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Biohazards in a flow cytometry facility can arise either from sample handling or from the use of the flow cytometer. Although most samples that are analyzed in the flow cytometry laboratory are fixed, some protocols require that cells are run through the instruments unfixed, e.g.
- certain apoptosis assays
- measurement of calcium flux
- viable cell sorting
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Specimens can contain either known or unknown pathogens. Most of the infectious agents encountered in the specimens that are analyzed on analytic and sorting flow cytometers are transmitted by percutaneous or mucous membrane exposure. However, some may also be transmitted by inhalation of aerosols generated by the flow cytometer, which cannot be contained during analysis or sorting. Preparations of potentially infectious materials (bacteria, viruses or fungi) must be fixed prior to analysis to reduce the risk of contamination. Examples include cells known to be producing infectious viruses such as EBV, TB or HIV.
FACSCalibur flow cytometers in use in the Flow Cytometry Unit have biosafety features for reduced risk of operator exposure to instrument-generated sample droplets and aerosols(e.g., enclosed flow cells, droplet containment modules). However, operators of these instruments need to take care to securely place each sample tube into the sample introduction port, otherwise it could be blown off once it is pressurized and splash sample onto the operator.
The waste needs to be collected into a waste container which contains fresh concentrated household bleach in sufficient quantity to achieve a final concentration of 10% when the flask is full.
Routine decontamination of instrument fluid lines with freshly diluted (7%-10%) bleach solution is standard laboratory practice.
For protection of core laboratory personnel and users of the facility from exposure to occupational hazards, it is imperative that all users of the flow cytometry core facility comply with University of Stellenbosch Safety Regulations as outlined in the Flow Cytometry Unit Laboratory Safety Manual and in the Laser Safety Manual.
There is no provision for the analysis of radio-labeled cells, or cells which have been physically treated with radioisotopes and which pose a potential radiation safety hazard.
Users should also be aware that many dyes used for staining in flow cytometric protocols are toxins, mutagens or carcinogens.
Please note in the online log if your sample contains a hazardous chemical. You must provide a Material Safety Data Sheet (MSDS) for any hazardous chemical and Standard Operating Procedure when using the hazardous material.
Users are required to fill out the Biosafety Questionnaire (please print it, fill it out completely, sign it and return it to Mandy Alblas).
For the official safety guidelines of the International Society of Analytic Cytology (ISAC) for cell sorting of potentially and known biohazardous samples, refer to
"Biosafety Guidelines for Sorting of Unfixed Cells," Schmid et al., Cytometry 28: 99-117, 1997.
For further information on occupational safety issues and safety issues related to flow cytometry, please refer to the Laser Safety Manual , Radiation - Laser Hazards (United States Occupational Health and Safety Administration) and General Laboratory Health and Safety (Center for Disease Control).
Information related to practical biosafety issues for HIV immunophenotyping such as survival and inactivation of HIV under laboratory conditions, sample shipping, fixation, sample disposal, laser safety, flow cytometer fluid lines disinfection, and post-exposure management is available in the Q&A document "Biosafety Concerns for Flow Cytometric HIV Immunophenotyping" taken from the Adult AIDS Clinical Trial Group Website (Adobe Acrobat required) and from the publication by Schmid et al., "Biosafety Consideration for Flow Cytometric Analysis of Human Immunodeficiency Virus Infected Samples," Cytometry (Communications in Clinical Cytometry) 38:195-200, 1999.
