Literature DB >> 12655656

Measuring containment of viable infectious cell sorting in high-velocity cell sorters.

Stephen P Perfetto1, David R Ambrozak, Richard A Koup, Mario Roederer.   

Abstract

BACKGROUND: With the advent of high-speed sorters, aerosols are a considerable safety concern when sorting viable infectious materials. We describe a four-part safety procedure for validating the containment.
METHODS: This procedure includes aerosol containment, physical barriers, environmental controls, and personal protection. The Aerosol Management System (AMS) produces a negative pressure within the sort chamber, where aerosols are forced through a HEPA filter. Physical barriers include the manufacturer's standard plastic shield and panels. The flow cytometer was contained within a BSL-3 laboratory for maximum environmental control, and the operator was protected by a respiratory system. Containment was measured by using highly fluorescent Glo-Germ particles under the same conditions as the cell sort.
RESULTS: Escaping aerosols were vacuumed for 10 min onto a glass slide and examined. With the AMS active and the cytometer producing the maximum aerosols possible, Glo-Germ particles remained within the sort chamber. Measurements taken directly outside the door averaged fewer than one particle per slide, and those taken at 2 ft away and on top of the sorter were completely negative.
CONCLUSIONS: With this monitoring system in place, aerosols can be efficiently measured, thus reducing the risk to the operator while sorting viable infectious cells. Copyright 2003 Wiley-Liss, Inc.

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Year:  2003        PMID: 12655656     DOI: 10.1002/cyto.a.10033

Source DB:  PubMed          Journal:  Cytometry A        ISSN: 1552-4922            Impact factor:   4.355


  8 in total

1.  Flow cytometry: retrospective, fundamentals and recent instrumentation.

Authors:  Julien Picot; Coralie L Guerin; Caroline Le Van Kim; Chantal M Boulanger
Journal:  Cytotechnology       Date:  2012-01-21       Impact factor: 2.058

Review 2.  Evidence-based biosafety: a review of the principles and effectiveness of microbiological containment measures.

Authors:  Tjeerd G Kimman; Eric Smit; Michèl R Klein
Journal:  Clin Microbiol Rev       Date:  2008-07       Impact factor: 26.132

3.  Novel Impactor and Microsphere-Based Assay Used to Measure Containment of Aerosols Generated in a Flow Cytometer Cell Sorter.

Authors:  Stephen P Perfetto; Phillip J Hogarth; Simon Monard; Ben Fontes; Kristen M Reifel; Brandon K Swan; Jan Baijer; Evan R Jellison; Geoffrey Lyon; Patty Lovelace; Richard Nguyen; David Ambrozak; Kevin L Holmes
Journal:  Cytometry A       Date:  2018-12-18       Impact factor: 4.355

4.  International Society for the Advancement of Cytometry cell sorter biosafety standards.

Authors:  Kevin L Holmes; Benjamin Fontes; Philip Hogarth; Richard Konz; Simon Monard; Charles H Pletcher; Robert B Wadley; Ingrid Schmid; Stephen P Perfetto
Journal:  Cytometry A       Date:  2014-03-13       Impact factor: 4.355

Review 5.  How to develop a standard operating procedure for sorting unfixed cells.

Authors:  Ingrid Schmid
Journal:  Methods       Date:  2012-02-21       Impact factor: 3.608

6.  Characterization of aerosols produced by cell sorters and evaluation of containment.

Authors:  Kevin L Holmes
Journal:  Cytometry A       Date:  2011-11-03       Impact factor: 4.355

7.  Validation of RNA-based molecular clonotype analysis for virus-specific CD8+ T-cells in formaldehyde-fixed specimens isolated from peripheral blood.

Authors:  David van Bockel; David A Price; Tedi E Asher; Vanessa Venturi; Kazuo Suzuki; Kristina Warton; Miles P Davenport; David A Cooper; Daniel C Douek; Anthony D Kelleher
Journal:  J Immunol Methods       Date:  2007-08-09       Impact factor: 2.303

8.  Procedures for Flow Cytometry-Based Sorting of Unfixed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infected Cells and Other Infectious Agents.

Authors:  Kristen M Reifel; Brandon K Swan; Evan R Jellison; David Ambrozak; Jan Baijer; Richard Nguyen; Simon Monard; Geoffrey Lyon; Benjamin Fontes; Stephen P Perfetto
Journal:  Cytometry A       Date:  2020-06-03       Impact factor: 4.355

  8 in total

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