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Measuring Intracellular Vesicle Density and Dispersion Using Fluorescence Microscopy and ImageJ/FIJI
Natália Fernanda do Couto1, Thamires Queiroz-Oliveira2, Maria Fátima Horta3
1Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil.
Bio-Protocol
|March 4, 2021
Summary
This study introduces a simple, cost-effective method to analyze intracellular vesicle distribution using immunofluorescence and ImageJ/FIJI software. This technique aids in understanding cellular processes like secretion and membrane repair by quantifying vesicle density and dispersion.
Area of Science:
- Cell Biology
- Molecular Biology
- Biophysics
Background:
- Intracellular vesicles are crucial for cell signalling, secretion, and plasma membrane repair.
- Their strategic distribution is vital for various cellular functions within endocytic and secretory pathways.
Purpose of the Study:
- To present a simple, affordable, and efficient tool for analyzing intracellular vesicle distribution.
- To enable quantification of vesicle density and dispersion under diverse experimental conditions.
Main Methods:
- Combining immunofluorescence with pixel-based quantification software (ImageJ/FIJI).
- Tracking fluorescent vesicles based on their position relative to cell nuclei.
- Utilizing open-source, free software for broad accessibility.
Main Results:
- The method allows for quantitative analysis of intracellular vesicle distribution.
- It provides insights into vesicle density and dispersion patterns.
- Demonstrates a practical approach for studying lysosomes, endosomes, Golgi vesicles, and secretory granules.
Conclusions:
- The presented method is an accessible and efficient tool for analyzing intracellular vesicle distribution.
- It has wide applicability within the scientific community for studying cellular processes.
- Facilitates a deeper understanding of the role of vesicle localization in cell function.
