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Updated: May 14, 2026
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Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes
Published on: August 24, 2021
Automated podosome identification and characterization in fluorescence microscopy images
Marjolein B M Meddens1, Bernd Rieger, Carl G Figdor
1Department of Tumor Immunology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.
Summary
A new algorithm accurately identifies podosome cores, crucial cellular adhesion structures, by analyzing F-actin. This tool aids in understanding podosome function and protein distribution in cellular invasion.
Area of Science:
- Cell Biology
- Biophysics
- Biochemistry
Background:
- Podosomes are actin-rich adhesion structures critical for cell invasion and matrix degradation.
- Identifying podosomes is challenging due to the presence of actin in other cellular structures.
- Automated segmentation of podosomes requires distinguishing them from non-podosomal F-actin networks.
Purpose of the Study:
- To develop a quantitative image analysis algorithm for accurate podosome core segmentation.
- To validate the algorithm's performance against expert analysis and other methods.
- To apply the algorithm for studying the effects of cytoskeletal inhibitors and protein distribution.
Main Methods:
- Developed a quantitative image analysis algorithm using sequential local and global thresholding.
- Validated algorithm performance by comparing its podosome identification and quantification with expert analysis.
- Applied the algorithm to analyze F-actin intensity under actin polymerization and myosin II inhibition.
- Expanded core segmentations to investigate cytoskeletal adaptor protein distribution.
Main Results:
- The algorithm successfully identifies up to 76% of podosome cores, excluding other F-actin structures.
- Algorithm performance in podosome identification and quantification is comparable to expert analysis.
- The algorithm revealed effects of actin polymerization and myosin II inhibition on actin intensity.
- Differential distribution of cytoskeletal adaptor proteins within the podosome ring was observed.
Conclusions:
- The developed algorithm provides a valuable tool for rapid and accurate large-scale analysis of podosomes.
- This method enhances the understanding of podosome structure, function, and regulation.
- The algorithm facilitates research into cellular adhesion, matrix degradation, and invasion processes.
