Fluorescent-activated cell sorting (FACS) is often the most appropriate technique to obtain pure populations of a cell type of interest for downstream analysis. However, aerosol droplets can be generated during the sort, which poses a biosafety risk when working with samples containing risk group 3 pathogens such as , , , and severe acute respiratory syndrome coronavirus 2. For many researchers, placing the equipment required for FACS at biosafety level 3 (BSL-3) is often not possible due to expense, space, or expertise available. We performed aerosol testing as part of the biosafety evaluation of the MACSQuant Tyto, a completely closed, cartridge-based cell sorter. We also established quality control procedures to routinely evaluate instrument performance. The MACSQuant Tyto does not produce aerosols as part of the sort procedure. These data serve as guidance for other facilities with containment laboratories wishing to use the MACSQuant Tyto for cell sorting. Potential users should consult with their Institutional Biosafety Committees to perform in-house risk assessments of this equipment. The MACSQuant Tyto can safely be used on the benchtop to sort samples at BSL-3.
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http://dx.doi.org/10.1089/apb.20.0065 | DOI Listing |
STAR Protoc
December 2024
Division of Experimental Allergy and Immunodermatology, School of Medicine and Health Sciences, Carl von Ossietzky University Oldenburg, Oldenburg, Germany; Research Center for Neurosensory Science, Carl von Ossietzky University Oldenburg, Oldenburg, Germany; University Clinic of Dermatology and Allergy, University of Oldenburg, Oldenburg, Germany. Electronic address:
Most common techniques for isolating eosinophils utilize CD16-negative selection, neglecting the CD16-positive fraction of eosinophils. Here, we present a protocol for isolating human CD16+ and CD16- eosinophils based on their autofluorescence using the MACSQuant Tyto cell sorter. We describe steps for purifying eosinophils and assessing purity, viability, and functional activity.
View Article and Find Full Text PDFJ Transl Med
November 2024
Department of Myxoma, Aerospace Center Hospital, Beijing, 100049, China.
Background: Pseudomyxoma peritonei (PMP) is a rare malignant peritoneal tumor that readily recurs and metastasizes. Studies have shown that cancer stem cells (CSCs) play an important role in tumor recurrence, metastasis, and prognosis.
Objective: In this study, our aim was to isolate CSCs from various tissues of PMP patients and compare their proliferation, migration, and anti-inflammatory abilities.
Appl Biosaf
December 2021
Immunity to Pulmonary Pathogens Section, Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA.
Fluorescent-activated cell sorting (FACS) is often the most appropriate technique to obtain pure populations of a cell type of interest for downstream analysis. However, aerosol droplets can be generated during the sort, which poses a biosafety risk when working with samples containing risk group 3 pathogens such as , , , and severe acute respiratory syndrome coronavirus 2. For many researchers, placing the equipment required for FACS at biosafety level 3 (BSL-3) is often not possible due to expense, space, or expertise available.
View Article and Find Full Text PDFCytometry A
April 2022
Department of Biomedicine, University of Aarhus, Aarhus C, Denmark.
Isolation of multiple cell populations from limited starting material and with minimal influence on cell homeostasis and viability are common requirements in both basic and clinical research. Fluorescence-activated cell sorting (FACS) is the most commonly applied sorting methodology with the majority of instruments being based on high pressure and electrostatic deflection. A more recent technology is based on a mechanical valve, operating at low pressure.
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