Automated segmentation and quantitative analysis of organelle morphology, localization and content using CellProfiler
Sebastiaan N J Laan1,2, Richard J Dirven1, Petra E Bürgisser2
1Internal Medicine, Division of Thrombosis and Hemostasis, Leiden University Medical Center, Leiden, The Netherlands.
Plos One
|June 14, 2023
Summary
Automated CellProfiler pipelines, OrganelleProfiler and OrganelleContentProfiler, enable unbiased quantification of secretory organelles like Weibel-Palade bodies (WPBs) in cells. These tools offer high-throughput analysis for diverse cell and organelle types.
Area of Science:
- Cell Biology
- Microscopy and Imaging
- Bioinformatics
Background:
- Confocal microscopy is crucial for studying secretory organelles, but cell heterogeneity necessitates automated analysis.
- Quantifying organelle number, size, shape, and location requires unbiased, high-throughput methods.
Purpose of the Study:
- To develop and validate automated CellProfiler pipelines for quantitative analysis of secretory organelles.
- To assess the utility of these pipelines in diverse cell types and for specific biological questions.
Main Methods:
- Development of two CellProfiler pipelines: OrganelleProfiler and OrganelleContentProfiler.
- Application of pipelines to confocal images of endothelial colony forming cells (ECFCs) and human embryonic kidney 293T (HEK293T) cells.
- Validation of pipeline measurements using Fiji software.
Main Results:
- Pipelines accurately quantify cell and organelle parameters (count, size, shape, localization).
- Demonstrated ability to measure changes in Weibel-Palade body (WPB) size and localization in response to cellular stimuli.
- Successful quantification of protein localization (Rab27A) within or on organelles.
Conclusions:
- The developed pipelines offer a powerful, automated tool for high-processing quantitative characterization of diverse cell and organelle types.
- These freely available and editable pipelines significantly advance organelle analysis in cell biology research.
