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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...

Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: Mar 8, 2026

Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment

Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment

Published on: January 6, 2026

Simultaneous multiplane imaging-based localization encoded (SMILE) microscopy for super-resolution volume imaging.

Partha Pratim Mondal¹

¹Nanobioimaging Laboratory, Department of Instrumentation and Applied Physics, Indian Institute of Science, Bangalore, 560012, India.

Microscopy Research and Technique

|January 21, 2017

Summary

This summary is machine-generated.

We introduce a widefield super-resolution microscopy technique for rapid 3D imaging. This method precisely localizes single molecules, enabling fast, high-resolution volume reconstruction without extensive scanning.

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Last Updated: Mar 8, 2026

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Area of Science:

Microscopy
Optical Imaging
Biophysics

Background:

Conventional super-resolution microscopy often requires slow scanning, limiting 3D imaging speed.
Achieving high-resolution volumetric data necessitates precise localization of fluorescent signals in three dimensions.

Discussion:

The proposed technique encodes single molecules to their respective planes for accurate localization.
System point spread function (PSF) calibration using sub-diffraction beads is crucial for distinguishing focal and off-focal planes.
Multiple cut-offs applied to PSFs enable identification and sorting of single molecules for 3D reconstruction.

Key Insights:

This widefield-based method achieves rapid super-resolution volume imaging.
It eliminates the need for laborious z-plane scanning, significantly increasing imaging speed.
The technique minimizes radiation dose to the specimen, preserving biological integrity.

Outlook:

Potential applications in live-cell imaging and dynamic biological process visualization.
Further optimization could enhance resolution and imaging speed for complex biological systems.
Adaptation for different types of super-resolution microscopy techniques is feasible.