International Society for the Advancement of Cytometry cell sorter biosafety standards

Abstract

Flow cytometric cell sorting of biological specimens has become prevalent in basic and clinical research laboratories. These specimens may contain known or unknown infectious agents, necessitating precautions to protect instrument operators and the environment from biohazards arising from the use of sorters. To this end the International Society of Analytical Cytology (ISAC) was proactive in establishing biosafety guidelines in 1997 (Schmid et al., Cytometry 1997;28:99–117) and subsequently published revised biosafety standards for cell sorting of unfixed samples in 2007 (Schmid et al., Cytometry Part A J Int Soc Anal Cytol 2007;71A:414–437). Since their publication, these documents have become recognized worldwide as the standard of practice and safety precautions for laboratories performing cell sorting experiments. However, the field of cytometry has progressed since 2007, and the document requires an update. The new Standards provides guidance: (1) for laboratory design for cell sorter laboratories; (2) for the creation of laboratory or instrument specific Standard Operating Procedures (SOP); and (3) on procedures for the safe operation of cell sorters, including personal protective equipment (PPE) and validation of aerosol containment.

A method permissive to fixation and permeabilization for the multiparametric analysis of apoptotic and necrotic cell phenotype by flow cytometry

“…The asymmetry was shown to increase the fluorescence of merocyanine 540, a negatively charged chromophore that is thought to interact with the outer leaflet of the plasma membrane (10) and PS exposure has been used to detect apoptotic cells using PS specific ligands such as the biotinylated C2A domain of synaptotagmin I (11), fluorescent Zn 21 -dipicolylamines (12,13) or fluorescent Annexin-V. Third, the test cannot be safely performed in cells infected in vitro or in vivo with pathogens that could be aerosolized during flow cytometry analysis or cell sorting (19,20). This method offers the great advantage to discriminate early apoptotic cells, revealed as Annexin-V positive only, from late apoptotic cells, revealed as double Annexin-V and PI staining and single PI positive necrotic cells, and to simultaneously allow membrane phenotype characterization using additional fluorochrome matched monoclonal antibodies.…”

A simple and biosafe method for isolation of human umbilical vein endothelial cells

“…It is widely recognized that clinical samples may contain known or unknown infectious agents [12].To protect the technicians, in 2007 the International Society of Analytical Cytology published revised biosafety standards for cell sorting of unfixed samples [13]. They recommend performing a risk assessment before starting an experiment and using personal protective equipment during the process.…”

Analyzing Persister Physiology with Fluorescence-Activated Cell Sorting

“…Additionally, consideration must be given to the risk factor group and biosafety level designation of the organisms to be sorted. Cell sorting creates aerosols through droplet formation causing the potential risk for inhalation exposure, and the system can be under high pressure increasing the risk of splash exposure to liquids (27–29). Biosafety professionals should be consulted and proper precautions should be in place prior to conducting any FACS experiments.…”