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Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet.
Cytometry A. 2005 Aug; 66(2):119-27.C

Abstract

BACKGROUND

The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2.

METHODS

After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions.

RESULTS

The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabinet, does not influence the conditions for side stream stability or the analytical precision of the FACS Vantage cell sorting system. In addition, tests performed to assess aerosol containment during operation of the special biosafety cabinet demonstrated that the cabinet-integrated FACS Vantage unit (CIF) satisfies the conditions for class II cabinets. In the context of gene transfer experiments, the CIF facility was used to sort hematopoietic progenitor cells under biosafety level 2 conditions.

CONCLUSIONS

The newly designed biosafety cabinet offers a practical modality for improving biosafety for operators and samples during cell sorting procedures. It can thus also be used for sorting experiments with genetically modified organisms in conformance with current biosafety regulations and guidelines.

Authors+Show Affiliations

Lennartz K
Institute of Cell Biology (Cancer Research), West German Cancer Center Essen, University of Duisburg-Essen, Medical School, Essen, Germany.
Lu M
No affiliation info available
Flasshove M
No affiliation info available
Moritz T
No affiliation info available
Kirstein U
No affiliation info available

MeSH

AerosolsAir MovementsAntigens, CD34Cell SeparationContainment of BiohazardsCytidine DeaminaseEnvironment, ControlledEquipment ContaminationEquipment DesignFlow CytometryGreen Fluorescent ProteinsGuidelines as TopicHazardous SubstancesHematopoietic Stem CellsHumansOccupational HealthOrganisms, Genetically ModifiedRetroviridaeSafety ManagementTransduction, Genetic

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15973698

Citation

Lennartz, Klaus, et al. "Improving the Biosafety of Cell Sorting By Adaptation of a Cell Sorting System to a Biosafety Cabinet." Cytometry. Part a : the Journal of the International Society for Analytical Cytology, vol. 66, no. 2, 2005, pp. 119-27.
Lennartz K, Lu M, Flasshove M, et al. Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet. Cytometry A. 2005;66(2):119-27.
Lennartz, K., Lu, M., Flasshove, M., Moritz, T., & Kirstein, U. (2005). Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet. Cytometry. Part a : the Journal of the International Society for Analytical Cytology, 66(2), 119-27.
Lennartz K, et al. Improving the Biosafety of Cell Sorting By Adaptation of a Cell Sorting System to a Biosafety Cabinet. Cytometry A. 2005;66(2):119-27. PubMed PMID: 15973698.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet. AU - Lennartz,Klaus, AU - Lu,Mengji, AU - Flasshove,Michael, AU - Moritz,Thomas, AU - Kirstein,Uwe, PY - 2005/6/24/pubmed PY - 2005/10/21/medline PY - 2005/6/24/entrez SP - 119 EP - 27 JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology JO - Cytometry A VL - 66 IS - 2 N2 - BACKGROUND: The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2. METHODS: After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions. RESULTS: The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabinet, does not influence the conditions for side stream stability or the analytical precision of the FACS Vantage cell sorting system. In addition, tests performed to assess aerosol containment during operation of the special biosafety cabinet demonstrated that the cabinet-integrated FACS Vantage unit (CIF) satisfies the conditions for class II cabinets. In the context of gene transfer experiments, the CIF facility was used to sort hematopoietic progenitor cells under biosafety level 2 conditions. CONCLUSIONS: The newly designed biosafety cabinet offers a practical modality for improving biosafety for operators and samples during cell sorting procedures. It can thus also be used for sorting experiments with genetically modified organisms in conformance with current biosafety regulations and guidelines. SN - 1552-4922 UR - https://www.unboundmedicine.com/medline/citation/15973698/Improving_the_biosafety_of_cell_sorting_by_adaptation_of_a_cell_sorting_system_to_a_biosafety_cabinet_ DB - PRIME DP - Unbound Medicine ER -