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Super-Resolution Imaging of Self-Assembled Nanocarriers Using Quantitative Spectroscopic Analysis for Cluster
Janel L Davis1, Yang Zhang1, Sijia Yi1
1Department of Biomedical Engineering, Northwestern University, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
Langmuir : the ACS Journal of Surfaces and Colloids
|February 19, 2020
Summary
Researchers developed a new imaging technique, quantitative spectroscopic analysis for cluster extraction (qSPACE), to precisely visualize self-assembled polymersomes (PS). This method overcomes limitations of traditional microscopy, enabling accurate nanocarrier analysis in native environments.
Area of Science:
- Nanotechnology
- Biophysics
- Microscopy
Background:
- Self-assembled nanocarriers are crucial for bioimaging, diagnostics, and drug delivery.
- Understanding nanocarrier structure-function relationships requires noninvasive visualization in native environments.
- Existing technologies struggle with quantitative nanocarrier visualization, often complicated by fluorescent probe artifacts in single-molecule localization microscopy (SMLM).
Purpose of the Study:
- To develop a method for precise, quantitative visualization of self-assembled polymersomes (PS) in their native environment.
- To overcome challenges posed by nonspecific binding artifacts in SMLM.
- To enable accurate characterization of nanocarriers for improved applications.
Main Methods:
- Spectroscopic point accumulation for imaging in nanoscale topography (sPAINT) was employed to visualize polymersomes with molecular specificity.
- Unique spectral signatures of Nile Red (NR) were analyzed to differentiate specific binding from nonspecific probe adsorption.
- Quantitative spectroscopic analysis for cluster extraction (qSPACE) was developed to enhance localization density and accuracy.
Main Results:
- sPAINT successfully visualized polymersomes with molecular specificity.
- Analysis of NR spectral signatures effectively rejected artifacts from nonspecific binding.
- qSPACE increased localization density fourfold compared to sPAINT, reducing PS size measurement variations to under 5%.
- qSPACE achieved ~20 nm localization precision and a 97% reduction in sample misidentification compared to conventional SMLM.
Conclusions:
- The developed qSPACE method enables highly precise and quantitative imaging of polymersomes.
- This technique significantly reduces artifacts and misidentification, improving nanocarrier characterization.
- qSPACE offers a powerful tool for understanding nanocarrier behavior in biological contexts, advancing their use in various applications.
