Experiment Overview
| Repository ID: | FR-FCM-ZZYF | Experiment name: | Neutrophil Isolation | MIFlowCyt score: | 51.50% |
| Primary researcher: | Loren Erickson | PI/manager: | Loren Erickson | Uploaded by: | Loren Erickson |
| Experiment dates: | 2011-06-01 - 2012-03-01 | Dataset uploaded: | May 2012 | Last updated: | Jan 2013 |
| Keywords: | [Neutrophil] [autoimmune] [isolation] [spleen] | Manuscripts: | [22760952] [PMC3548322] |
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| Organizations: |
University of Virginia, Charlottesville, VA (USA)
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| Purpose: | The purpose of this experiment was to utilize multiple methods to isolate splenic murine neutrophils and compare and contrast the effectiveness of these protocols based on 3 parameters: purity, viability and activation. We expected that each method would yield a distinct number of viable/pure neutrophils and that each isolation method would result in different degrees of activation of the neutrophils. | ||||
| Conclusion: | In this study, we describe a modified magnetic bead isolation protocol for enriching spleen resident neutrophils by negative selection. EasySep® and Ficoll isolation methods fail to effectively purify neutrophils and particularly, in the case of Ficoll, yield a population of cells with low viability. While sort-purification generates a highly pure and viable population of neutrophils, the process of sorting leads to a substantial activation of the cells. Our results demonstrate that the OAC protocol yields a pure population of neutrophils. Equally important, the OAC protocol does not adversely affect cell viability or activation. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Antibodies were titrated to determine optimal staining conditions. Gates were set based on previously determined FMO. | ||||
