Experiment Overview
| Repository ID: | FR-FCM-ZZYK | Experiment name: | Detection of Cryoglobulins with flowcytometry | MIFlowCyt score: | 86.00% |
| Primary researcher: | Ruediger Mueller | PI/manager: | Ruediger Mueller | Uploaded by: | Ruediger Mueller |
| Experiment dates: | 2007-11-22 - 2012-02-28 | Dataset uploaded: | Jun 2012 | Last updated: | Jan 2013 |
| Keywords: | [flow cytometry] [Cryoglobulins] [detection] | Manuscripts: | |||
| Organizations: |
Kantonsspital St. Gallen, Rheumatology, St. Gallen, St. Gallen (Switzerland)
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| Purpose: | Several patients with cryoglobulin (CG) associated symptoms are seronegative for CG and other potentially causative biomarkers. We analyzed whether it is possible to detect cryoprecipitates by flow cytometry and whether the sensitivity of their demonstration can be increased as compared to visual inspection | ||||
| Conclusion: | Flow cytometry is equally specific but much more sensitive in the detection of CG than visual inspection | ||||
| Comments: | Several patients with cryoglobulin (CG) associated symptoms are seronegative for CG and other potentially causative biomarkers. We analyzed whether it is possible to detect cryoprecipitates by flow cytometry and whether the sensitivity of their demonstration can be increased as compared to visual inspection. Sera from 91 patients with suspected CG associated symptoms and 33 healthy controls were examined for the presence of CG by conventional visual testing and by flow cytometry for small diffracting particles. For calibration purposes we tested lipid micelle dilutions (positive controls) by both methods. The minimum concentrations of lipid micelles to be detected by visual inspection and flow cytometry were 128.5pg/ml and 2.0pg/ml, respectively. Among the 91 patients and 33 controls only one patient serum was positive for CG by conventional testing. This sample was also positive on flow cytometry. In the serum of a patient known to be positive for CG, laser diffracting particles were quantified by flow cytometry after keeping serum at 4°C for 3 days. Of the 91 patients, 14 additional samples displayed cold precipitates which re-dissolved after re-warming during flow cytometry. All fifteen (1 + 14) patients positive for CG on flow cytometry suffered from symptoms usually associated with CG. Some precipitates were labeled with anti IgG and IgM antibodies confirming that the particles detected by flow cytometry contained immunoglobulins. No small diffracting particles were detected in the sera of the 33 healthy controls. Flow cytometry is equally specific but much more sensitive in the detection of CG than visual inspection. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Dataset 2-Flow Cytometer Sensitivity: A Quadratic Model | For calibration purposes we tested lipid micelle dilutions (positive controls) by both methods. The minimum concentrations of lipid micelles to be detected by visual inspection and flow cytometry were 128.5pg/ml and 2.0pg/ml, respectively | |||
Experiment variables
| Conditions | |
|---|---|
| · + cold | CS warm 48h + cold 24h.fcs · EW warm 48h + cold 24h. fcs · HM warm 48h + cold 24h.LMD · NSF warm 48h + cold 24h.LMD · PHB warm 48h + cold 24h.LMD · PIB warm 48h + cold 24h.LMD |
| · + warm | CS cold 48h + warm 24h.fcs · EW cold 48h + warm 24h.fcs · HM cold 48h + warm 24h. fcs · NSF cold 48h + warm 24h.LMD · PHB cold 48h + warm 24h.LMD · PIB cold 48h + warm 24h.LMD |
