An open-source FACS automation system for high-throughput cell biology
Fig 3
(A) Examples of sorting gates corresponding to the fluorescent tagging of genes with increasing expression level (left to right). As the expression of the fluorescently-tagged gene increases, the gate for the desired engineered population must shift such that only the brightest cells within the sample are collected based on the specific gating algorithm. The same cell samples were sorted both automatically and manually, confirming that the gating algorithm generates results consistent with manual sorting. (B) Automated gating strategy applied in the case study. Contour lines from the fluorescent signal from the cell sample profile (see Fig 1B) are used to define the shape and vertical position of the gate. The gate is translated and rotated such that it includes a user-defined percentage of the fluorescent population (in our case, top 1%). Alternative gating methods can be programmed into the automation control software.