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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 developed.

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Updated: May 26, 2026

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Multicolor super-resolution fluorescence imaging via multi-parameter fluorophore detection.

Mark Bates¹, Graham T Dempsey, Kok Hao Chen

¹School of Engineering and Applied Sciences, Graduate program in Biophysics, Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts, 02138, USA.

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry

|January 4, 2012

Summary

This summary is machine-generated.

Researchers developed multicolor super-resolution microscopy using multi-parameter detection to image six cellular targets simultaneously. This advanced technique improves fluorophore discrimination for enhanced visualization of complex cellular environments at nanometer resolution.

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Last Updated: May 26, 2026

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

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Published on: February 24, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

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Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Area of Science:

Cellular Biology
Microscopy
Biophysics

Background:

Simultaneous imaging of multiple cellular components with nanometer resolution is crucial for understanding cellular complexity.
Current multicolor super-resolution microscopy faces limitations in the number of simultaneously imaged targets and color channel crosstalk.

Purpose of the Study:

To demonstrate a multicolor stochastic optical reconstruction microscopy (STORM) method utilizing multi-parameter detection for improved fluorophore discrimination.
To overcome limitations in simultaneous imaging and crosstalk in multicolor super-resolution fluorescence microscopy.

Main Methods:

Developed a multi-parameter detection strategy for stochastic optical reconstruction microscopy (STORM).
Utilized fluorescence activation wavelength and emission color to differentiate photo-activatable fluorescent probes.
Employed Alexa 750 (near-infrared) and Alexa 647 (red) cyanine dyes, along with photo-activation enhancing fluorophores.

Main Results:

Achieved two-color super-resolution imaging with Alexa 750 and Alexa 647, quantifying crosstalk and registration accuracy.
Enabled multi-parameter detection of six distinct probes through combinatorial dye pairing.
Acquired six-color super-resolution fluorescence images of a model sample.

Conclusions:

The multi-parameter detection strategy significantly enhances fluorophore discrimination in multicolor STORM.
This approach substantially improves the ability to visualize multiple cellular targets with sub-diffraction-limit resolution.
Promises to advance the understanding of complex cellular environments through high-resolution multicolor imaging.