Experiment Overview
| Repository ID: | FR-FCM-ZYXU | Experiment name: | Phagocytosis of A.fumigatus conidia | MIFlowCyt score: | 81.31% |
| Primary researcher: | Susann Hartung | PI/manager: | Susann Hartung | Uploaded by: | Susann Hartung |
| Experiment dates: | 2015-09-04 - 2015-09-04 | Dataset uploaded: | Aug 2018 | Last updated: | Mar 2019 |
| Keywords: | [phagocytosis] [Aspergillus fumigatus] [anti-FITC] | Manuscripts: | [30450827] |
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| Organizations: |
University Hospital Jena, Hematology and Oncology, Jena, (Germany)
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| Purpose: | We were investigating phagocytosis of Aspergillus fumigatus conidia by primary human neutrophils and monocytes using a novel technique. FITC-labelled conidia were allowed to be phagocytosed and then stained with an APC-labelled anti-FITC antibody to allow for discrimination of cells with internalised conidia (FITC+ APC-) or adherent conidia (FITC+ APC+). This assay was done for different time points using either resting / dormant or pre-swollen conidia. | ||||
| Conclusion: | Our proposed method to analyze phagocytosis of FITC-labeled conidia by human primary phagocytes and to distinguish internalization and adherence by counterstaining with an anti-FITC antibody is an easy, reliable and flexible way to quantify host-pathogen interaction. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Samples with conidia were controlled by samples without conidia (unstimulated). FITC labeled conidia are controlled by unlabeled conidia. CST beads were used to check cytometer performance regularly. | ||||
Experiment variables
| Timepoints | |
|---|---|
| · 0.5h | IV_Inkubation 0_5h_2_5h geschwollen FITC_007.fcs · IV_Inkubation 0_5h_4+1h geschwollen FITC_008.fcs · IV_Inkubation 0_5h_RPMI_005.fcs · IV_Inkubation 0_5h_resting FITC_006.fcs · ungefaerbt_0_5h_2_5h geschwollen FITC_003.fcs · ungefaerbt_0_5h_4+1h geschwollen FITC_004.fcs · ungefaerbt_0_5h_RPMI_001.fcs · ungefaerbt_0_5h_resting FITC_002.fcs |
| · 2h | IV_Inkubation 2h_2_5h geschwollen FITC_011.fcs · IV_Inkubation 2h_4+1h geschwollen FITC_012.fcs · IV_Inkubation 2h_RPMI_009.fcs · IV_Inkubation 2h_resting FITC_010.fcs · ungefaerbt_2h_2_5h geschwollen FITC_020.fcs · ungefaerbt_2h_4+1h geschwollen FITC_021.fcs · ungefaerbt_2h_RPMI_018.fcs · ungefaerbt_2h_resting FITC_019.fcs |
| · 4h | IV_Inkubation 4h_2_5h geschwollen FITC_015.fcs · IV_Inkubation 4h_4+1h geschwollen FITC_017.fcs · IV_Inkubation 4h_4+1h geschwollen ohne_016.fcs · IV_Inkubation 4h_RPMI_013.fcs · IV_Inkubation 4h_resting FITC_014.fcs · komplett ungefaerbt 4h_Konidien FITC 4+1h_029.fcs · komplett ungefaerbt 4h_Konidien unmarkiert 4+1h_028.fcs · komplett ungefaerbt 4h_RPMI_027.fcs · ungefaerbt_4h_2_5h geschwollen FITC_024.fcs · ungefaerbt_4h_4+1h geschwollen FITC_026.fcs · ungefaerbt_4h_4+1h geschwollen ohne_025.fcs · ungefaerbt_4h_RPMI_022.fcs · ungefaerbt_4h_resting FITC_023.fcs |
| Conditions | |
|---|---|
| · resting | IV_Inkubation 0_5h_resting FITC_006.fcs · IV_Inkubation 2h_resting FITC_010.fcs · IV_Inkubation 4h_resting FITC_014.fcs · ungefaerbt_0_5h_resting FITC_002.fcs · ungefaerbt_2h_resting FITC_019.fcs · ungefaerbt_4h_resting FITC_023.fcs |
| · 2_5h swollen | IV_Inkubation 0_5h_2_5h geschwollen FITC_007.fcs · IV_Inkubation 2h_2_5h geschwollen FITC_011.fcs · IV_Inkubation 4h_2_5h geschwollen FITC_015.fcs · ungefaerbt_0_5h_2_5h geschwollen FITC_003.fcs · ungefaerbt_2h_2_5h geschwollen FITC_020.fcs · ungefaerbt_4h_2_5h geschwollen FITC_024.fcs |
| · 5h swollen | IV_Inkubation 0_5h_4+1h geschwollen FITC_008.fcs · IV_Inkubation 2h_4+1h geschwollen FITC_012.fcs · IV_Inkubation 4h_4+1h geschwollen FITC_017.fcs · ungefaerbt_0_5h_4+1h geschwollen FITC_004.fcs · ungefaerbt_2h_4+1h geschwollen FITC_021.fcs · ungefaerbt_4h_4+1h geschwollen FITC_026.fcs |
| · unstimulated | IV_Inkubation 0_5h_RPMI_005.fcs · IV_Inkubation 2h_RPMI_009.fcs · IV_Inkubation 4h_RPMI_013.fcs · ungefaerbt_0_5h_RPMI_001.fcs · ungefaerbt_2h_RPMI_018.fcs · ungefaerbt_4h_RPMI_022.fcs |
| · no FITC 5h swollen | IV_Inkubation 4h_4+1h geschwollen ohne_016.fcs · ungefaerbt_4h_4+1h geschwollen ohne_025.fcs |
| · no staining | komplett ungefaerbt 4h_Konidien FITC 4+1h_029.fcs · komplett ungefaerbt 4h_Konidien unmarkiert 4+1h_028.fcs · komplett ungefaerbt 4h_RPMI_027.fcs |
