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Related Experiment Video

Updated: Sep 30, 2025

Three-Dimensional Imaging of Tumor-Bearing Tissue Using the Iterative Bleaching Extends Multiplexity Approach

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Published on: April 25, 2025

Lifetime based axial contrast enable simple 3D-STED imaging.

Yuanqing Ma^1,2,3, Alex Macmillan⁴, Ying Yang⁵

¹EMBL Australia Node in Single Molecule Science, and ARC Centre of Excellence in Advanced Molecular Imaging School of Medical Sciences, The University of New South Wales, 2052 Sydney, Australia.

Methods and Applications in Fluorescence

|March 15, 2022

Summary

This summary is machine-generated.

This study introduces a novel method to enhance axial resolution in Stimulated Emission Depletion (STED) microscopy using a lifetime-based contrast generated by ITO-coated coverslips. This technique improves 3D imaging without requiring microscope modifications.

Keywords:

3D-STED Lifetime contrast MIET

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Three-Dimensional Imaging of Aortic Tissues in Atherosclerosis

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Area of Science:

Super-resolution microscopy
Biophysics
Optical imaging

Background:

Stimulated Emission Depletion (STED) microscopy enhances lateral resolution but struggles with axial resolution.
Achieving isotropic 3D STED resolution requires precise optical alignment for axial interference patterns.
Current methods for improving axial resolution in STED are often complex and optically demanding.

Purpose of the Study:

To develop a straightforward method for improving axial resolution in STED microscopy.
To introduce a lifetime-based axial contrast mechanism for 3D STED imaging.
To demonstrate the application of this technique in biological samples without optical modifications.

Main Methods:

Utilized an Indium Tin Oxide (ITO) coated glass coverslip for STED imaging.
Leveraged surface plasmon resonance and Metal-Induced Energy Transfer (MIET) effect on the ITO surface.
Generated an axial lifetime gradient extending over 400 nm from the ITO surface.

Main Results:

Achieved a ∼20% dynamic range in the lifetime gradient.
Successfully performed 3D STED imaging of tubulin fibers in COS-7 cells.
Revealed the vertical displacement of individual tubulin fibers with enhanced axial contrast.

Conclusions:

The proposed lifetime-based axial contrast method offers a simple yet effective approach to improve 3D STED resolution.
This technique requires no optical modifications to standard STED microscopes equipped with pulsed lasers.
The method provides a new avenue for high-resolution 3D imaging in biological and material sciences.