Sheath Contamination Survey: An Examination of Common (Though Not Necessarily Best) Laboratory Practices A Study of the Flow Cytometry Research Group (FCRG) Dave Adams, University of Michigan; Alan Bergeron, Dartmouth College; Laura Blunk, Stowers Institute for Medical Research; Kathleen Brundage, West Virginia University; Karen Clise-Dwyer, MD Anderson Cancer Center; Matt Cochran, University of Rochester Medical Center; Monica De. Lay, Cincinnati Children’s Hospital; Roxana Del Rio Guerra, University of Vermont; Maris Handley, Massachusetts General Hospital; Peter Lopez, New York University Langone Medical Center; E. Michael Meyer, University of Pittsburgh Cancer Institute; Alan Saluk, The Scripps Research Institute EB: Frances Weis-Garcia
Previous FCRG Study looking at common sorter contaminants • Develop effective endotoxin cleaning procedure • Test for additional contaminants • Poll SRLs beyond the FCRG and evaluate common practices
The Callout • Survey Monkey Platform • Advertised on PUCL mailing list and Google+ Cytometry Community – Bribed with chance to win Starbucks! • Participants asked about their testing procedures, operating procedures and maintenance procedures • Participants asked to submit samples for testing
How’d We Do? • 61 Respondants – 3 from Non-SRLs • 106 Sorters Described • Several Lessons About How Not to Ask a Question in a Survey • 3 Happy Coffee Drinkers
Instrumentation Quick Facts • Of the 106 cell sorters described in the survey: • 54 are housed within Biosafety Cabinets • 55 are operated solely by laboratory staff while the remaining 51 allow trained users to operate the instrument • Average reported instrument usage is 76 -100% capacity though that number falls to 51 -75% as the number of sorters within the facility increases. • Sheath Sources break down as follows: • 51% of respondents use a commercial 1 x saline product • 35% use a commercial concentrate saline product which is then diluted in-house • 14% manufacture their saline in-house
Testing Frequency Daily Weekly Monthly Never Only Upon Reported Contamination Responses Percentage 2 11 7 8 25 4 21 13 15 47
Testing Location Collection Location Sheath Tank Flow Cell Sample Introduction Area Bulk Sheath container Responses 24 40 15 Percentage 53 89 33 9 20
Self-Reported Positive Contamination Results in Previous Twelve Month Period 25 21 20 Responses 17 15 10 5 3 3 2 1 0 I do not test 0 1 -2 3 -5 5 -10 10+
User Operation
Bacteria Testing
Negative + ++ +++ Sheath Tank 2 6 4 5 Stream 12 2 0 3
RNase Testing
Where to From Here? 1. Nearly half the survey respondents do not perform any proactive sheath testing and yet over half the received samples tested positive for bacterial/fungal contamination. 2. The prevalence of RNase in our testing pool was low, but it is worth noting that both positive tests were recorded in facilities that do no RNase testing. 3. Survey results at this point are limited and additional responses will present a better idea of current standard practices.
Acknowledgements We would like to thank all the individuals who answered our survey and provided samples for our study. Follow the QR code to The current survey