Fig. 5
From: The Focinator - a new open-source tool for high-throughput foci evaluation of DNA damage
The Focinator’s accuracy is comparable to manual counting and evaluation only with ImageJ. ImageJ-based, manual counting and the usage of the Focinator macro were compared. To evaluate the repair time-dependent decrease of γ-H2.AX foci after irradiation TRAMP-C1 cells were irradiated with 3 Gy, incubated at 37 °C and fixed 0.5, 1, 2, 4, 6 and 24 h after irradiation. The cells were permeabilized and stained with an Alexa 647-linked anti- γ-H2.AX antibody. A total number of approximately 40 nuclei per time point was evaluated. a Development of the mean foci count per nucleus form three independent experiments at stated time points after irradiation. b A dose response curve depicts foci count after different doses (0.5, 1.5 and 3 Gy) 30 min after irradiation. A direct correlation between the different scoring methods with respective correlations value (R2) at the time points 0.5, 1, 2, 4, 6 and 24 h after irradiation is shown for Focinator-based evaluation in comparison to using ImageJ alone in (c) and compared to manual counting in (d)