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Published on: June 21, 2017
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[Improvement and application of a fast labeling system for bulk internalized vesicles]
Ye Rong1, Zhong-Yan DU1, Gui-Feng Xiao2
1Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 311402, China.
Sheng Li Xue Bao : [Acta Physiologica Sinica]
|June 22, 2018
Summary
Researchers developed a fast, convenient assay to track bulk internalized vesicles like macropinosomes and lysosomes in live cells. This method uses dye injection and live imaging, avoiding dye washout for efficient vesicle trafficking studies.
Area of Science:
- Cell biology
- Molecular biology
- Live-cell imaging
Background:
- Studying the trafficking of internalized vesicles, such as macropinosomes and lysosomes, is crucial for understanding cellular processes.
- Existing methods for vesicle tracking can be cumbersome, involving multiple steps like dye washout and cell transfer.
Purpose of the Study:
- To establish an efficient and convenient assay for studying the trafficking of bulk internalized vesicles in live cells.
- To develop a labeling system that is fast and avoids the need for dye washout or cell transfer.
Main Methods:
- Adapted the axon turning assay for vesicle trafficking studies.
- Injected indicator or fluorescent dyes via micropipette with air pressure into cell cultures to create a stable gradient.
- Utilized live-cell imaging to record the entire vesicle trafficking process.
Main Results:
- Successfully established an efficient and convenient assay for live-cell vesicle trafficking.
- Demonstrated a fast labeling system for bulk internalized vesicles (e.g., macropinosomes, lysosomes).
- The assay is compatible with live-cell imaging systems and eliminates the need for dye washout and cell transfer.
Conclusions:
- The developed assay provides an effective and rapid method for labeling and tracking bulk internalized vesicles.
- This technique simplifies the study of vesicle trafficking in live cells.
- The assay can be readily integrated with existing live-cell imaging platforms for enhanced research capabilities.
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