Automated noninvasive epithelial cell counting in phase contrast microscopy images with automated parameter selection

Area of Science:

• Cell Biology
• Microscopy Techniques
• Quantitative Imaging

Background:

• Traditional cell counting for proliferation studies is often destructive, preventing longitudinal monitoring of cell growth.
• Current methods are time-consuming and resource-intensive, particularly for adherent cell cultures.
• There is a need for non-destructive, efficient cell counting techniques in cell culture research.

Purpose of the Study:

• To develop an inexpensive, rapid, and widely applicable phase contrast microscopy-based technique for non-destructive cell counting.
• To enable longitudinal tracking of cell growth in cultures.
• To provide a validated method for generating cell growth curves.

Main Methods:

• A phase contrast microscopy technique was developed, emulating contrast changes from defocused bright field images.
• Image processing involved global thresholding for segmentation, with cell counts derived from segmented regions.
• Linear discriminant analysis was employed to remove spurious noise and improve cell count accuracy.

Main Results:

• The developed technique provides accurate cell counts, validated against manual counting.
• The method successfully generated growth curves for oral keratinocyte cultures.
• The technique demonstrated broad applicability for researchers with standard laboratory imaging equipment.

Conclusions:

• A novel, cost-effective, and rapid phase contrast microscopy method for non-destructive cell counting has been established.
• This technique overcomes the limitations of destructive cell counting, allowing for longitudinal studies.
• The method is suitable for generating reliable cell growth curves and has wide utility in cell culture research.