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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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Related Experiment Video

Updated: Jun 19, 2026

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

Published on: October 9, 2009

Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

D Baddeley¹, V O Chagin, L Schermelleh

¹Kirchhoff Institut für Physik, University of Heidelberg, Germany.

Nucleic Acids Research

|October 30, 2009

Summary

Super-resolution microscopy revealed a conserved 125 nm size for DNA replication foci, suggesting a basic unit of genome duplication. This technique also identified more replication sites than previously seen.

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Last Updated: Jun 19, 2026

Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

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Published on: October 9, 2009

Super-Resolution Microscopy of the Synaptonemal Complex Within the Caenorhabditis elegans Germline

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Published on: September 13, 2022

Super-resolution Imaging of the Bacterial Division Machinery

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

Area of Science:

Cell Biology
Molecular Biology
Genetics

Background:

DNA replication is a fundamental cellular process occurring within complex macromolecular structures.
Quantitative data is essential for developing and validating models of genome duplication.

Purpose of the Study:

To directly measure and compare the size and number of replication foci in mammalian cells using super-resolution microscopy.
To investigate the potential existence of a basic unit of genome duplication.

Main Methods:

Utilized two super-resolution light microscopy techniques: Spatially Modulated Illumination (SMI) and 3D-Structured Illumination Microscopy (3D-SIM).
Analyzed replication foci size and number in mammalian cells.

Main Results:

Replication foci sizes ranged from 40 nm to 210 nm.
Both SMI and 3D-SIM consistently showed an average focus size of 125 nm, conserved throughout S-phase and independent of labeling.
3D-SIM identified 3- to 5-fold more distinct replication foci compared to previous reports.

Conclusions:

The conserved average size of 125 nm suggests a fundamental unit of genome duplication.
Super-resolution microscopy provides accurate cellular structure measurements comparable to electron microscopy.
High-throughput, multispectral 3D analyses of cellular processes are feasible with these nanoscopy techniques.