固定的石蜡包埋组织切片并将其安装在镀金载玻片上并进一步染色。染色需要2天，与标准免疫组化染色非常相似。一旦样品被染色，它们就被放置在图像采集仪器中。选择视野，获取、上传和存储图像。最后阶段是图像准备，以便使用系统的图像处理软件进行滤波和消除干扰。该平台的缺点是缺乏分析软件。但是，生成的图像由不同的计算病理学软件支持。

获得扁桃体和肺腺癌TMA组织切片（5mm厚）并按照有关切片的组织大小和安全边缘的规格放置在金涂层载玻片的中间。组织边缘与载玻片的侧缘和下边界之间分别有5 mm和10 mm的自由玻璃边缘对于最佳染色是必需的。在染色之前，将组织切片在烤箱中烘烤过夜，以确保切片正确粘附在载玻片上。抗体组合由23个标记物组成。在同一染色批次中，包括扁桃体载玻片和含有多个组织的TMA载玻片，以评估肺?...

Watch this Scientific Journal Video about Expanding the Comprehension of the Tumor Microenvironment using Mass Spectrometry Imaging of Formalin-Fixed and Paraffin-Embedded Tissue Samples at JoVE.com

Expanding the Comprehension of the Tumor Microenvironment using Mass Spectrometry Imaging of Formalin-Fixed and Paraffin-Embedded Tissue Samples

在癌症免疫治疗时代，人们对阐明肿瘤微环境动力学的兴趣惊人地增加。该协议详细介绍了质谱成像技术及其染色和成像步骤，允许高度多路复用的空间分析。

使用福尔马林固定和石蜡包埋组织样品的质谱成像扩展肿瘤微环境的理解

Advances in immune-based therapies have revolutionized cancer treatment and research. This has triggered growing demand for the characterization of the ...

这些方案能够使用福尔马林固定和石蜡包埋和新鲜冷冻组织样品进行单细胞分辨率定量、标记共表达和特殊分析，以阐明免疫细胞在癌症中的作用。该技术是一种强大的工具，有助于在单个组织切片上同时表征40个标记物，广泛保留样品中的组织结构。关键步骤是抗体优化和验证。未能完成该步骤，可能会导致低质量的图像，从而可能损害保留高质量的可重现数据和结果。首先将所有含有抗体的试管以10， 000 x G旋转五分钟，将单个抗体添加到封闭缓冲液中并充分混合。移液时不要打扰抗体管的底部。通过用100微升封闭缓冲液预润湿0.1微米离心过滤装置来过滤抗体组合。以 10， 000 x G 旋转过滤器两分钟，然后用移液器去除阻塞缓冲液流出物。然后将抗体组合转移到离心柱中，并以10， 000 x G旋转过滤器两分钟。丢弃离心柱，使用流通物作为过滤的抗体组合。对于抗体染色，通过将每张载玻片侧倾并在任务刮水器上轻轻敲击边缘，从载玻片上去除封闭缓冲液。将载玻片放入水分室中，并向组织中加入100至200微升抗体预混液。将阳性和阴性对照载玻片添加到染色批次中，并将载玻片在4摄氏度下孵育过夜。要设置仪器，请登录到控制软件，如果仪器处于睡眠模式，请单击唤醒。单击“交换样品”以加载幻灯片，然后单击“继续”以打开门。将载玻片放入装载槽中，样品朝上，标签在右侧，单击“继续”关闭门。选择项目，然后选择加载示例的幻灯片名称。在幻灯片光学图像窗格上可视化全景图像以进行视野或FOV选择，单击模式菜单并选择二次电子检测器，然后单击成像模式菜单并选择QC 300微米。单击“点动阶段”以控制 FOV 位置，然后单击箭头以导航并选择 FOV 的位置。单击添加 FOV，将 400 微米 x 400 微米设置为视场大小，并选择精细作为成像模式和 1 毫秒的停留时间，单击确认。创建 FOV 列表后，继续图像焦点并通过将光束移动到不会成像的组织区域来调整柱头。调整参数，直到获得清晰的中心图像，单击开始运行。采集的图像将自动上传并存储在基于Web的图像管理应用程序中。此处显示的矩阵描述了从探针纯度和氧化物得出的串扰百分比。这里的蓝色框表示大于或等于 0.5% 串扰，透明框表示小于 0.5% 串扰。对于 Mass 标签，此处显示了对无通道、一个或两个通道或两个以上通道贡献至少 0.5% 串扰的探头。还显示了从无探头、一个或两个探头或两个以上探头接收至少 0.5% 串扰的质量通道。这里描绘了肺腺癌、非肿瘤性肾脏和扁桃体等不同组织中的代表性染色图像。这里展示了使用该技术染色的扁桃体组织切片的图像，并在过滤和校正之前使用第三方数字图像分析软件程序进行可视化。此处显示了经过过滤和校正步骤后相同条件下同一部分的图像。该方法生成的图像可以上传到第三方数字图像分析软件，可以进行组织区室化、细胞分割、标记共表达以及邻域分析和远距离分析等综合分析。通过研究可以获得高度多重成像方法。通过构建不同的面板，研究人员可以研究癌症，正常，副肿瘤，炎症和感染性样本。

Research

JoVE Journal

Cancer Research

Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples

档案福尔马林固定，石蜡包埋的人类口咽鳞状细胞癌样品中T细胞亚群的四色荧光免疫组织化学

The four-color fluorescence immunohistochemistry (IHC) technique is a method to quantify cell populations of interest while taking into account their ...

科研

癌症研究

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

二维凝胶电泳结合质谱法分析人垂体腺瘤组织蛋白质组

Human pituitary adenoma (PA) is a common tumor that occurs in the human pituitary gland in the hypothalamus-pituitary-targeted organ axis systems, and may ...

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

磷酸浓缩结合无标签定量质谱法研究前列腺癌的 Phosphoproteome

Phosphoproteomics involves the large-scale study of phosphorylated proteins. Protein phosphorylation is a critical step in many signal transduction ...

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

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 ...

Automated Multiplex Immunofluorescence Panel for Immuno-oncology Studies on Formalin-fixed Carcinoma Tissue Specimens

福尔马林固定癌组织标本免疫肿瘤学自动复合免疫荧光小组研究

Continued developments in immuno-oncology require an increased understanding of the mechanisms of cancer immunology. The immunoprofiling analysis of ...

Measuring Bone Remodeling and Recreating the Tumor-Bone Microenvironment Using Calvaria Co-culture and Histomorphometry

利用卡尔瓦里亚共同培养和教条法测量骨重塑和再造肿瘤-骨微环境

Bone is a connective tissue constituted of osteoblasts, osteocytes, and osteoclasts and a mineralized extracellular matrix, which gives it its strength ...

Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry

利用成像质谱技术捕获组织与细胞之间的小分子通信

Imaging mass spectrometry (IMS) has routinely been applied to three types of samples: tissue sections, spheroids, and microbial colonies. These sample ...

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

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 ...

Pathological Analysis of Lung Metastasis Following Lateral Tail-Vein Injection of Tumor Cells

肿瘤细胞外侧尾静脉注射后肺转移的病理分析

Metastasis, the primary cause of morbidity and mortality for most cancer patients, can be challenging to model preclinically in mice. Few spontaneous ...

Isolation of Proximal Fluids to Investigate the Tumor Microenvironment of Pancreatic Adenocarcinoma

分离近端液研究胰腺腺癌肿瘤微环境

Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death, and soon to become the second. There is an urgent need of variables ...