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Lab Safety Manual

Environmental Health and Safety
108 Edwards Hall
1810 Kerr Dr.
Kansas State University
Manhattan, KS 66506

785-532-5856
785-532-1981 fax
safety@k-state.edu

Cell Sorters and Cytometers

Flow cytometers are automated instruments that provide quantitative measures of the properties of single cells using laser based techniques. Operators of these instruments are therefore at risk for biohazard and laser exposure.

Sample identification and handling

Hazard identification is an important step in risk analysis and informs the selection of safety protocol. For this reason it is important for the operator to have specific knowledge of the nature of the samples being handled.

  1. Procedures should be developed to identify the potential biological hazards of a specimen and for containment of aerosols, waste management, and equipment maintenance.
  2. Biosafety information about a sample should be collected prior to the analysis of a given sample.
  3. This information should include: origin of sample (human, cultured cell lines, rodent, etc.), pathological screening information, use of viral vectors, have the infectious agents been inactivated, presence of oncogenes, presence of genetic modification, etc.
  4. An appropriate BSL and associated procedure can then be implemented.
Aerosol containment

Risk of exposure to biological material primarily arises from sample handling and the potential generation of aerosols during analysis or during sample preparation. Biological specimens can contain known or unknown pathogens and therefore should be handled with care. Aerosol containment measures should be taken if a specimen is identified as infectious or potentially infectious.

  1. Cytometers analyzing or sorting infectious materials or unfixed biological specimens must be placed within a BSC or other suitable containment device.
  2. Only fixed materials may be sorted or analyzed in a cytometer outside of a BSC.
  3. If the sample contains any genetically engineered amphotropic virus, it must be treated as infectious and BSL-2 practices must be implemented.
  4. Vortexing biological specimens in open tubes is prohibited.
  5. Vortexing closed tubes should take place in a BSC.
  6. Samples modified using viral vectors to introduce hazardous elements into the cells may not be sorted or analyzed outside of a BSC.
  7. All handling of unfixed human tissue must be done in accordance with the directions under the Bloodborne Pathogens section of the Laboratory Safety Manual.
Laser safety

Lasers are a common component in cytometry instruments. These components are generally protected by safety devices. A Cytometer only poses a laser hazard when these safety have been disengaged or otherwise defeated. For additional information, see Laser Safety.

Maintenance

Proper maintenance and record keeping is vital to insuring instruments are working properly. Engineering controls offer a high level of protection, but only when working properly. Filters require routine change outs, flow rates can change based on laboratory conditions and may need to be adjusted.

  1. Efficiency of aerosol containment should be tested periodically. Records of these tests should be kept.
  2. Operating manuals should be used to guide instrument maintenance and scheduling.
  3. Waste effluent shall be captured in a container with concentrated bleach such that the final bleach concentration is 10%
  4. Fluid lines shall be routinely decontaminated with a <10% bleach solution
  5. Dispose of waste according to hazardous waste disposal guidelines.