In-situ-topoproteome analysis of cutaneous lymphomas: Perspectives of assistance for dermatohistologic diagnostics by Multi Epitope Ligand Cartography (MELC)
In-situ-topoproteome analysis of cutaneous lymphomas: Perspectives of assistance for dermatohistologic diagnostics by Multi Epitope Ligand Cartography (MELC)
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DOI：
10.1111/j.1610-0387.2007.06754.x
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发表时间：
2008-12-01
影响因子：
3.6
通讯作者：
Gollnick, Harald
中科院分区：
文献类型：
--
作者：
Bonnekoh, Bernd;Pommer, Ansgar J.;Gollnick, Harald
背景：免疫分型对皮肤淋巴瘤的诊断至关重要。为此，我们提出了一种适合皮肤组织的MELC技术应用平台。患者和方法：这种拓扑蛋白质组分析允许至少n = 100个表位原位亚细胞共定位。为此，将标本用Toponome成像循环器(R)处理，重复以下循环n次：1)用荧光团标记的抗体染色，2)荧光成像，3)光漂白。荧光图像的叠加和二值化导致组合分子表型（CMP），这涉及到一个像素或微地形单位（450 x 450 nm(2)， 20倍物镜）。皮肤活检来自蕈样真菌病（斑块/斑块病变）、牛皮癣、特应性湿疹患者和健康皮肤供者。结果：面向who - eortc皮肤淋巴瘤分类，建立了包含23个标记物的melc文库。根据一个开创性的详细程序，CMP频率在100 μ m水平蒙皮宽度的归一化中确定。通过TopoMiner策略，蕈样真菌病可以从其他状态中分离出来，具有最大的意义(p
Background: Immunophenotyping is essential for diagnostics of cutaneous lymphomas. In this regard we present a skin tissue-adapted application platform of MELC technology.Patients and Methods: This topoproteome analysis allows the subcellular colocalization of at least n = 100 epitopes in situ. For this purpose the specimen is processed by a Toponome Imaging Cycler (R) for a n-fold repetition of the following cycle: 1) staining with a fluorophore-labeld antibody, 2) fluorescence-imaging, and 3) photobleaching. Overlay and binarization of fluorescence images lead to combinatorial molecular phenotypes (CMP), which relate to a pixel or microtopographic unit (450 x 450 nm(2), 20x objective). Skin biopsies were derived from patients with mycosis fungoides (patch/plaque lesions), psoriasis, atopic eczema and from healthy skin donors.Results: In orientation to the WHO-EORTC-classification of cutaneous lymphomas a MELC-library of 23 markers was established. According to an inaugurative detailed procedure the CMP frequency was determined in a normalization to 100 mu m horizontal skin width. By a TopoMiner strategy mycosis fungoides could be separated from the other states with a maximum of significance (p