*4.10. Images Quantification and Registration*

Cells were seeded into 6-well plates and allowed to grow for 48 h before treatment. The cells were fixed, permeabilized and stained for γ-H2AX or Rad51 by using specific antibodies. Plates were imaged with a × 10 objective using a Cytell Cell Imaging System (GE Healthcare, Chicago, Illinois, USA). MyBioApp Protocol with specified parameters was created to acquire the data and to quantify the signal intensity in nuclei, cytoplasm, and in whole cells. DAPI (blue channel) was used for the nuclear masks, whereas Alexa Fluor 647 Mouse (red channel) was used for γ-H2AX-staining and Alexa Fluor 647 Rabbit (red channel) for Rad51-staining. Graphics illustrating the average intensity of the nuclear γ-H2AX-staining and Rad51-staining were automatically generated by using MyBioApp Protocol and then passed to MS Excel for further processing and analysis.
