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Super-resolution Fluorescence Microscopy

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...

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Updated: Oct 18, 2025

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy iPALM

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Published on: December 1, 2016

Exploring Cell Surface-Nanopillar Interactions with 3D Super-Resolution Microscopy.

Anish R Roy¹, Wei Zhang¹, Zeinab Jahed¹

¹Department of Chemistry, Stanford University, Stanford, California 94305, United States.

|September 28, 2021

Summary

This summary is machine-generated.

Researchers developed a 3D super-resolution microscopy method to precisely map protein distribution on nanoscale membrane structures. This technique reveals how membrane curvature influences protein accumulation, impacting cellular processes like endocytosis.

Keywords:

3D super-resolution fluorescence microscopy cell−material interface endocytosis membrane curvature nanopillars nano−bio interface single-molecule imaging

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Area of Science:

Cellular and Molecular Biophysics
Nanotechnology
Microscopy

Background:

Plasma membrane topography influences cellular processes like endocytosis and actin dynamics.
Existing 2D microscopy lacks resolution to study protein distribution on 100 nm structures.
3D nanostructures (nanopillars) create defined membrane curvatures for studying nano-bio interfaces.

Purpose of the Study:

To develop and validate a 3D super-resolution (SR) microscopy method for nanoscale protein distribution analysis.
To investigate the spatial organization of proteins at the nano-bio interface with high precision (10-20 nm).
To understand how membrane curvature affects protein localization and cellular functions.

Main Methods:

Utilized 3D single-molecule super-resolution localization microscopy.
Employed a silicone-oil immersion objective and 3D double-helix point spread function.
Minimized spherical aberrations between quartz nanopillars and cells for accurate imaging.

Main Results:

Validated the 3D SR method by imaging nanopillar shapes, correlating with electron microscopy.
Observed tight membrane wrapping around nanopillars in cellular reconstructions.
Found cytoplasmic AP-2 protein accumulates with increasing membrane curvature (1/R).
Demonstrated preferential accumulation of AP-2 and actin at positive Gaussian curvature near pillar caps.

Conclusions:

Established a general 3D SR microscopy method for nanoscale protein distribution at the nano-bio interface.
Provided insights into how membrane curvature dictates protein localization, affecting endocytosis.
Highlighted the role of Gaussian curvature in protein recruitment to nanostructured interfaces.