SIMPLE: A Sequential Immunoperoxidase Labeling and Erasing Method
SIMPLE: A Sequential Immunoperoxidase Labeling and Erasing Method
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DOI：
10.1369/jhc.2009.953612
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发表时间：
2009-10-01
影响因子：
3.2
通讯作者：
Mandell, James W.
中科院分区：
文献类型：
--
作者：
Glass, George;Papin, Jason A.;Mandell, James W.
能够同时观察细胞和组织中多种抗原的表达可为细胞生物学和机体生物学提供有力的见解。然而，标准方法仅限于同时使用两三种探针，并且尚未在石蜡包埋组织的常规使用中被广泛采用。我们开发了一种名为顺序免疫过氧化物酶标记和消除（SIMPLE）的新方法，该方法能够在单个组织切片中同时观察至少五种标记物。利用醇溶性过氧化物酶底物3 - 氨基 - 9 - 乙基咔唑，并结合一种用于抗体 - 抗原解离的快速无损方法，我们证明了在保留组织抗原性以进行重复标记的同时消除单次免疫组化染色结果的能力。使用全玻片扫描仪极大地促进了SIMPLE方法的应用，该扫描仪能够捕获每次顺序染色的结果而不会丢失任何信息。（《组织化学与细胞化学杂志》2009年第57卷第899 - 905页）
The ability to simultaneously visualize expression of multiple antigens in cells and tissues can provide powerful insights into cellular and organismal biology. However, standard methods are limited to the use of just two or three simultaneous probes and have not been widely adopted for routine use in paraffin-embedded tissue. We have developed a novel approach called sequential immunoperoxidase labeling and erasing (SIMPLE) that enables the simultaneous visualization of at least five markers within a single tissue section. Utilizing the alcohol-soluble peroxidase substrate 3-amino-9-ethylcarbazole, combined with a rapid non-destructive method for antibody-antigen dissociation, we demonstrate the ability to erase the results of a single immunohistochemical stain while preserving tissue antigenicity for repeated rounds of labeling. SIMPLE is greatly facilitated by the use of a whole-slide scanner, which can capture the results of each sequential stain without any information loss. (J Histochem Cytochem 57:899-905, 2009)