In Vivo and Ex Vivo Analysis of Omental Adhesion in Ovarian Cancer.
In Vivo and Ex Vivo Analysis of Omental Adhesion in Ovarian Cancer.
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DOI：
10.1007/978-1-0716-1956-8_14
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发表时间：
2022-01-01
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影响因子：
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通讯作者：
Hilliard, Tyvette S
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作者：
Harper, Elizabeth I;Hilliard, Tyvette S
在体内和离体分析网膜粘连在卵巢癌（OvCa）是必要的了解OvCa转移的动力学。在这里，我们描述的方法来分析OvCa网膜粘连，包括在体内和体外荧光成像，先进的显微镜和组织学技术。荧光标记的OvCa细胞的使用允许网膜肿瘤可视化和定量的粘附和肿瘤研究。此外，先进的显微镜模式允许OvCa网膜粘连的可视化和多路复用分析。二次谐波显微镜允许网膜胶原蛋白的可视化和分析，特别是随着肿瘤进展形成的肿瘤相关胶原蛋白特征。扫描电子显微镜用于观察OvCa细胞和大网膜之间的微观细节，例如隧道纳米管或微绒毛。组织学方法用于研究几种肿瘤内性质，包括使用苏木精和伊红染色使肿瘤结构可视化;用Masson三色染色定量胶原蛋白;用胶原蛋白杂交肽分析胶原蛋白结构;以及用免疫组织化学鉴定许多标志物，包括但不限于增殖、免疫细胞类型、粘附分子和成纤维细胞。体内和离体成像模式以及随后的分析都可以提供对转移性OvCa细胞和网膜的相互作用的洞察。
In vivo and ex vivo analyses of omental adhesion in ovarian cancer (OvCa) are necessary to understand the dynamics of OvCa metastasis. Here we describe methods to analyze OvCa omental adhesion, including in vivo and ex vivo fluorescent imaging, advanced microscopy, and histological techniques. The use of fluorescently tagged OvCa cells allows for omental tumor visualization and quantification in adhesion and tumor studies. Additionally, advanced microscopy modalities allow for visualization and multiplexed analysis of OvCa omental adhesion. Second harmonic generation microscopy permits the visualization and analysis of omental collagen, specifically the tumor-associated collagen signature that forms as the tumor progresses. Scanning electron microscopy is used for the observation of microscopic details between OvCa cells and the omentum, such as tunneling nanotubes or microvilli. Histological methods are used to investigate several intratumoral properties including visualizing tumor structure using hematoxylin and eosin stain; quantifying collagen with Masson's trichrome stain; analyzing collagen structure with a collagen hybridizing peptide; and identifying a number of markers including, but not limited to proliferation, immune cell types, adhesion molecules, and fibroblasts with immunohistochemistry. Both the in vivo and ex vivo imaging modalities and subsequent analysis can provide insight into the interaction of metastasizing OvCa cells and the omentum.