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Correlative Super-resolution and Electron Microscopy to Resolve Protein Localization in Zebrafish Retina
Published on: November 10, 2017
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Comprehensive correlation analysis for super-resolution dynamic fingerprinting of cellular compartments using the
L Scipioni1, L Lanzanó2, A Diaspro2,3
1Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, 92697, CA, USA.
Nature Communications
|December 4, 2018
Summary
A new super-resolution concept using the Airyscan detector enables advanced fluctuation correlation spectroscopy. This method distinguishes nanoscale cellular environments by analyzing molecular fluctuations of free GFP in live cells.
Area of Science:
- Biophysics
- Cell Biology
- Microscopy
Background:
- Super-resolution microscopy offers enhanced spatial resolution.
- Fluctuation correlation spectroscopy (FCS) analyzes molecular dynamics.
- Traditional FCS has limitations in speed and spatial coverage.
Purpose of the Study:
- To introduce a novel fluctuation correlation spectroscopy (FCS) concept utilizing the Airyscan detector.
- To leverage super-resolution capabilities for enhanced molecular analysis.
- To demonstrate the ability to differentiate subcellular environments using this technique.
Main Methods:
- Utilized the Zeiss LSM 880 with its Airyscan detector (32 hexagonal array detectors).
- Applied time and spatial correlation methods to fluctuation data.
- Acquired millions of frames rapidly for robust statistical analysis.
- Analyzed molecular fluctuations of free GFP diffusing in live cells.
Main Results:
- The Airyscan detector enables simultaneous multi-point time and spatial correlation analysis.
- High frame rates provide statistically robust fluctuation data.
- Comprehensive fluctuation analysis successfully distinguished different subcellular environments at the nanoscale.
Conclusions:
- The developed super-resolution FCS concept is a powerful tool for nanoscale analysis.
- This method allows for the characterization of distinct cellular microenvironments.
- Airyscan-based FCS provides new insights into molecular dynamics within live cells.
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