Automated cell segmentation in FIJI® using the DRAQ5 nuclear dye

Area of Science:

• Cellular analysis
• Quantitative microscopy
• Biotechnology applications

Background:

• Image segmentation and quantification are crucial for cellular analysis.
• Existing methods can be complex for non-specialists.
• There is a need for accessible, automated tools.

Purpose of the Study:

• To develop a fast, customizable, and unsupervised cell segmentation method.
• To utilize Fiji software and DRAQ5 fluorescence for automated analysis.
• To enable quantitative cellular analysis for non-image processing specialists.

Main Methods:

• Developed an unsupervised cell segmentation method using Fiji (ImageJ).
• Employed "leaky" fluorescence from the DNA stain DRAQ5 for nucleus detection.
• Applied the method for 2D cell segmentation and quantitative analysis.

Main Results:

• Achieved >92% accuracy and 94% sensitivity in HeLa cell segmentation.
• Reported 86% correct segmentation and IoU score >0.83.
• Quantified cell area changes in differentiating THP-1 cells and temperature-treated CHO cells.

Conclusions:

• The developed method is straightforward and easily applicable.
• The staining protocol is user-friendly.
• This tool empowers non-specialists in quantitative microscopy.