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Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
Published on: September 5, 2019
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Spatial heterogeneity of the cytosol revealed by machine learning-based 3D particle tracking
Grace A McLaughlin1, Erin M Langdon1, John M Crutchley1
1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
Molecular Biology of the Cell
|May 14, 2020
Summary
Cytosol physical properties vary significantly in space and time. Genetically encoded multimeric nanoparticles (GEMs) reveal localized zones of low diffusivity within fungal cells, highlighting cellular heterogeneity.
Area of Science:
- Cell Biology
- Biophysics
Background:
- Cytosol spatial structure and physical properties remain poorly understood.
- Measuring cytosol material state is difficult due to heterogeneity.
- Genetically encoded multimeric nanoparticles (GEMs) enable studying cytosol at 20-60 nm scales.
Purpose of the Study:
- To develop and apply an image analysis pipeline for 3D imaging of GEMs in fungi.
- To quantitatively visualize spatially varying cytosol diffusivity.
- To investigate cytosol heterogeneity in multinucleate fungi.
Main Methods:
- Developed a 3D image analysis pipeline for GEMs.
- Utilized a neural network for 3D particle tracking.
- Created quantitative visualizations of diffusivity patterns.
Main Results:
- Detected substantial variability in cytosol properties.
- Identified zones of low GEM diffusivity at hyphal tips and near nuclei.
- Observed significant cell-to-cell variability in average GEM diffusivity.
Conclusions:
- Cytosol physical properties vary significantly in space and time.
- Spatial variations in cytosol diffusivity contribute to cellular heterogeneity.
- Cell-to-cell variability in diffusivity impacts populations.
