Standard Safety Practices for Sorting of Unfixed Cells

Ingrid Schmid1, Claude Lambert2, David Ambrozak3, Stephen P. Perfetto3

1 David Geffen School of Medicine at UCLA, Los Angeles, California, 2 University Hospital St. Etienne, St. Etienne, 3 National Institute of Health, Bethesda, Maryland
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 3.6
DOI:  10.1002/0471142956.cy0306s39
Online Posting Date:  January, 2007
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Cell sorting of viable biological specimens has become widespread in laboratories involved in basic and clinical research. As these samples can contain infectious agents, precautions to protect instrument operators and the environment from hazards arising from the use of sorters are paramount. This unit presents a revised and updated version of the biosafety guidelines for sorting of unfixed cells established in 1977 by the International Society of Analytical Cytology (ISAC), whose recommendations have become recognized worldwide as the standard practices and safety precautions for laboratories performing viable cellā€sorting experiments. The unit contains background information on the biohazard potential of sorting and the hazard classification of infectious agents as well as recommendations on (1) sample handling, (2) operator training and personal protection, (3) laboratory design, (4) cell sorter setup, maintenance, and decontamination, and (5) testing the instrument for the efficiency of aerosol containment.

Keywords: flow cytometry; occupational health; biohazards; cell sorting; biosafety; aerosol containment

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Literature Cited
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Almeida, J.D., Kulatilake, A.E., Mackay, D.H., Shackman, R., Chisholm, G.D., MacGregor, A.B., O'Donoghue, E.P., and Waterson, A.P. 1971. Possible airborne spread of serum‐hepatitis virus within a haemodialysis unit. Lancet 2:849‐850.
   Aloisio, C.H. and Nicholson, J.K.A. 1990. Recovery of infectious human immunodeficiency virus from cells treated with 1% paraformaldehyde. J. Immunol. Methods 128:281‐285.
   Andersen, A.A. 1958. New sampler for the collection, sizing, and enumeration of viable airborne particles. J. Bacteriol. 76:471‐484.
   Centers for Disease Control and Prevention. 1999. Biosafety in Microbiological and Biomedical Laboratories, 4th ed., U.S. Government Printing Office, Washington.
   Clinical and Laboratory Standards Institute, formerly National Committee for Clinical Laboratory Standards (NCCLS). 2005. Protection of laboratory workers from instrument biohazards and infectious disease transmitted by blood, body fluids, and tissue. Approved guideline. Doc. M29‐A3 (ISBN1‐56238‐567‐4).
   Collins, C.H. and Kennedy, D.A. 1999. Laboratory‐Acquired Infections. 4th ed. Oxford, Butterworth‐Heinemann.
   Druce, J.D., Jardine, D., Locarnini, S.A., and Birch, C.J. 1995. Susceptibility of HIV to inactivation by disinfectants and ultraviolet light. J. Hosp. Inf. 152:400‐403.
   Ericson, J.G., Trevino, A.V., Toedter, G.P., Mathers, L.E., Newbound, G.C., and Lairmore, M.D. 1994. Effects of whole blood lysis and fixation on the infectivity of human T‐lymphotrophic virus type 1 (HTLV‐1). Commun. Clin. Cytometry 18:49‐54.
   Evans, M.R., Henderson, D.K., and Bennett, J.E. 1990. Potential for laboratory exposures to biohazardous agents found in blood. Am. J. Public Health 80:423‐427.
   Ferbas, J., Chadwick, K.R., Logar, A., Patterson, A.E., Gilpin, R.W., and Margolick, J.B. 1995. Assessment of aerosol containment on the ELITE flow cytometer. Cytometry 22:45‐47.
   Ferber, D. 2002. Virology. Monkey virus link to cancer grows stronger. Science 296:1012‐1015.
   Giorgi, J.V. 1994. Cell sorting of biohazardous specimens for assay of immune function. Methods Cell Biol. 42:359‐369.
   Hambleton, P. and Dedonato, G. 1992. Protecting researchers from instrument biohazards. Biotechniques 13:450‐453.
   Harding, L. and Liberman, D.F. 1995. Epidemiology of laboratory‐acquired infections. In Laboratory Safety. Principles and Practices. Second edition. (D.O. Fleming, J.H. Richardson, J.J. Tulis, and D. Vesley, eds.) pp. 7‐15. ASM Press, Washington, D.C.
