BIOSAFETY

The use of analytic flow cytometers and, especially, aerosol-generating sorters with samples that contain viable infectious organisms presents a serious infectious risk to the Facility users and staff. With the analytic flow cytometers the main problem is with contamination of external surfaces or a sample spill onto a work surface or floor in an environment not set up to handle this. This is especially true in our Facility where there is high traffic around the instruments. With the sorters it is this, as well as, the possibility for tremendous uncontrolled aerosol generation.

We require a copy of your SCHEDULE F for any biohazardous materials brought into the core.

ANALYZERS

The use of the  LSRII and the CyAns with un-fixed biohazardous samples is strictly prohibited. All such samples must be sterilized with the use of an approved agent - usually 1-2% formaldehyde. Examples of materials that are not to be used in a viable state on these instruments are human cells of any type (includes human cell lines as well as primary cells) whether it is known or unknown if the cells contain infectious agents (e,g, HIV, SIV, TB, HepB, HepC, HepD, LCMV, SARS, HTLV-a, HTLV-2) and cells with recombinant DNA vectors (e.g. lentivirus, retrovirus). If you are in doubt please discuss this with the Facility Director or contact EHS.

FACS Calibur in 613 and Stratedigm in 610D MEJB: Samples containing viable BSL-2 agents (or samples that could contain such agents but whose status is unknown) may be run on the FACS Calibur or Stratedigm. Prior to running these samples the investigator must complete the "Biosafety Form", discuss the experiments with the Facility Director, and adhere to the rules of using the flow cytometer with these samples. The rules are:

  • PPE - latex/nitrile gloves, lab coat and closed toe shoes are to be used at all times.
  • Transporting samples Samples are to be transported to and from the FACScan room in an approved biosafety transport container.
  • Spills If any sample is spilled in the room it is to be reported to Facility staff and cleaned up using bleach provided in a clean up kit.
  • Disposal No samples or other contaminated materials are to be disposed of down the sink.

                Any contaminated materials should be discarded into the red biosafety bag provided.

                Left over samples are to be returned to the user's lab using the approved transport container.

  • Disinfection When finished, the FACScan sample line is to be disinfected with bleach for 5 minutes minimum - use the high sample rate setting.

SORTERS 

The use of biohazardous samples on the sorters is much more of an issue than with the closed system analyzers. In sorting we are intentionally generating an aerosol. Under most conditions the aerosol is more or less controlled but some is always generated that drifts away from the instrument. When the stream hits a solid surface - as may happen when there is a partial clog in the nozzle tip - the amount of aerosol that can be generated is tremendous.

The sorting of viable biohazardous materials (including any primary human cells) may NOT be performed on either the MoFlo or the MoFlo XDP.

BSL-2 designated samples may only be sorted on the iCyt Reflection or FACS Aria.

To schedule a sort requires requires completion of a "Sorting Request Form" including disclosure of any potential biohazards ("Biosafety Form"). If you have any questions about your sorting of biohazardous samples please contact the Facility Director (nancy_fisher@med.unc.edu; 6-1530).