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Table 1 (abstract P-232). Potential sources or error and strategies to control them

From: Abstracts from the 25th European Society for Animal Cell Technology Meeting: Cell Technologies for Innovative Therapies

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