Fig. 4
From: The Focinator - a new open-source tool for high-throughput foci evaluation of DNA damage
Use of the Focinator macro reduces counting times compared to ImageJ-based counting and manual evaluation. TRAMP-C1 cells were irradiated with 3 Gy. The cells were fixed and permeabilized for 15 min with 3 % PFA and 0.2 % Triton X-100 at different time points after irradiation. The nuclei were stained with Hoechst 33342. DSB foci were labeled with Alexa Fluor 647-linked anti- γ-H2.AX antibodies. The evaluation time for the same 35 multi-channel images containing 439 nuclei was compared between the analysis with the Focinator, ImageJ-based counting via manual ROI marking and “Find Maxima…” function or manual counting. a Evaluation times using the different counting methods. b Comparison of detected nuclei numbers by ImageJ-based analysis, Focinator batch mode and manual counting shown as overall ROI count