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The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...

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Related Experiment Video

Updated: Jan 10, 2026

Visualization of Organelles In Situ by Cryo-STEM Tomography

Visualization of Organelles In Situ by Cryo-STEM Tomography

Published on: June 23, 2023

Visualizing intraorganellar ultrastructures, dynamics, and interactions with open-access background-free Lock-in-SIM.

Wenjie Liu^1,2, Meng Zhang³, Wenbin Zhu⁴

¹Department of Biochemistry, University of Oxford, Oxford, UK. wenjie.liu@chem.ox.ac.uk.

Nature Communications

|November 28, 2025

Summary

This summary is machine-generated.

Lock-in-SIM, a new 2D super-resolution microscopy technique, overcomes background noise and blur to reveal detailed live-cell organelle structures. This advanced imaging method enhances understanding of mitochondrial and ER-lysosome dynamics.

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Area of Science:

Cell Biology
Microscopy
Biophysics

Background:

Structured Illumination Microscopy (SIM) offers fast, gentle live-cell super-resolution imaging.
SIM is limited by reconstruction artifacts from out-of-focus blur and background.
Analyzing densely packed intraorganellar ultrastructures is challenging due to SIM's spatial resolution limits.

Purpose of the Study:

To develop an advanced SIM framework to overcome existing limitations.
To improve the analysis of challenging intraorganellar ultrastructures in live cells.
To enhance data fidelity and quantifiability in super-resolution microscopy.

Main Methods:

Developed Lock-in-SIM, an open-access 2D SIM framework.
Leveraged intrinsic modulation differences of volumetric sample structures.
Implemented background elimination and maximized high-frequency extraction for improved resolution.

Main Results:

Lock-in-SIM enables efficient optical sectioning and extends imaging depth.
Demonstrated superior visualization of challenging live-cell intraorganellar ultrastructures.
Achieved enhanced data fidelity and quantifiability compared to conventional SIM.

Conclusions:

Lock-in-SIM significantly improves super-resolution imaging of complex cellular structures.
The technique provides new insights into mitochondrial fission and ER-lysosome interactions.
Advanced SIM imaging aids in understanding organelle structural remodeling mechanisms.