Experiment Overview
| Repository ID: | FR-FCM-Z38B | Experiment name: | Sputum_Flow_Analysis | MIFlowCyt score: | 82.46% |
| Primary researcher: | Lydia Bederka | PI/manager: | Vivienne Rebel | Uploaded by: | Lydia Bederka |
| Experiment dates: | 2017-09-01 - 2018-12-31 | Dataset uploaded: | Dec 2020 | Last updated: | Dec 2020 |
| Keywords: | [flow cytometry] [High Throughput] [Leukocytes] [macrophage] [sputum] [alveolar] [interstitial] [diagnostics] | Manuscripts: |
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| Organizations: |
UTHealth San Antonio Flow Cytometry Core Facility, San Antonio , TX (USA)
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| Purpose: | Analysis of the cellular content of sputum via flow cytometry in order to develop a diagnostic platform for lung cancer. | ||||
| Conclusion: | Induced sputum samples were processed to collect single cell suspensions while removing debris through filtration. Sputum samples can be analyzed by flow cytometry while contaminating squamous epithelial cells (SECs) can be excluded from further analysis with the inclusion of a viability dye. Two macrophage populations were identified, and morphologies confirmed by sorting experiments, that can serve as verification that the sample was obtained from the lung as opposed to being saliva. We determined at least 0.75 million cells are needed for reliable analysis. The use of flow cytometry to analyze sputum-derived cells serves as a reliable, non-invasive means to monitor the health status of the lung. | ||||
| Comments: | Three-day sputum collection from persons aged 55-74 with at least a 30 pack-year smoking history at high risk of developing lung cancer. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | LSR II cytometer was checked prior to each analysis session with BD FACSDiva™ CS&T Research beads. The LSR II PMTs are adjusted and checked prior to each use using SPHERO™ Rainbow Calibration Particles. | ||||
Experiment variables
| Conditions | |
|---|---|
| · Compensation | A549 FVS510.fcs · A549 TCPP.fcs · A549 unstained.fcs · FITC Comp Beads.fcs · PE Comp Beads.fcs · PE TexRed Comp Beads.fcs · PE COMP Beads.fcs · PE-CF594 COMP Beads.fcs · PE Comp Beads.fcs-(1) · PE-CF594 Comp Beads.fcs · A549 + 5X FVS-510.fcs · A549 + TCPP.fcs · A549 unstained.fcs-(1) · Specimen_001_A549 FVS510_007.fcs · Specimen_001_A549 TCPP_008.fcs · Specimen_001_A549 unstained_006.fcs · Specimen_001_FITC comp beads_004.fcs · Specimen_001_PE comp beads_003.fcs |
| · Sputum-Unstained | BL-18-167_unstained.fcs · BL-18-179 Unstained.fcs · BL-18-188 unstained.fcs · BL-18-084 Unstained.fcs · Specimen_001_BL-18-146 unstained_011.fcs |
| · Sputum-Isotype | BL-18-167_isotype.fcs · BL-18-179 Isotype.fcs · BL-18-188 isotype.fcs · BL-18-084 - Isotypes.fcs · Specimen_001_BL-18-146 CD45 + isotype_014.fcs |
| · Sputum-Blood | BL-18-167_Blood.fcs · BL-18-179 Blood.fcs · BL-18-188 Blood.fcs · BL-18-084 - Blood.fcs · Specimen_001_BL-18-146 Blood_017.fcs |
