Experiment Overview
| Repository ID: | FR-FCM-ZZ4Q | Experiment name: | ACCA | MIFlowCyt score: | 63.50% |
| Primary researcher: | Karel Soucek | PI/manager: | Karel Soucek | Uploaded by: | Karel Soucek |
| Experiment dates: | 2010-09-01 - 2012-12-21 | Dataset uploaded: | Dec 2012 | Last updated: | Mar 2013 |
| Keywords: | [clonogenic assay] [clonogenic capacity] [plating efficiency] [cancer stem cells] [extreme limiting dilution analysis] [single cell sorting] | Manuscripts: | [23450810] |
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| Organizations: |
Institute of Biophysics AS CR, v.v.i., Department of Cytokinetics, Brno, (Czech Republic)
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| Purpose: | To establish an automatic cell-cloning assay (ACCA) in a microwell format using an automatic cell deposition unit and a FACS system. | ||||
| Conclusion: | Novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples. | ||||
| Comments: | manuscript submitted to Cytometry part A | ||||
| Funding: | Not disclosed | ||||
| Quality control: | Cell debris, doublets, and aggregates were excluded from analysis based on a dual-parameter dot plot in which the pulse ratio (signal area/signal high; y-axis) versus signal area (x-axis) was displayed. Dead cells were excluded from the analysis. The automatic compensation for multicolor immunofluorescence analysis was performed using BD CompBead beads (BD Biosciences) and particular antibodies. LIVE/DEAD Blue, Yellow, and Far Red dyes (Invitrogen) were compensated using the control sample prepared as a mix of dye-labeled dead cells and negative BD CompBead beads. The sorting rate was no more than 1000 events per second. | ||||
