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Related Experiment Video

Updated: Jul 25, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Extracting nanoscale membrane morphology from single-molecule localizations.

Zach Marin¹, Lukas A Fuentes², Joerg Bewersdorf³

¹Auckland Bioengineering Institute, University of Auckland, Auckland, New Zealand; Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut; Department of Biomedical Engineering, Yale University, New Haven, Connecticut.

Biophysical Journal

|June 25, 2023

Summary

This summary is machine-generated.

NanoWrap reconstructs nanoscale membrane surfaces from generalized single-molecule localization microscopy data, enabling cellular structure studies. This method overcomes limitations of electron and fluorescence microscopy for visualizing subcellular shape changes.

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Last Updated: Jul 25, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

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Published on: June 30, 2018

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Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

Area of Science:

Cell Biology
Biophysics
Microscopy

Background:

Nanoscale membrane surface reconstruction is crucial for understanding subcellular dynamics.
Electron microscopy has limitations in extracting surfaces from specific labels.
Current fluorescence microscopy methods offer poor resolution or require extensive manual data curation.

Purpose of the Study:

To develop a novel method for extracting nanoscale membrane surfaces from generalized single-molecule localization microscopy (SMLM) data.
To enable the study of shapes of specifically labeled membranous structures within cells.
To overcome the resolution and data quality limitations of existing techniques.

Main Methods:

Development of NanoWrap, a new computational method for surface extraction.
Validation using simulated SMLM data.
Application to experimental SMLM data of cellular structures.

Main Results:

NanoWrap successfully reconstructs nanoscale membrane surfaces from SMLM data.
The method demonstrates robust performance on simulated and experimental datasets.
Reconstructions were achieved for the endoplasmic reticulum and mitochondria.

Conclusions:

NanoWrap provides a powerful new tool for nanoscale membrane surface reconstruction.
The method enhances the study of subcellular structures using SMLM.
NanoWrap is implemented in the open-source Python Microscopy Environment, promoting accessibility.