Multicolor-FICTION -: Expanding the possibilities of combined morphologic, immunophenotypic, and genetic single cell analyses
Multicolor-FICTION -: Expanding the possibilities of combined morphologic, immunophenotypic, and genetic single cell analyses
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DOI：
10.1016/s0002-9440(10)64197-1
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发表时间：
2002-08-01
影响因子：
6
通讯作者：
Siebert, R
中科院分区：
文献类型：
--
作者：
Martín-Subero, JI;Chudoba, I;Siebert, R
细胞的表型和基因型分析对于理解发病机制以及诊断和分类恶性肿瘤和其他疾病越来越重要。我们报告了一种基于FICTION（荧光免疫表型分析和间期细胞遗传学作为肿瘤研究工具）技术的新方法，该方法能够同时检测单细胞的形态学、免疫表型和遗传特征。作为先决条件，B细胞非霍奇金淋巴瘤和间变性大细胞淋巴瘤的荧光原位杂交检测已被开发。这些检测试剂盒可同时检测这些肿瘤中最常见的原发性染色体畸变，如t（8;14）、t（11;14）、t（14;18）和t（3;14），以及ALK基因的各种重排。为了建立ELISA技术，将这些测定与谱系或肿瘤特异性抗原（即分别为CD 20和ALK）的免疫表型检测相结合。为了评价多焦平面成像实验，通过多焦平面的自动顺序捕获进行图像采集。因此，获得了三维信息。将该检测应用于充分表征的淋巴瘤样本，证明了该技术的性能、有效性和诊断能力。未来的诊断应用包括肿瘤前病变、早期和微小残留疾病或微转移的检测。
Phenotypic and genotypic analyses of cells are increasingly essential for understanding pathogenetic mechanisms as well as for diagnosing and classifying malignancies and other diseases. We report a novel multicolor approach based on the FICTION (fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms) technique, which enables the simultaneous detection of morphological, immunophenotypic, and genetic characteristics of single cells. As prerequisite, multicolor interphase fluorescence in situ hybridization assays for B-cell non-Hodgkin's lymphoma and anaplastic large-cell lymphoma have been developed. These assays allow the simultaneous detection of the most frequent primary chromosomal aberrations in these neoplasms, such as t(8;14), t(11;14), t(14;18), and t(3;14), and the various rearrangements of the ALK gene, respectively. To establish the multicolor FICTION technique, these assays were combined with the immunophenotypic detection of lineage- or tumor-specific antigens, namely CD20 and ALK, respectively. For evaluation of multicolor FICTION experiments, image acquisition was performed by automatic sequential capturing of multiple focal planes. Thus, three-dimensional information was obtained. The multicolor FICTION assays were applied to well-characterized lymphoma samples, proving the performance, validity, and diagnostic power of the technique. Future multicolor FICTION applications include the detection of preneoplastic lesions, early stage and minimal residual diseases, or micrometastases.