MICROSCOPE-BASED MULTIPARAMETER LASER SCANNING CYTOMETER YIELDING DATA COMPARABLE TO FLOW-CYTOMETRY DATA
MICROSCOPE-BASED MULTIPARAMETER LASER SCANNING CYTOMETER YIELDING DATA COMPARABLE TO FLOW-CYTOMETRY DATA
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DOI：
10.1002/cyto.990120502
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发表时间：
1991-01-01
期刊：
影响因子：
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通讯作者：
KAMENTSKY, LD
中科院分区：
文献类型：
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作者：
KAMENTSKY, LA;KAMENTSKY, LD
我们描述了一种计算机控制的10 μ m光斑大小的激光扫描细胞仪，用于对表面上的无约束细胞（如显微镜载玻片）进行多波长荧光和散射测量。放置在显微镜载物台上的载玻片的指定区域被自动扫描，并且产生高于阈值的散射或荧光值的细胞被发现并被单独处理以确定测量参数的列表。对于每个荧光或散射测量参数，此列表包含以细胞为中心的扫描窗口的积分值和峰值以及位模式图像。测量时间、细胞在载玻片上的位置和两个分割指数也包含在列表中。测量时间、单元位置和从位模式导出的属性可与积分或峰值测量值互换使用，作为多属性显示的坐标。可以选择细胞进行计数，以各种形式显示数据，或基于它们在两个属性屏幕的链中满足复杂标准的视觉观察。可以在实验期间或回顾性地观察具有选定特性的细胞。指定的样本区域可重复重新测量，以进行动力学细胞研究。已经构建了氩离子和基于HeNe的激光仪器，并且已经编写和评估了软件，其具体目标是提高碘化丙啶染色的细胞DNA测量的精度。描述了该仪器的一些功能及其当前性能。
We describe a computer-controlled 10-mu-m spot size laser scanning cytometer for making multiple wavelength fluorescence and scatter measurements of unconstrained cells on a surface such as a microscope slide. Designated areas of slides placed on a microscope stage are automatically scanned, and cells which generate above-threshold scatter or fluorescence values are found and individually processed to determine a list of measurement parameters. For each fluorescence or scatter measurement parameter, this list contains the integrated and peak values and bit pattern images of a scan window centered on the cell. The measurement time, the position of the cell on the slide, and two segmentation indices are also included in the list. Measurement time, cell position, and properties derived from the bit patterns are used interchangeably with integrated or peak measurement values as coordinates of multiproperty displays. Cells may be selected for counting, data display in various forms, or visual observation based on their meeting complex criteria among a chain of two property screens. Cells with selected properties may be viewed during an experiment or retrospectively. A designated specimen field may be repeatedly remeasured to perform kinetic cell studies. An argon ion and a HeNe- based laser instrument have been constructed and software has been written and evaluated with the specific goal of increasing the precision of propidium iodide-stained cellular DNA measurements. Some of the capabilities of the instrument and its current performance are described.