Cytometer characterization is critical to define operational bounds within which the data generated are reliable and reproducible. Existing instrument optimization and characterization protocols were developed for cytometers relying on photomultiplier tubes (PMTs) for photon detection. Recently, instrument manufacturers have begun incorporating avalanche photodiodes (APDs) in place of PMTs. Differences in noise and signal amplification properties of the two detector types make many of the established PMT characterization protocols inappropriate for APD‐based instruments. In this article, we tested (three machines on two different sites) a variety of approaches to determine the best method for APD optimization on the Beckman Coulter CytoFLEX™ (CytoFLEX). From this, we propose easy‐to‐implement guidelines for CytoFLEX characterization and operation. These protocols are not designed to compare APD versus PMT based systems, nor are they designed to directly compare different CytoFlex instruments. Following these protocols will allow CytoFLEX users to characterize their instruments and help to identify optimized settings that allow for the generation of consistent and reproducible data. © 2020 International Society for Advancement of Cytometry

中文翻译：

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Beckman Coulter CytoFLEX™ 平台优化和表征的实用指南。

细胞仪表征对于定义操作范围至关重要，在该范围内生成的数据是可靠且可重复的。现有的仪器优化和表征协议是为依赖光电倍增管 (PMT) 进行光子检测的细胞仪开发的。最近，仪器制造商已开始采用雪崩光电二极管 (APD) 来代替 PMT。两种检测器类型的噪声和信号放大特性的差异使得许多已建立的 PMT 表征协议不适用于基于 APD 的仪器。在本文中，我们测试了（在两个不同站点上的三台机器）各种方法，以确定在 Beckman Coulter CytoFLEX™ (CytoFLEX) 上进行 APD 优化的最佳方法。由此，我们提出了易于实施的 CytoFLEX 表征​​和操作指南。这些协议并非旨在比较基于 APD 与 PMT 的系统，也并非旨在直接比较不同的 CytoFlex 仪器。遵循这些协议将允许 CytoFLEX 用户表征他们的仪器并帮助确定允许生成一致和可重复数据的优化设置。© 2020 国际细胞计量学促进会