   Herzenberg, L.A., Parks, D., Sahaf, B., Perez, O., Roederer, M., and Herzenberg, L.A. 2002. The history and future of the fluorescence activated cell sorter and flow cytometry: A view from Stanford. Clin. Chem. 48:1819‐1827.
   Ibrahim, S.F. and van den Engh, G. 2003. High‐speed cell sorting: fundamentals and recent advances. Curr. Opin. Biotechnol. 14:5‐12.
   Ijaz, M.K., Karim, Y.G., Sattar, S.A., and Johnson‐Lussenburg, C.M. 1987. Development of methods to study the survival of airborne viruses. J. Virol. Methods 18:87‐106.
   International Agency for Research on Cancer. 1994. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Hepatitis Viruses. Vol. 59:286. IARC, Lyons, France.
   International Agency for Research on Cancer. 1996. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Human Immunodeficiency Viruses and Human T‐Cell Lymphotropic Viruses. Vol. 67:424. IARC, Lyons, France.
   International Agency for Research on Cancer. 1998. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. Epstein Barr Virus and Kaposi's Sarcoma Herpesvirus/Human Herpesvirus 8. Vol. 70:524. IARC, Lyons, France.
   Jayasinghe, S.M., Wunderlich, J., McKee, A., Newkirk, H., Pope, S., Zhang, J., Staehling‐Hampton, K., Li, L., and Haug, J.S. 2006. Sterile and disposable fluidic subsystem suitable for clinical high speed fluorescence‐activated cell sorting. Cytometry Part B: Clinical Cytometry 70B:344‐354.
   Keane‐Moore, M., Coder, D., and Marti, G. 2002. Public Meeting and Workshop on Safety issues pertaining to the clinical application of flow cytometry to human‐derived cells. Cytotherapy 4:89‐90.
   Leary, J. 2005. Ultra high‐speed sorting. Cytometry Part A 67:76‐85.
   Leemhuis, T. and Adams, D. 2000. Applications of high‐speed sorting for CD34+ hematopoietic stem cells. In Emerging Tools for Single‐Cell Analysis (G. Durack and J.P Robinson, eds.) pp. 73‐93. Wiley‐Liss, New York.
   Lennartz, K., Lu, M., Flasshove, M., Moritz, T., and Kirstein, U. 2005. Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet. Cytometry Part A 66:119‐127.
   Lerche, N.W., Switzer, W.M., Yee, J.L., Shanmugam, V., Rosenthal, A.N., Chapman, L.E., Folks, T.M., and Heneine, W. 2001. Evidence of infection with simian type D retrovirus in persons occupationally exposed to nonhuman primates. J. Virol. 75:1783‐1789.
   Lawrence Livermore National Laboratory. 1995. Biohazardous Operations. Facilities Safety Procedure. 360, Addendum 1. Lawerence Livermore National Laboratory, Livermore, Calif.
   Lopez, P.A. 2002. Basic aspects of high‐speed sorting for clinical applications. Cytotherapy 4:87‐88.
   Martin, L.S., McDougal, J.S., and Loskoski, S.L. 1985. Disinfection and inactivation of the human T lymphotropic virus type III/lymphadenopathy‐associated virus. J. Infect. Dis. 152:400‐403.
   Merrill, J.T. 1981. Evaluation of selected aerosol‐control measures on flow sorters. Cytometry 1:342‐345.
   Mikulich, V.J. and Schriger, D.L. 2002. Abridged version of the updated US Public Health Service guidelines for the management of occupational exposures to hepatitis B virus, hepatitis C virus, and human immunodeficiency virus and recommendations for postexposure prophylaxis. Ann. Emerg. Med. 39:321‐328.
   Musher, D.M. 2003. How contagious are common respiratory tract infections? N. Engl. J. Med. 348:1256‐1266.
   National Institutes of Health. 2002. Notice Pertinent to the April 2002 Revisions of the NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines).
   Oberyszyn, A.S. and Robertson, F.M. 2001. Novel rapid method for visualization of extent and location of aerosol contamination during high‐speed sorting of potentially biohazardous samples. Cytometry 43:217‐222.
   Perfetto, S.P., Ambrozak, D.R., Koup, R.A., and Roederer, M. 2003. Measuring containment of viable infectious cell sorting in high‐velocity cell sorters. Cytometry 52A:122‐130.
