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. 2013 May 14;8(5):e63286. doi: 10.1371/journal.pone.0063286

Figure 3. Improved well-to-well reproducibility in cell growth measurement enabled by the FPNLSFP live cell nuclear marker.

Figure 3

Variations in cell numbers plated in each well (Day 0) obscured the ability to reliably detect an increase in cell number after four days of slow growth by LNCaP-C4-2 prostate cancer cells treated with vehicle or 0.2 nM DHT. Each symbol represents the numbers of A, Hoechst 33342-stained nuclei or B, FPNLSFP-marked nuclei segmented in each well. C, Dividing the number of FPNLSFP-marked cells on Day 4 by the baseline (Day 0) number of FPNLSFP-marked cells in the same well improved the reproducibility of growth measurement. Dotted lines, three standard deviations (3sd) above the mean Day 0 (black dotted line) or Day 4 vehicle-treated (gray dotted line) measurements are shown. The 3sd cut-offs were used to determine the number of, respectively, vehicle-treated and DHT-treated wells that were scored falsely in the Day 0 and vehicle-treated wells.