Department of Immunology and Immunogenetics, Southmead Hospital, Westbury-on-Trym, Bristol BS10 5NB.
Ann Clin Biochem. 2012 Jan;49(Pt 1):17-28. doi: 10.1258/acb.2011.011128. Epub 2011 Oct 25.
Flow cytometry has had an impact upon all areas of clinical pathology and now, in the 21st century, it is truly coming of age. This study reviews the application of flow cytometry within clinical pathology with an emphasis upon haematology and immunology. The basic principles of flow cytometry are discussed, including the principles and considerations of the flow-cell and hydrodynamic focusing, detector layout and function, use of fluorochromes and multicolour flow cytometry (spectral overlap and colour compensation), alongside the strategies available for sample preparation, data acquisition and analysis, reporting of results, internal quality control, external quality assessment and flow sorting. The practice of flow cytometry is discussed, including the principles and pitfalls associated with leukocyte immunophenotyping for leukaemia and lymphoma diagnosis, immune deficiency, predicting and monitoring response to monoclonal antibody therapy, rare event detection and screening for genetic disease. Each section is illustrated with a case study. Future directions are also discussed.
流式细胞术已经对临床病理学的各个领域产生了影响,现在,在 21 世纪,它真正地成熟了。本研究回顾了流式细胞术在临床病理学中的应用,重点是血液学和免疫学。讨论了流式细胞术的基本原理,包括流室和流体力学聚焦、探测器布局和功能、荧光染料的使用和多色流式细胞术(光谱重叠和颜色补偿)的原理和注意事项,以及用于样品制备、数据采集和分析、结果报告、内部质量控制、外部质量评估和流式分选的策略。讨论了流式细胞术的实践,包括与白血病和淋巴瘤诊断、免疫缺陷、预测和监测单克隆抗体治疗反应、稀有事件检测和遗传疾病筛查相关的白细胞免疫表型分析的原理和陷阱。每个部分都用案例研究来说明。还讨论了未来的方向。