van den Engh G, Stokdijk W
Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore, California 94550.
Cytometry. 1989 May;10(3):282-93. doi: 10.1002/cyto.990100307.
This report describes the data acquisition electronics for a flow cytometer. The design differs from most instruments in that the signals from a large number of detectors are processed in parallel. Each of the input channels is capable of autonomously measuring and digitizing the fluorescence signals. The digitized values that belong to one particle are collected by digital circuitry and are presented as a compact data package on a special bus. In addition to the pulse values, the data package contains a time marker, information needed for sort decisions, and an error detection code. Specially designed electronic modules that read the information from the bus can take complex multiparameter sort decisions at a very high speed. All events can also be recorded as data lists by a computer. The lists can be used to reconstruct a sort or analysis run. The raw data lists can also be reduced to kinetic curves and/or (gated) multivariate histograms. As a result of the applied scheme of parallel pulse processing, the dead time of the system is independent of the number of parameters measured and the number and time separation of the excitation beams. The instrument has a cycle time of 5 microseconds, which corresponds to a throughput rate of 2 x 10(5) events/s. At this rate, the incidence of correlation errors is well below 1 in 10(8) analyzed particles. The system has proved to be reliable and convenient to use in a variety of experiments. Its high speed and low error rate make it well suited for high-resolution measurements, rare-event analysis, kinetic measurements, and high-speed cell sorting.
本报告描述了一种流式细胞仪的数据采集电子设备。该设计与大多数仪器的不同之处在于,来自大量探测器的信号是并行处理的。每个输入通道都能够自主测量荧光信号并将其数字化。属于一个粒子的数字化值由数字电路收集,并作为一个紧凑的数据包在一条特殊总线上呈现。除了脉冲值外,数据包还包含一个时间标记、分选决策所需的信息以及一个错误检测代码。专门设计的从总线上读取信息的电子模块能够以非常高的速度做出复杂的多参数分选决策。所有事件也可以由计算机记录为数据列表。这些列表可用于重建分选或分析过程。原始数据列表还可以简化为动力学曲线和/或(选通的)多变量直方图。由于采用了并行脉冲处理方案,系统的死时间与测量的参数数量以及激发光束的数量和时间间隔无关。该仪器的周期时间为5微秒,对应于2×10⁵个事件/秒的通量率。以这个速率,相关误差的发生率在分析的粒子中远低于1/10⁸。该系统已被证明在各种实验中可靠且使用方便。其高速度和低错误率使其非常适合高分辨率测量、稀有事件分析、动力学测量和高速细胞分选。