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Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins

Automated Quantification and Analysis of Cell Counting Procedures Using ImageJ Plugins

Published on: November 17, 2016

Automated cell colony counting and analysis using the circular Hough image transform algorithm (CHiTA).

J M Bewes¹, N Suchowerska, D R McKenzie

¹School of Physics, University of Sydney, Sydney, NSW, Australia. jbewes@physics.usyd.edu.au

Physics in Medicine and Biology

|October 7, 2008

Summary

This study introduces an automated cell colony counting method using an unapproximated Hough transform for robust clonogenic analysis. The flexible system offers deeper insights than manual or existing automated techniques.

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Area of Science:

Cell Biology
Image Analysis
Computational Biology

Background:

Clonogenic assays are crucial for evaluating cell survival and proliferation.
Manual colony counting is labor-intensive and prone to errors.
Existing automated methods lack depth in clonogenic analysis.

Purpose of the Study:

To develop a flexible and robust automated method for cell colony counting.
To provide more in-depth clonogenic analysis compared to current approaches.
To enable the use of standard desktop scanners for image acquisition.

Main Methods:

Implemented the full, unapproximated Hough transform for colony identification.
Utilized image pre-processing techniques: erosion, dilation, and Gaussian smoothing.

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Employed intensity gradient field discrimination for colony edge detection.

Main Results:

The method accurately counts colonies, resolves merged colonies, and identifies those near flask edges.
Evaluated additional parameters: regional counts, average area, nearest neighbor distances, and radial distribution.
Achieved high accuracy validated by manual counting and a novel Monte Carlo simulation.

Conclusions:

The developed automated system enhances the utility of clonogenic assays by providing spatial and qualitative data.
This approach allows for the analysis of spatially-variant cytotoxic effects.
The method is adaptable to different cell lines, flask types, and plating densities, requiring no specialized hardware.