How To Characterize Individual Nanosize Liposomes with Simple Self-Calibrating Fluorescence Microscopy
Kim I Mortensen1, Chiara Tassone1, Nicky Ehrlich1
1Department of Micro- and Nanotechnology , Technical University of Denmark , Kongens Lyngby , DK-2800 , Denmark.
Nano Letters
|April 4, 2018
Summary
Characterizing nanosize lipid vesicles is crucial for nanotechnology and biology applications. This study introduces a dual-color fluorescence method to individually assess vesicle properties, revealing that only a fraction meet ideal structural and encapsulation standards.
Area of Science:
- Nanotechnology and biology interface
- Lipid vesicle characterization
- Biophysical analysis
Background:
- Nanosize lipid vesicles are vital for drug delivery and chemical reactions.
- Vesicle samples typically exhibit heterogeneity in size and structure.
- Individual vesicle characterization is essential for accurate experimental interpretation.
Purpose of the Study:
- To develop a method for individual characterization of nanosize lipid vesicles.
- To identify ideal vesicles based on size, shape, lamellarity, and encapsulation efficiency.
- To enable accurate calibration of dual-color fluorescence microscopy images.
Main Methods:
- Dual-color fluorescence labeling of lipid bilayers and lumens.
- Image analysis of vesicle spots to determine intensity and width.
- Calibration of microscopy images using ideal vesicles identified in situ.
Main Results:
- A method was established to yield size, shape, lamellarity, and encapsulation efficiency for each vesicle.
- Ideal vesicles were identified based on specific imaging data relationships.
- Non-ideal vesicles were characterized by deviations from the calibrated relationships.
Conclusions:
- The developed method allows for precise characterization of individual nanosize lipid vesicles.
- Contrary to assumptions, only a fraction of extruded vesicles are structurally ideal.
- In situ calibration using ideal vesicles enhances microscopy accuracy and reduces error.
Keywords:
Liposomesdual-color fluorescence microscopyencapsulation efficiencylamellaritysingle-particle analysisvesiclesMore Related Videos
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