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Figure 2 | BMC Research Notes

Figure 2

From: Optimizing cell arrays for accurate functional genomics

Figure 2

Quantitative analysis of array accuracy. (a) Schematic representation of automated image analysis. Schematic of the module built pipeline created with Cell Profiler software to calculate the numbers of mCitrine and mCherry expressing cells in mCitrine spots. Thresholds for mCitrine and mCherry channels were applied as described in methods. The total number of cells was calculated by identifying primary objects by using a nuclei staining. The threshold images of mCitrine or mCherry were used with the primary objects to identify mCitrine and mCherry expressing cells (secondary objects). The white arrow shows one of the interspot contaminating cells. (b) and (c) The percentage of mCitrine expressing cells was calculated for each mCitrine cDNA-containing spot (transfection efficiency, back bars) or for mCherry cDNA-containing spots (interspot contamination, light-gray bars). Dark-grey bars represent the % of mCitrine-expressing cells in empty areas. Results obtained with the protocol that includes the washing step are shown. An example of cropped areas used for 20x objective simulation is shown (left panels). The number of mCitrine cells and the standard deviation are indicated.

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