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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: May 3, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Fixed pattern noise in localization microscopy.

Patrick Fox-Roberts¹, Tianqi Wen, Klaus Suhling

¹Randall Division of Cell and Molecular Biophysics, New Hunt's House, Guy's Campus, King's College London, SE1 1UL (United Kingdom).

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry

|February 1, 2014

Summary

This summary is machine-generated.

Fixed pattern noise in localization microscopy can affect emitter position accuracy. Different fitting algorithms and pixel sizes impact this noise, leading to 2-4 nm accuracy variations, crucial for high-precision applications.

Keywords:

algorithms fixed pattern noise fluorescence localization microscopy super-resolution

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Area of Science:

Biophysics
Optical Microscopy
Nanotechnology

Background:

Localization microscopy enhances fluorescence microscopy resolution by fitting individual fluorophore positions.
Accurate emitter localization is critical for super-resolution imaging.

Purpose of the Study:

To investigate fixed pattern noise in localization microscopy reconstructions.
To evaluate the impact of different fitting algorithms and pixel sizes on localization accuracy.

Main Methods:

Analysis of reconstructions from various fitting algorithms.
Simulation of fixed pattern noise under different conditions.
Assessment of noise impact based on pixel size relative to the point spread function.

Main Results:

Fixed pattern noise can occur during sub-pixel fitting, even if not immediately apparent.
The degree of fixed pattern noise varies significantly with the fitting algorithm used.
Pixel size relative to the point spread function influences the extent of fixed pattern noise.

Conclusions:

Fixed pattern noise introduces systematic errors in emitter position estimation.
Accuracy variations of 2-4 nm were observed for pixel sizes of 80-170 nm.
These variations are non-negligible for applications requiring very high precision in localization microscopy.