ACCA

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.
Notes:
manuscript submitted to Cytometry part A 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.