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Atomic Force Microscopy

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Related Experiment Video

Updated: Sep 10, 2025

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

Published on: July 20, 2019

Imaging Nuclear Envelopes Using Correlative AFM/Fluorescence Microscopy.

Emilie Costes¹, Anthony Vial², Christine Doucet³

¹Centre de Biologie Structurale, INSERM, CNRS, University of Montpellier, Montpellier, France.

Methods in Molecular Biology (Clifton, N.J.)

|August 20, 2025

Summary

This summary is machine-generated.

This study presents a new method for preparing nuclear envelopes (NEs) from mammalian cells, preserving nuclear pore complex (NPC) structure. This technique allows for high-resolution imaging of both inner and outer nuclear membranes using atomic force microscopy (AFM).

Keywords:

Atomic force microscopy (AFM)Direct stochastic optical reconstruction microscopy (dSTORM)Fluorescence microscopy Mammalian cells Nuclear envelope Nuclear pore complex (NPC)Single-molecule localization microscopy (SMLM)

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Single-Molecule Imaging of Nuclear Transport

Single-Molecule Imaging of Nuclear Transport

Published on: June 9, 2010

Area of Science:

Cell Biology
Biophysics
Microscopy

Background:

The nuclear envelope (NE) is a crucial eukaryotic structure with inner and outer membranes.
Existing methods for NE preparation can alter nuclear pore complex (NPC) morphology due to membrane tension loss.
Atomic force microscopy (AFM) is a powerful tool for imaging NE structures.

Purpose of the Study:

To develop a novel method for preparing intact nuclear envelopes (NEs) from cultured mammalian cells.
To preserve the native structure of nuclear pore complexes (NPCs) during sample preparation.
To enable high-resolution imaging of both inner and outer nuclear membranes using AFM and correlative microscopy.

Main Methods:

Extraction of nuclei from cultured mammalian cells and affixing them to glass substrates.
Gentle opening of nuclei to maintain membrane tension and NPC integrity.
Immuno-labeling combined with AFM and direct Stochastic Optical Reconstruction Microscopy (dSTORM) for correlative imaging.

Main Results:

The novel preparation method successfully preserves NE membrane tension and NPC structure.
High-resolution AFM imaging of both inner and outer nuclear membranes is achieved.
Correlative AFM/dSTORM imaging provides detailed structural and molecular information.

Conclusions:

This protocol offers a robust approach for studying NEs and NPCs with preserved morphology.
The method is adaptable for various cellular models, including plant cells and tissues.
It facilitates advanced correlative imaging techniques for deeper biological insights.