基于超敏半抗原的多重荧光免疫组化分析。
UltraPlex Hapten-Based Multiplexed Fluorescent Immunohistochemistry.
机构信息
Cell IDx, San Diego, CA, USA.
Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, USA.
出版信息
Methods Mol Biol. 2021;2350:267-287. doi: 10.1007/978-1-0716-1593-5_17.
The UltraPlex method for multiplexed two-dimensional fluorescent immunohistochemistry is described, in which hapten tags conjugated to primary antibodies facilitate multiplexed imaging of four or more antigens per tissue section at once. Anti-hapten secondary antibodies labeled with fluorophores provide amplified signal for detection, which is accomplished using a standard fluorescent microscope or digital slide scanner. The protocol is rapid and straightforward and utilizes conventionally prepared tissue samples. The resulting staining is highly sensitive and specific, enabling high-resolution imaging of multiple cellular subtypes within tissue samples. Tumor cells and tumor-infiltrating lymphocytes are presented as examples. Multiple 4-plex-stained tissue samples can be digitally overlaid to create 8-plex (or more) high-content images, enabling visualization of distribution of complex cellular subtypes across tissues.
UltraPlex 方法用于多重二维荧光免疫组织化学，该方法通过将连接半抗原的一抗用于组织切片，可同时对 4 种或更多种抗原进行多重成像。荧光标记的抗半抗原二抗提供了增强的信号检测，该检测可使用标准荧光显微镜或数字切片扫描仪完成。该方案快速而直接，并且利用常规制备的组织样本。得到的染色具有高度的敏感性和特异性，能够在组织样本中对多种细胞亚型进行高分辨率成像。本文以肿瘤细胞和肿瘤浸润淋巴细胞为例进行介绍。可以对多个 4 重染色的组织样本进行数字叠加，以创建 8 重（或更多）高内涵图像，从而可视化复杂细胞亚型在组织中的分布。
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