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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅴ, Jan 23-25, 2002, San Jose, USA >Novel approaches for immunophenotyping by laser scanning cytometry (LSC)
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Novel approaches for immunophenotyping by laser scanning cytometry (LSC)

机译：激光扫描细胞仪（LSC）进行免疫表型鉴定的新方法

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摘要

LSC is a microscope-based technology. The principle of the instrument is that any specimen is immobilized on a microscope slide. Therefor the cells are not lost in a fluid stream but are kept on the slide and minimal specimens as low as 1.000 cells can be analyzed. Additionally cells are available for further analyses such as staining for another set of specific markers and re-analysis or cytological staining (H&E). This approach multiplies the information gained from a given sample. We have established an assay for immunophenotyping of peripheral blood leukocytes by LSC. Cells are prepared according to routine flow cytometry protocols with a first set of CD-antibodies and are fixed on microscope slides. As a stable trigger signal the nuclear DNA is stained by 7-aminoactinomycin-D. This guarantees that all nucleated cells and that only nucleated cells are included in the analysis, and may differentiate between lymphocytes and neutrophiles by staining intensity. After analysis cells are stained with a second set of CD-antibodies and analyzed again. This step can be repeated with a third set of CD-antigens. Since the location of the cells on the slide is fixed data from the analyses can be attributed to the same cell.
机译：LSC是基于显微镜的技术。该仪器的原理是将任何样本固定在显微镜载玻片上。因此，细胞不会在流体流中丢失，而是保留在载玻片上，并且可以分析低至1.000个细胞的最小样本。另外，细胞可用于进一步分析，例如另一组特异性标志物的染色以及重新分析或细胞学染色（H＆E）。这种方法将从给定样本中获得的信息相乘。我们建立了一种通过LSC对外周血白细胞进行免疫表型分析的方法。根据常规流式细胞术方案，用第一组CD抗体制备细胞，并将其固定在显微镜载玻片上。作为稳定的触发信号，核DNA被7-氨基放线菌素D染色。这保证了所有有核细胞并且仅有核细胞都包括在分析中，并且可以通过染色强度区分淋巴细胞和嗜中性粒细胞。分析后，将细胞用第二组CD抗体染色并再次分析。可以使用第三组CD抗原重复此步骤。由于单元格在幻灯片上的位置是固定的，因此来自分析的数据可以归因于同一单元格。

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