Experiment Overview
| Repository ID: | FR-FCM-Z5L8 | Experiment name: | Doublets | MIFlowCyt score: | 84.00% |
| Primary researcher: | Jordi Petriz | PI/manager: | Jordi Petriz | Uploaded by: | Jordi Petriz |
| Experiment dates: | 2017-04-24 - 2017-05-25 | Dataset uploaded: | Jul 2022 | Last updated: | Jul 2022 |
| Keywords: | [flow cytometry] [light scatter] [doublet discrimination] [viable DNA dyes] | Manuscripts: | |||
| Organizations: |
Germans Trias i Pujol Research Institute, Functional Cytomics, Badalona, Barcelona (Spain)
|
||||
| Purpose: | To compare the use of light scattering with fluorescence measurement techniques for successful doublet discrimination for single cells by using peripheral blood samples were prepared with and without fixative and non-fixative erythrocyte lysing solutions. | ||||
| Conclusion: | - The obtention of single cell preparations is crucial to characterize cell suspensions, even obtained after tissue and tumor disaggregation for flow cytometric analysis, especially when is extremely needed to detect rare events, by maximizing the signal to noise ratio and by increasing the number of acquired events. - Overall, fluorescent methods for thresholding and doublet discrimination can be very helpful to identify coincident events and to increase the resolution of singlets. - Doublet discrimination by fluorescence takes advantage not only for NLNW methods, because also simplifies the identification of aggregates, by avoiding artifacts during sample manipulation, as well as the use of inaccurate light scatter triggering for small pathological cells or early apoptotic events as a consequence of cell shrinkage. | ||||
| Comments: | None | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Performance Tracking Beads (daily) | ||||
Experiment variables
| Conditions | |
|---|---|
| · No Lysis | 2017_172 BLOOD16 NoLyse CE.fcs · 2017_172 BLOOD16 NoLyse NCE.fcs |
| · Ammonium Chloride | BLOOD16 AC CE(1).fcs · BLOOD16 AC NCE.fcs |
| · CyLyse | BLOOD16 CyLyse CE.fcs · BLOOD16 CyLyse NCE.fcs |
| · CD34 staining | APH CD34 CD45 rep11.fcs |
| Individuals | |
|---|---|
| · healthy donnors | 2017_172 BLOOD16 NoLyse CE.fcs · 2017_172 BLOOD16 NoLyse NCE.fcs · APH CD34 CD45 rep11.fcs · BLOOD16 AC CE(1).fcs · BLOOD16 AC NCE.fcs · BLOOD16 CyLyse CE.fcs · BLOOD16 CyLyse NCE.fcs |
| Sample Type | |
|---|---|
| · peripheral blood | 2017_172 BLOOD16 NoLyse CE.fcs · 2017_172 BLOOD16 NoLyse NCE.fcs · BLOOD16 AC CE(1).fcs · BLOOD16 AC NCE.fcs · BLOOD16 CyLyse CE.fcs · BLOOD16 CyLyse NCE.fcs |
| · apheresis product | APH CD34 CD45 rep11.fcs |
