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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

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: Dec 5, 2025

Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus

Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus

Published on: November 6, 2018

Photobleaching reduction in modulated super-resolution microscopy.

Jafar H Ghithan¹, Jennifer M Noel², Thomas J Roussel³

¹University of Louisville, Department of Physics and Astronomy, 215 Eastern Pkwy, Louisville, Kentucky, United States, 40292.

Microscopy (Oxford, England)

|October 16, 2020

Summary

This summary is machine-generated.

This study introduces a modulated excitation approach for Stimulated Emission Depletion (STED) microscopy, significantly reducing laser power and photobleaching. This advancement enables clearer super-resolution imaging, especially for sensitive biological samples.

Keywords:

STED microscopy fluorescence nanoscopy imaging laser photobleaching super-resolution

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Last Updated: Dec 5, 2025

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Ground State Depletion Super-resolution Imaging in Mammalian Cells

Ground State Depletion Super-resolution Imaging in Mammalian Cells

Published on: November 5, 2017

Area of Science:

Microscopy
Optical Imaging
Biophysics

Background:

Stimulated Emission Depletion (STED) microscopy has advanced far-field imaging.
Continuous wave (CW) STED microscopy often requires high laser power, leading to photobleaching.
Photobleaching limits imaging quality and sample viability in STED.

Purpose of the Study:

To reduce photobleaching and optical power requirements in STED microscopy.
To improve the quality of super-resolution images obtained with STED.
To develop a more robust STED technique for biological samples.

Main Methods:

Implemented a synchronous detection approach with a modulated excitation laser.
Maintained a continuous wave (CW) depletion laser.
Utilized a lock-in amplifier for frequency filtering of the fluorescence signal.

Main Results:

Achieved significant reduction in optical power for both excitation and depletion lasers.
Observed measurable decreases in photobleaching effects.
Acquired high-quality super-resolution images with reduced scattering and background noise.

Conclusions:

The modulated STED approach effectively lowers laser power and minimizes photobleaching.
This technique enhances STED imaging capabilities for delicate biological specimens.
Reduced power requirements make STED microscopy more accessible for various applications.