Table 1 (abstract P-232). Potential sources or error and strategies to control them
Potential Error | Impact | Control strategy |
|---|---|---|
Presence of doublets or clumps of cells | Doublets are unlikely to affect P(monoclonality) as wells with droplets containing these will not be recorded as containing a single cell and will therefore not be progressed. However, presence of a doublet could increase the chance of a cell being sat exactly on top of another and therefore being missed. It will also increase time and resource required to achieve a desired number of wells which both scientists have recorded as containing a single cell. As such, it is good practice to ensure proportion of doublets is low | Ensure cell sample to be cloned contains ≤2% doublets |
Scientists hindered from visualising cells within a droplet | Scientists miss cells and P(monoclonality) is misleadingly high | Procedure in place to exclude wells with droplets harbouring certain characteristics from further analysis and progression |
An error is made in determining the number of cells a droplet contains | P(monoclonality) is misleadingly high or low | (i) Two independent scientists view the droplets (ii) Scientist fatigue is controlled by limiting the amount of time a scientist can spend cloning on any given day (iii) Statistical model takes into account potential errors |