Metabolism Unit and KI/AZ Integrated Cardio Metabolic Center, Department of Medicine, Karolinska University Hospital, Karolinska Institutet, Stockholm, SE 171 76, Sweden.
Flow Cytometry Core Facility, Van Andel Research Institute, Grand Rapids, Michigan, 49503, USA.
Cytometry A. 2020 Aug;97(8):800-810. doi: 10.1002/cyto.a.23998. Epub 2020 Mar 9.
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.
细胞仪的特性分析对于定义数据生成的可靠和可重现的操作范围至关重要。现有的仪器优化和特性分析协议是为依赖光电倍增管 (PMTs) 进行光子检测的细胞仪开发的。最近，仪器制造商已开始在仪器中采用雪崩光电二极管 (APD) 替代 PMTs。两种探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 特性分析协议不适用于基于 APD 的仪器。在本文中，我们测试了（在两个不同地点的三台机器）各种方法，以确定优化 Beckman Coulter CytoFLEX™（CytoFLEX）上 APD 的最佳方法。在此基础上，我们提出了易于实施的 CytoFLEX 特性分析和操作指南。这些协议不是为了比较基于 APD 和 PMT 的系统，也不是为了直接比较不同的 CytoFlex 仪器。遵循这些协议将允许 CytoFLEX 用户对其仪器进行特性分析，并帮助确定优化设置，以生成一致和可重复的数据。
