Consecutive Immunohistochemical Staining on a Single Slide is Effective in Confirming Melanocytic Derivation
Consecutive Immunohistochemical Staining on a Single Slide is Effective in Confirming Melanocytic Derivation
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DOI：
10.1097/pai.0000000000000665
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发表时间：
2020-10-01
影响因子：
1.6
通讯作者：
Shalin, Sara C.
中科院分区：
文献类型：
--
作者：
Campbell, Katelynn K.;Gardner, Jerad M.;Shalin, Sara C.
背景：前哨淋巴结转移性黑色素瘤通常难以仅凭形态学发现。根据美国癌症联合委员会的分期指南，淋巴结中单个非典型黑素细胞符合转移的条件，无论是通过形态学还是免疫组织化学鉴定，但单个细胞染色必须令人信服。我们建议，在单个玻片上进行第二次免疫组织化学检查将允许更有信心地诊断微转移。材料和方法：我们设计了一项技术研究，以确定在先前染色的玻片上应用第二抗体是否能成功检测相同数量的细胞。黑素细胞肿瘤用Ventana Benchmark Ultra染色机行SOX-10染色，盖玻片检查，然后盖玻片去除并应用MART-1(Ventana A103)。抗体应用和染色试剂盒(AP-RED和DAB)的顺序颠倒，以建立该方案的可靠性和健壮性。结果：所有黑素细胞用SOX-10和MART-1标记，并根据染色顺序产生不同的染色质量，并使用染色试剂盒。最佳组合是先施红豆-1，再施棕袜-10。结论：用SOX-10和MART-1对黑素细胞进行连续染色可以提高黑素细胞鉴定的诊断置信度，特别是在检测单细胞、前哨淋巴结微转移或双重免疫组织化学染色可能不可用的情况下。
Background: Metastatic melanoma in sentinel lymph nodes is often elusive to detect with morphology alone. Per American Joint Committee on Cancer staging guidelines, a single atypical melanocyte in lymph node qualifies as metastasis, whether identified by morphology or immunohistochemistry, but single cell staining must be convincing. We propose that the use of a second immunohistochemical run performed on a single slide will allow for more confident diagnosis of micrometastases. Materials and Methods: We designed a technical study to determine whether a second antibody application on previously stained slides can successfully detect the same population of cells. Melanocytic neoplasms were stained with SOX-10 using Ventana Benchmark Ultra stainers, coverslipped, and examined, followed by coverslip removal and application of MART-1 (Ventana A103). The order of antibody application and chromagen detection kit (AP-RED vs. DAB) was reversed to establish reliability and robustness of the protocol. Results: All melanocytes marked with SOX-10 and MART-1, and produced a range of staining quality that varied based on order of stain application and chromagen kit were used. The optimal combination was red MART-1 applied first followed by brown SOX-10 applied second. Conclusions: Consecutive staining of melanocytes with SOX-10 and MART-1 may improve diagnostic confidence of melanocyte identification, particularly in detection of single cell, micrometastases in sentinel lymph nodes or in situations where dual immunohistochemical stains may be unavailable.