Clatch R J, Foreman J R
Christ Hospital and Medical Center, Oak Lawn, Illinois, USA.
Cytometry. 1998 Feb 15;34(1):36-8.
Laser scanning cytometry is a new laboratory technology similar to flow cytometry but with advantages for certain clinical and research applications. To date, laser scanning cytometry has been successfully used to perform three-color immunophenotypic analysis of hematologic specimens, single-color immunophenotyping plus DNA content analysis of numerous specimen types, and automated analysis of fluorescence in situ hybridization specimens. Several other interesting applications are also in development. In general, advantages of laser scanning cytometry include reduced specimen size requirements, simplified methodologies, and the ability to microscopically examine individual cells-allowing for the direct correlation of cytologic morphology with objective fluorescence measurements. In this report, we describe a method which more fully takes advantage of the laser scanning cytometer's capabilities for immunophenotypic analysis of hematologic specimens. Specifically, we have devised a method to increase the number of fluorescent parameters from three to a total of six, five representing binding of immunofluorescent antibodies and one for stoichiometric measurements of DNA content. As with most laser scanning cytometric applications, this technique can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology.
激光扫描细胞术是一项新的实验室技术，与流式细胞术类似，但在某些临床和研究应用方面具有优势。迄今为止，激光扫描细胞术已成功用于对血液学标本进行三色免疫表型分析、对多种标本类型进行单色免疫表型分析及DNA含量分析，以及对荧光原位杂交标本进行自动化分析。其他一些有趣的应用也在开发中。一般来说，激光扫描细胞术的优势包括对标本大小要求降低、方法简化，以及能够在显微镜下检查单个细胞——从而使细胞学形态与客观荧光测量直接相关。在本报告中，我们描述了一种能更充分利用激光扫描细胞仪对血液学标本进行免疫表型分析能力的方法。具体而言，我们设计了一种方法，将荧光参数的数量从三个增加到总共六个，其中五个代表免疫荧光抗体的结合，一个用于DNA含量的化学计量测量。与大多数激光扫描细胞术应用一样，该技术可用于极小的标本，并能使所有测量的荧光参数与光学显微镜下的细胞学形态直接相关。
