Biosafety and User Guidelines

Rangos Flow Cytometry BSL2+ Biosafety and User Guidelines

  • Personal Protective Equipment
  1. Strict enforcement for the use of lab coats and gloves
  2. Additional signage when BSL2+ specimens are analyzed/sorted
  3. As needed, N-9 mask or Safety respiratory may worn by core staff or self-user of the sorters during live cell sorting of certain BSL2+ specimens
  • Regardless of background/project, first-time users must arrange a consultation meeting.
  • All users must complete a job request form including account number information.
  • Scheduling is required for all services. Plan experiments appropriately.
  1. No-shows are charged for the full time reserved.
  2. An overtime charge for a full hour is levied when cytometry work spills over into the next pre-scheduled time slot. Any unfinished cytometry may re-commence at next available time slot.
  3. Unused time of a reserved block in excess of full hour will incur a charge.
  4. A lab may schedule only up to 3-half day blocks of sorting or analysis time for any given week. Excess reservation is subject to automatic cancellation during high-demand. Users may contact the Core Staff to check for extra scheduling.
  • Self-users (Fortessa and LSRII): Training is required/provided by the Core Staff.
  • Cell sorting during business hours is staff-assisted. Off-hours sorting may be done with prior arrangement.
  • “Super-user” designation is given to those who complete more rigorous training on the cell sorters. A super-user has privilege for cell sorting during off-hours.
  • All specimens for analysis must be fixed with at least 2% paraformaldehyde (in a suitable buffer) for at least 20 minutes. If cell analysis requires live cells, then the analysis must be moved to the cell sorters.
  • All live-cell sorting of BSL2+ specimens must be disclosed.
  • To minimize clogging of instruments, users must filter their specimens using a suitable nylon mesh filter (contact the Core Staff for information).
  • Regardless of BSL classification, specimens containing free viral particles in solution will not be accepted. Users employing viruses or viral vectors MUST certify the type of viral/vector used and the absence of viral particles in specimens.
  • Live cell sorting of BSL1 or 2 or 2+, virus-/bacteria-infected specimens (http://www.ibc.pitt.edu/home/viral-standards) require prior arrangement/notification about the type of infectious agent.
  • Users must collect any residual specimens after completion of cytometry work.