AFCG Biosafety Guidelines for Analysis and Sorting of Unfixed Cells.

ScopeThis document RECOMMENDS procedures to be undertaken during the analysis and sorting of potentially

biohazardous material on

Flow Cytometers.

AFCG website

AFCG Guidelines 2006 - IntroductionPage 2 of guidelines

AFCG website

FOREWORDThese recommendations are presented with a view to being a minimum standard and should not be seen to restrict the ability of any individual. This document will be reviewed regularly to ensure that these recommendations embrace current accepted laboratory practices. These guidelines are written in broad terms as a gesture to indicate and not to dictate.

HOWEVER

EMPLOYER RESPONSIBILITIES

SO FAR AS IS “REASONABLY PRACTICABLE”:

Provide and maintain a safe

o working environment

o plant or systems of work

oworkplace in a condition that is safe and without risk

o system for the use, handling, storage or transport of plant or

substances

WHY ? TO PROTECT THE OPERATOR , THE USERS

AND THE ENVIROMENT.

Biohazardous material

· All unfixed material of human origin.· All unfixed material of primate origin.· All human peripheral blood leukocytes, bone marrow, splenocytes,thymocytes, sperm cells,· Cells from primary and immortalized cultures from humans, primates,and transgenic animals.· All genetically modified organisms (GMO)· Micro-organisms.· Special care to be taken for brain tissue (prions) and lung lavage(tuberculosis).

GENERAL CONSIDERATIONS

1) Policies

GENERAL CONSIDERATIONS

2) BEFORE A NEW USER STARTS:

Flow Cytometry tutorials are mandatory, they cover

theory

practice

and safety.

GENERAL CONSIDERATIONS

3) NOTIFICATION

Implementation

Handling of biohazardous material >> IN TRANSIT AND PREPARATION

STANDARD PC2 page 2 AFCG Guidelines

ImplementationREFERENCES: page 14 of guidelines

· www.ogtr.gov.au/

· Biosafety Guidelines for Sorting of Unfixed Cells, Cytometry, 28:99-117 (1997).

· Cytometry A. 2007 Jun; 71(6):414-37. International Society for Analytical Cytology biosafety standard for sorting of unfixed cells.Schmid I, Lambert C, Ambrozak D, Marti GE, Moss DM, Perfetto SP;International Society of Analytical Cytology.

· Biosafety Considerations for Flow Cytometric Analysis of HIV infected Samples, Cytometry Comm., 38:195-200 (1999).

· Novel Rapid Method for Visualization of Extent and Location of Aerosol Contamination During High-Speed Sorting of Potentially Biohazardous Samples, Cytometry, 42:217-22 (2001).· Measuring Containment of Viable et Infectious Cell Sorting in High-Velocity Cell Sorters. Perfetto et al., Cytometry Part A 52A:122-130(2003)

· Viable Infectious Cell Sorting in a BSL-3 Facility, Flow Cytometry Protocols, vol. 263, Humana Press, 2004

· Biohazard Sorting, Methods in Cell Biology, Cytometry, 4th ed., vol.75,2004Oberyszyn A.S. & Robertson F.M.

.Novel rapid method for visualization of extent and location of aerosol Contamination during high speed sorting of potentially biohazardous samples. Cytometry 2001; 43: 217-222

· Current Protocols in Cytometry Jan 2007: UNIT 3.6 Standard Safety Practices for Sorting of Unfixed Cells. Ingrid Schmid1, Claude Lambert2, David Ambrozak3, and Stephen P. Perfetto3

.Current Protocols in Cytometry Feb 2002 UNIT 3.5 Method for Visualizing Aerosol Contamination in Flow Sorters. Andrew S. Oberyszyn1

.WHO Biosafety Manual.http://www.who.int/csr/resources/publications/biosafety/who

Implementation / Testing

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Implementation / Testing

• “IN HOUSE PROCEDURES”

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SURVEILLANCE

• “IN HOUSE PROCEDURES”> work practices

Ask the question

Passaged cells , how many times

New work , ask what type

Unsure , ask IBC

MANAGEMENT

OHS requirements

IBC Internal Biosafety Committee

AFCG guidelines

Line manager

Decontamination Procedures

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