Experiment Overview
| Repository ID: | FR-FCM-ZZP8 | Experiment name: | Assessment of stability of the flow cytometric microbiota profile prior to formaldehyde fixation | MIFlowCyt score: | 88.51% |
| Primary researcher: | Jakob Zimmermann | PI/manager: | Hyun-Dong Chang | Uploaded by: | Jakob Zimmermann |
| Experiment dates: | 2014-08-19 - 2014-08-19 | Dataset uploaded: | Feb 2016 | Last updated: | Feb 2016 |
| Keywords: | [flow cytometry] [microbial communities] [Microbiota] | Manuscripts: | [26909672] |
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| Organizations: |
Helmholtz-Centre for Environmental Research - UFZ, Env. microbiology, Leipzig, (Germany)
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| Purpose: | If multiple samples for microbiota analysis are collected, storage on ice prior to formaldehyde fixation is inevitable. It has been shown that the flow-cytometric FSC/DAPI profile of bacterial communities remains stable long-term after formaldehyde fixation and storage in 70% EtOH at -20°C (REF). In this experiment, it was addressed how storage time prior to formaldehyde fixation affects the FSC/DAPI profile of fecal microbiota samples. | ||||
| Conclusion: | The FSC/DAPI profile of samples remains stable for at least 3 hours prior to formaldehyde fixation. Thus, multiple samples of fecal microbiota can be collected at once and be stored on ice for at least 3 hours without affecting their FSC/DAPI profile. | ||||
| Comments: | None | ||||
| Funding: | This work was supported by the German Research Council (DFG SFB 633, SFB 650, SPP 1656, and GRK1121) and the European Research Council advanced grant ERC-2010-AdG_20100317 Grant 268987. The DRFZ is a Leibniz institute. | ||||
| Quality control: | Fluorescent beads FluoSpheres 2 µm (505/515, Lot-Nr. 24498W, Molecular Probes Eugene, Oregon, USA), FluoSpheres 1 µm (350/440, Lot-Nr. 69A1-1, Molecular Probes Eugene, Oregon, USA), and Fluoresbrite BB Carboxylate microspheres 0.5 µm (360/407, Lot-Nr. 552744, Polyscience, USA) were used to align the instrumental settings. | ||||
Experiment variables
| Timepoints | |
|---|---|
| · 15 | PBS1_15.fcs · PBS2_15.fcs |
| · 30 | PBS1_30.fcs · PBS2_30.fcs |
| · 45 | PBS1_45.fcs · PBS2_45.fcs |
| · 60 | PBS1_60.fcs · PBS2_60.fcs |
| · 120 | PBS1_120.fcs · PBS2_120.fcs |
| · 180 | PBS1_180.fcs · PBS2_180.fcs |
| · 0 | PBS1_0.fcs · PBS2_0.fcs |
| Individuals | |
|---|---|
| · 1 | PBS1_0.fcs · PBS1_120.fcs · PBS1_15.fcs · PBS1_180.fcs · PBS1_30.fcs · PBS1_45.fcs · PBS1_60.fcs |
| · 2 | PBS2_0.fcs · PBS2_120.fcs · PBS2_15.fcs · PBS2_180.fcs · PBS2_30.fcs · PBS2_45.fcs · PBS2_60.fcs |
