Automated image cytometry for detection of rare, viral antigen-positive cells in peripheral blood.
Automated image cytometry for detection of rare, viral antigen-positive cells in peripheral blood.
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DOI：
10.1002/cyto.990150304
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发表时间：
1994-03
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影响因子：
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通讯作者：
J. Ploem‐Zaaijer;W. Mesker;G. Boland;W. Sloos;F. M. van de Rijke;M. Jiwa;A. Raap
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文献类型：
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作者：
J. Ploem‐Zaaijer;W. Mesker;G. Boland;W. Sloos;F. M. van de Rijke;M. Jiwa;A. Raap
描述了一种基于自动筛选的细胞检测方法，用于识别人类外周血中免疫酶标记的白细胞的低频（10万分之一）。所使用的图像细胞仪（LEYTAS）包括一台广角全自动显微镜（Autoplan）和一台模块化图像分析计算机（MIAC），均来自德国Wetzlar的徕卡公司。MIAC包含最佳使用数学形态学算法的图像板。与MIAC的通信是通过个人电脑进行的。用C语言编写了自动细胞分析程序。该系统的主要特点是对大型显微镜视场的快速分析，包括所有细胞的计数，感兴趣对象的选择（报警），以及显示这些报警的数字存储图像。我们在50例肾移植受者血液涂片中检测了表达巨细胞病毒抗原（pp65）的白细胞。DAB免疫酶（过氧化物酶）染色，苏木精反染。为了测定灵敏度，进行了一系列阳性样品与阴性样品的稀释。最低检出率为1个抗原阳性细胞/3 × 10(5)个抗原阴性细胞。筛选时间约60 min， 100万个细胞。
A cell detection method based upon automated screening is described for recognition of low frequencies (1 in 100,000) of immuno-enzymatically labelled white blood cells in human peripheral blood. The used image cytometry instrumentation (LEYTAS) includes a wide-field, fully automated microscope (Autoplan) and a modular image analysis computer (MIAC), both from Leica, Wetzlar, Germany. The MIAC contains image boards for optimum use of mathematical morphology algorithms. Communication with the MIAC is via a personal computer. Programs for automated cell analysis have been written in C language. Main features of the system are fast analysis of large microscope fields including a count of all cells, selection of objects of interest (alarms), and display of digitally stored images of these alarms. We tested this system for the detection of white blood cells expressing antigen of cytomegalovirus (pp65) in 50 human blood smears from kidney transplant recipients. Immuno-enzymatic (peroxidase) staining was performed with DAB and counterstaining with hematoxylin. For determination of the sensitivity, a series of dilutions of a positive sample with a negative sample was performed. The lowest frequency detected was 1 antigen-positive cell/3 x 10(5) antigen-negative cells. Screening time was about 60 min for one million cells.