The University of Iowa Carver College of Medicine
Sorting or Analyzing Potentially
The following outlines the policy for sorting or analyzing live primary human cells, potentially infectious agents, cells infected with replication incompetent virus, or cells from animals previously exposed to infectious agent using the FACS DiVa or FACS Aria.
Flow cytometry instruments that use jet-in-air fluidics produce aerosols, especially when sorting. These aerosols may contain infectious agents, such as hepatitis or HIV, when sourced from live human tissue such as PBL's or bone marrow. In addition, sorting known infectious agents such as attenuated tuberculosis bacteria or cells transfected or transformed with infectious agents such as Epstein-Barr Virus also pose a risk from aerosol exposure. Since this aerosol is under pressure, potential for infectivity is theoretically much higher than incidental exposure. In order to protect our instrument operators when sorting or analyzing potentially infectious samples, we are asking all labs that use the Facility to follow these guidelines.
- Prior approval for using a cell sorter (FACS DiVa, FACS Aria II) to sort or analyze live primary human samples, cells from animals previously exposed to an infectious agent, or cultured cells exposed to potentially infectious agents should be obtained from the director of the facility, Dr. Zuhair Ballas (firstname.lastname@example.org). Cells transformed with BSL-2 replication incompetent virus do not need pre-approval before sorting.
- The Environmental Health and Safety Office (EHS) is requiring the Facility to obtain a copy of the primary investigator's approved rDNA registration documentation before sorting cell samples transfected with rDNA. The documentation should include a list of vectors showing the biosafety level of each. The purpose of this documentation is to help the Facility determine the biosafety level of the cells. The cell sorter can then be properly configured for biohazard containment and appropriate personal protection equipment worn by Facility personnel. Please provide the requested documentation to the Facility Technical Director, Justin Fishbaugh, in electronic format (PDF) by email to email@example.com before coming to the Facility for the cell sorting appointment.
- All live primary human samples will be sorted using the Becton Dickinson FACS Aria II (not the FACS DiVa) which utilizes a biosafety cabinet to isolate the instrument operator from potential exposure to aerosols.
- The Facility cannot sort samples that have been exposed to BLS-3 agents or are considered BSL-3.
- A Risk Factor form available from the Flow Cytometry Facility website will be filled out by the principal investigator for each new patient, potentially infectious agent, cells infected with replication incompetent virus, or cells from animals previously exposed to an infectious agent and returned to the facility before the sort (or analysis using one of the cell sorters). This includes "normal" control subjects. A separate form must be filled out for each sort.
- When sorting live primary human samples, a diagnosis for the person from whom the sample was obtained must be included on the risk factor form. This will help the Facility assess the biosafety level of the samples. The Facility cannot sort samples from patients known to be infected with the following infectious agents or any other agent that is considered BSL-3.
- HTLV I/II
- When scheduling cell sorter time (sorting or analyis) for live human samples, potentially infectious agents, cells infected with replication incompetent virus, or cells from animals previously exposed to an infectious agent, post sort decontamination time will be added to the schedule automatically by the online reservation system. Charges for this time will be assessed at the regular charge rate.
- Live human samples, potentially infectious agents, cells infected with replication incompetent virus, or cells from animals previously exposed to an infectious agent that are prepared for analysis only, will be run on the BD FACScan or LSR II which do not produce aerosols. The Facility must be informed that these samples will be run live via the online reservation system, but will NOT require infectious disease testing or completion of a Risk Factor form. For fluorochrome combinations that require analysis, but not sorting, using the Becton Dickinson FACS DiVa or FACS Aria (cell sorters), all of the above guidelines must be followed.
Note: The above policy does not apply to well characterized cell lines (e.g. those obtained from ATCC). The policy does apply to new cell lines derived by transformation with agents such as Epstein-Barr Virus and not subjected to rigorous genetic analysis.