   Perfetto, S.P., Ambrozak, D.R., Roederer, M., and Koup, R.A. 2004. Viable infectious cell sorting in a BSL‐3 facility. Methods Mol. Biol. 263:419‐424.
   Rachet, B., Partanen, T., Kauppinen, T., and Sasco, A.J. 2000. Cancer risk in laboratory workers: an emphasis on biological research. Am. J. Ind. Med. 38:651‐665.
   Ruprecht, R., Baba, T.W., Liska, V., Ray, N.B., Martin, L.N., Murphey‐Corb, M., Rizvi, T.A., Bernacky, B.J., Keeling, M.E., McClure, H.M., and Andersen, J., 1999. Oral transmission of primate lentiviruses. J. Infect. Dis. 179:S408‐S412.
   Rutala, W.A. 1996. APIC guidelines for infection control practice. Am. J. Infect. Control 24:313‐342.
   Sattar, S.A. and Ijaz, M.K. 1987. Spread of viral infections by aerosols. Crit. Rev. Environ. Control 89‐131.
   Sattar, S.A. and Springthorpe, V.S. 1991. Survival and disinfectant inactivation of the human immunodeficiency virus: A critical review. Rev. Infect. Dis. 13:430‐447.
   Schmid, I. 2000. Biosafety in the flow cytometry laboratory. In In Living Color, Protocols in Flow Cytometry and Cell Sorting (R.A. Diamond and S. DeMaggio, eds.) pp. 655‐665. Springer Lab Manuals, New York.
   Schmid, I., Nicholson, J.K.A., Giorgi, J.V., Janossy, G., Kunkl, A., Lopez, P.A., Perfetto, S., Seamer, L.C., and Dean, P.N. 1997. Biosafety guidelines for sorting of unfixed cells. Cytometry 28:99‐117.
   Schmid, I., Kunkl, A., and Nicholson, J.K. 1999. Biosafety considerations for flow cytometric analysis of human immunodeficiency virus‐infected samples. Cytometry 38:195‐200.
   Schmid, I., Merlin, S., and Perfetto, S.P. 2003. Biosafety concerns for shared flow cytometry core facilities. Cytometry 56A:113‐119.
   Schmid, I., Roederer, M., Koup, R., Ambrozak, D.R., and Perfetto, S.P. 2004. Biohazard sorting. In Cytometry 4th edition: New Developments. (Z. Darzynkiewicz, M. Roederer, H.J. Tanke, eds.) pp. 221‐240. Elsevier Academic Press, Boston.
   Schoenbaum, M.A., Zimmerman, J.J., Beran, G.W., and Murphy, D.P. 1990. Survival of pseudorabies virus in aerosol. Am .J. Vet. Res. 51:331‐333.
   Schriger, D.L. and Mikulich, V.J. 2002. The management of occupational exposures to blood and body fluids: revised guidelines and new methods of implementation. Ann. Emerg. Med. 39:319‐321.
   Sewell, D.L. 1995. Laboratory‐associated infections and biosafety. Clin. Microbiol. Rev. 8:389‐405.
   Stovel, R.T. 1977. The influence of particles on jet breakoff. J. Histochem. Cytochem. 25:813‐820.
   United States Federal Code Regulation. 1991. Occupational exposure to bloodborne pathogens. CFR PART 1910.1030.
   United States Federal Code Regulation. 2002. Possession, Use, and Transfer of Select Agents and Toxins; Interim Final Rule. 42 CFR Part 1003.
   Van Bueren, J., Simpson, R.A., Salman, H., Farrelly, H.D., and Cookson, B.D. 1995. Inactivation of HIV‐1 by chemical disinfectants: Sodium hypochlorite. Epidemiol. Infect. 115:567‐579.
   Vecchio, D., Sasco, A.J., and Cann, C.I. 2003. Occupational risk in health care and research. Am. J. Ind. Med. 43:369‐397.
   Wang, S.A., Panlilio, A.L., Doi, P.A., White, A.D., Stek, M., Jr., and Saah, A. 2000. Experience of healthcare workers taking postexposure prophylaxis after occupational HIV exposures: findings of the HIV Postexposure Prophylaxis Registry. Infect. Control Hosp. Epidemiol. 21:780‐785.
   Wennborg, H., Yuen, J., Nise, G., Sasco, A.J., Vainio, H., and Gustavsson, P. 2001. Cancer incidence and work place exposure among Swedish biomedical research personnel. Int. Arch. Occup. Environ. Health 74:558‐564.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library