Verifying Proper Function of the Aerosol Evacuation System Prior to Sorting Potentially Infectious Samples
Kristen M. Reifel, Avrill Aspland, Suat Dervish, Iyadh Douagi, Alyssa C. Fears, Evan R. Jellison, Cecily C. Midkiff, Taryn Mockus-Daehn, Matilda J. Moström, Michael Solga, Brandon K. Swan, James Thomas, Stephen Perfetto
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Abstract
High concentrations of aerosols can be generated within the sort collection area of cell sorters during instrument failures that cause the stream to deviate, such as a partial nozzle obstruction. Complete containment of these aerosol particles becomes essential for operator safety when working with potentially infectious or hazardous samples. Currently, aerosol containment is accomplished through the generation of continuous negative airflow within the sort collection area using an aerosol evacuation system, which can be enhanced by using primary containment devices such as biosafety cabinets. Unlike biosafety cabinets, many aerosol evacuation systems are not certified or tested on a regular basis after installation. Therefore, proper function of the system must be verified by the user prior to running hazardous samples to ensure that it is operational and provides sufficient protection for the operator. This protocol describes an updated procedure for verifying the containment and evacuation of aerosols generated when the stream is disrupted during an instrument failure. In this procedure, aerosols are generated to simulate a partial nozzle obstruction while running 1-µm fluorescent beads. Air samples are collected just outside the sort collection area using a disposable impactor-style aerosol sampler cassette and are examined for the presence of beads in an effort to detect aerosols. If no beads are present, aerosols were adequately contained and evacuated by the aerosol evacuation system. The presence of beads, however, indicates a potential failure of the aerosol evacuation system and/or other engineering controls that could result in the exposure of laboratory workers to any infectious or hazardous samples that are run through the instrument. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.
Basic Protocol : Validation of the aerosol evacuation system using 1-µm fluorescent beads and disposable aerosol sampler cassettes
Support Protocol : Preparation of 1-µm fluorescent bead reference slide
在分拣具有潜在传染性的样品之前,验证气溶胶疏散系统的正确功能。
在仪器故障(如部分喷嘴阻塞)导致气流偏离时,可以在细胞分选器的分类收集区域内产生高浓度的气溶胶。在处理潜在传染性或危险样品时,完全控制这些气溶胶颗粒对操作人员的安全至关重要。目前,气溶胶密封是通过使用气溶胶疏散系统在分类收集区域内产生连续的负气流来完成的,可以通过使用生物安全柜等主要密封装置来加强。与生物安全柜不同,许多气溶胶疏散系统在安装后没有经过认证或定期测试。因此,在运行危险样品之前,必须由用户验证系统的正确功能,以确保它是可操作的,并为操作人员提供足够的保护。本协议描述了一个更新的程序,用于验证在仪器故障期间流中断时产生的气溶胶的遏制和疏散。在此过程中,产生气溶胶以模拟在运行1µm荧光珠时喷嘴的部分阻塞。使用一次性冲击式气溶胶采样器盒在分类收集区域外收集空气样本,并检查是否存在珠子以检测气溶胶。如果没有珠子存在,则气溶胶被气溶胶疏散系统充分地包含和疏散。然而,珠子的存在表明气溶胶疏散系统和/或其他工程控制的潜在故障,可能导致实验室工作人员暴露于通过仪器运行的任何传染性或危险样品。©2025作者。当前协议由Wiley期刊有限责任公司出版。本文由美国政府雇员贡献,他们的工作在美国属于公有领域。基本方案:使用1µm荧光珠和一次性气溶胶取样盒验证气溶胶抽吸系统。支持方案:制备1µm荧光珠参比载玻片。
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