The Focinator v2-0 - Graphical Interface, Four Channels, Colocalization Analysis and Cell Phase Identification.

Area of Science:

• Cell Biology
• Radiation Biology
• Experimental Oncology

Background:

• Quantitative analysis of DNA damage foci is crucial for understanding radiation biology and oncology.
• Key proteins like γ-H2A.X, 53BP1, and RAD51 form repair foci at DNA damage sites.
• Automated tools are needed to efficiently analyze these foci.

Purpose of the Study:

• To refine and enhance "The Focinator" software for automated analysis of nuclei and DNA damage foci.
• To improve user-friendliness and introduce advanced features for quantitative and qualitative analysis.

Main Methods:

• Development of a refined software tool, "The Focinator v2.0", with a graphical interface.
• Implementation of an R-script-based mode for automated image processing, file management, and error reporting.
• Inclusion of multi-channel analysis (up to four channels) and protein-protein colocalization assessment (up to three foci channels).

Main Results:

• The Focinator v2.0 provides a more user-friendly experience with automated operations, reducing the need for constant operator supervision.
• The software enables advanced multi-channel analysis and colocalization studies.
• Automated image opening, file naming, progress monitoring, and error reporting are now integrated.

Conclusions:

• The Focinator v2.0 significantly improves the efficiency and scope of DNA damage foci analysis in cell biology research.
• The enhanced software facilitates complex analyses, such as quantifying foci in specific cell cycle phases.
• This tool supports researchers in radiation biology and experimental oncology by providing reliable and fast automated analysis.