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Super-STORM: Molecular Modeling to Achieve Single-molecule Localization with STORM Microscopy
Zhichao Fan1,2, Zbigniew Mikulski3, Sara McArdle3
1Division of Inflammation Biology, La Jolla Institute for Immunology, 9420 Athena Circle Drive, La Jolla, CA 92037, USA.
STAR Protocols
|October 28, 2020
Summary
SuperSTORM combines stochastic optical reconstruction microscopy (STORM) with molecular modeling for enhanced cell imaging. This technique improves the resolution and analysis of cell-surface molecules, including neutrophil integrins.
Area of Science:
- Biophysics
- Cell Biology
- Microscopy
Background:
- Stochastic optical reconstruction microscopy (STORM) enables super-resolution imaging.
- Analyzing molecular distribution and orientation on cell surfaces remains challenging.
Purpose of the Study:
- Introduce the SuperSTORM technique, integrating STORM with molecular modeling.
- Optimize STORM imaging and analysis for cell-surface molecules.
Main Methods:
- Combined stochastic optical reconstruction microscopy (STORM) and molecular modeling.
- Developed methods for image processing, including blink elimination and colocalization analysis.
- Adapted STORM imaging for cells within microfluidic systems.
Main Results:
- SuperSTORM enhances the resolution of STORM imaging by utilizing biomolecule-inherent parameters.
- The technique accurately identifies molecular clusters, orientations, and distributions.
- Successfully applied to neutrophil integrins and adaptable for other cell-surface molecules.
Conclusions:
- SuperSTORM offers a powerful approach for high-resolution imaging and analysis of cell-surface molecules.
- The protocol extends STORM capabilities to microfluidic cell studies.
- Provides a framework for detailed molecular investigation on cell surfaces.