| · cold 48h | CS cold 48h.fcs · CS warm 48h.fcs · EW cold 48h. fcs · EW warm 48h. fcs · GM cold 48h. fcs · GM warm 48h. fcs · HM cold 48h. fcs · HM warm 48h.LMD · KL cold 48h.LMD · KL warm 48h.LMD · NSF cold 48h.LMD · NSF warm 48h.LMD · PHB cold 48h.LMD · PHB warm 48h.LMD · PIB cold 48h.LMD · PIB warm 48h.LMD |
| · milk | 0 nlMilk-mlPBS. fcs · 1.95 nlMilk-mlPBS. fcs · 1000 nlMilk-mlPBS. fcs · 125 nlMilk-mlPBS. fcs · 15.6 nlMilk-mlPBS.LMD. fcs · 2000 nlMilk-mlPBS. fcs · 250 nlMilk-mlPBS. fcs · 3.9 nlMilk-mlPBS.LMD. fcs · 31.3 nlMilk-mlPBS.LMD. fcs · 4000 nlMilk-mlPBS. fcs · 500 nlMilk-mlPBS. fcs · 5000 nlMilk-mlPBS. fcs · 62.5 nlMilk-mlPBS. fcs · 7.8 nlMilk-mlPBS.LMD. fcs |
| · FCS | 0 nl FCS-mlPBS.fcs · 1000000 nl FCS-mlPBS.fcs · 125000 nl FCS-mlPBS.fcs · 15625 nl FCS-mlPBS. fcs · 250000 nl FCS-mlPBS.fcs · 31250 nl FCS-mlPBS. fcs · 500000 nl FCS-mlPBS.fcs · 62500 nl FCS-mlPBS. fcs · 7813 nl FCS-mlPBS. fcs |
| Individuals | |
|---|---|
| · PHB | PHB cold 48h + warm 24h.LMD · PHB cold 48h.LMD · PHB warm 48h + cold 24h.LMD · PHB warm 48h.LMD |
| · PIB | PIB cold 48h + warm 24h.LMD · PIB cold 48h.LMD · PIB warm 48h + cold 24h.LMD · PIB warm 48h.LMD |
| · NSF | NSF cold 48h + warm 24h.LMD · NSF cold 48h.LMD · NSF warm 48h + cold 24h.LMD · NSF warm 48h.LMD |
| · Milk | 0 nlMilk-mlPBS. fcs · 1.95 nlMilk-mlPBS. fcs · 1000 nlMilk-mlPBS. fcs · 125 nlMilk-mlPBS. fcs · 15.6 nlMilk-mlPBS.LMD. fcs · 2000 nlMilk-mlPBS. fcs · 250 nlMilk-mlPBS. fcs · 3.9 nlMilk-mlPBS.LMD. fcs · 31.3 nlMilk-mlPBS.LMD. fcs · 4000 nlMilk-mlPBS. fcs · 500 nlMilk-mlPBS. fcs · 5000 nlMilk-mlPBS. fcs · 62.5 nlMilk-mlPBS. fcs · 7.8 nlMilk-mlPBS.LMD. fcs |
| · FCS | 0 nl FCS-mlPBS.fcs · 1000000 nl FCS-mlPBS.fcs · 125000 nl FCS-mlPBS.fcs · 15625 nl FCS-mlPBS. fcs · 250000 nl FCS-mlPBS.fcs · 31250 nl FCS-mlPBS. fcs · 500000 nl FCS-mlPBS.fcs · 62500 nl FCS-mlPBS. fcs · 7813 nl FCS-mlPBS. fcs · CS cold 48h + warm 24h.fcs · CS cold 48h.fcs · CS warm 48h + cold 24h.fcs · CS warm 48h.fcs · EW cold 48h + warm 24h.fcs · EW cold 48h. fcs · EW warm 48h + cold 24h. fcs · EW warm 48h. fcs · GM cold 48h. fcs · GM warm 48h. fcs · HM cold 48h + warm 24h. fcs · HM cold 48h. fcs |
| · HM | HM warm 48h + cold 24h.LMD · HM warm 48h.LMD |
| · KL | KL cold 48h.LMD · KL warm 48h.LMD |
| Timepoints | |
|---|---|
| · milk | 0 nlMilk-mlPBS. fcs · 1.95 nlMilk-mlPBS. fcs · 1000 nlMilk-mlPBS. fcs · 125 nlMilk-mlPBS. fcs · 15.6 nlMilk-mlPBS.LMD. fcs · 2000 nlMilk-mlPBS. fcs · 250 nlMilk-mlPBS. fcs · 3.9 nlMilk-mlPBS.LMD. fcs · 31.3 nlMilk-mlPBS.LMD. fcs · 4000 nlMilk-mlPBS. fcs · 500 nlMilk-mlPBS. fcs · 5000 nlMilk-mlPBS. fcs · 62.5 nlMilk-mlPBS. fcs · 7.8 nlMilk-mlPBS.LMD. fcs |
| · nl FCS | 0 nl FCS-mlPBS.fcs · 1000000 nl FCS-mlPBS.fcs · 125000 nl FCS-mlPBS.fcs · 15625 nl FCS-mlPBS. fcs · 250000 nl FCS-mlPBS.fcs · 31250 nl FCS-mlPBS. fcs · 500000 nl FCS-mlPBS.fcs · 62500 nl FCS-mlPBS. fcs · 7813 nl FCS-mlPBS. fcs |
| · + | CS cold 48h + warm 24h.fcs · CS warm 48h + cold 24h.fcs · EW cold 48h + warm 24h.fcs · EW warm 48h + cold 24h. fcs · HM cold 48h + warm 24h. fcs · HM warm 48h + cold 24h.LMD · NSF cold 48h + warm 24h.LMD · NSF warm 48h + cold 24h.LMD · PHB cold 48h + warm 24h.LMD · PHB warm 48h + cold 24h.LMD · PIB cold 48h + warm 24h.LMD · PIB warm 48h + cold 24h.LMD |
| · 48 | CS cold 48h.fcs · CS warm 48h.fcs · EW cold 48h. fcs · EW warm 48h. fcs · GM cold 48h. fcs · GM warm 48h. fcs · HM cold 48h. fcs · HM warm 48h.LMD · KL cold 48h.LMD · KL warm 48h.LMD · NSF cold 48h.LMD · NSF warm 48h.LMD · PHB cold 48h.LMD · PHB warm 48h.LMD · PIB cold 48h.LMD · PIB warm 48h.LMD |
