Experiment Overview
| Repository ID: | FR-FCM-Z2ZF | Experiment name: | Antibody Cocktail Freezing - Full immune phenotyping cocktail | MIFlowCyt score: | 79.75% |
| Primary researcher: | Axel Schulz | PI/manager: | Axel Schulz | Uploaded by: | Axel Schulz |
| Experiment dates: | 2016-10-19 - 2017-02-28 | Dataset uploaded: | Mar 2019 | Last updated: | Mar 2019 |
| Keywords: | [mass cytometry] [CyTOF] [Standardization] [antibody cocktail preservation] | Manuscripts: | [31058420] |
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| Organizations: |
Deutsches Rheuma-Forschungszentrum Berlin, Flow Cytometry Core Facility (FCCF), Berlin, (Germany)
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| Purpose: | Test whether cocktail cryopreservation for mass cytometry works also on a complex immune phenotyping panel at a storage temperature of -80°C. Test compatibilty with commercial and in-house conjugates that contain PBS-based antibody stabilizing solution (Candor) | ||||
| Conclusion: | Cryopreserved cocktails stored at -80°C for 6 or 19 weeks created similar staining patterns and signal intensities when compared to the same cocktail used before freezing to stain the same PBMC, and highly reproducible composition and clustering of immune cell populations in t-SNE maps. Minor variability in staining intensities across the different storage times remained, but again did not correlate with the length of cocktail storage. For example, CD14-160Gd showed highest specific signal intensity after 19 weeks of storage without increased background. For CD28-161Dy, the highest signal was observed for the untreated cocktail and the lowest after 6 weeks of storage. CD45 labeled with 198Pt did not show notable differences across the test conditions, while CD45 labeled with 110Pd showed a slightly increased signal after storage for 19 weeks compared to fresh cocktail and 6 weeks of storage | ||||
| Comments: | Refers to Fig 2 and Fig. S2 | ||||
| Funding: | DFG Me-3644/5-1, SCHU-3263/1-1, TRR130 TP24 RTCure, #777357, DRFZ, Else-Kröner-Fresenius-Stiftung. | ||||
| Quality control: | 4 elements EQ beads Data is already normalized (Fluidigm algorithm) | ||||
Experiment variables
| Conditions | |
|---|---|
| · fresh | export_AxS_20161020_TSORATest_ND031_1_time_Ungated_viSNE.fcs |
| · -80°C_6wk | export_AxS_20161129_TSORA_frozenMM_0_time_Ungated_viSNE.fcs |
| · -80°C_19wk | export_AxS_20170228_TSORA_-80degreeMM_0_time_Ungated_viSNE.fcs |
