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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.

Confocal Fluorescence Microscopy

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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: Jul 9, 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

Multifocal multiphoton microscopy.

J Bewersdorf, R Pick, S W Hell

|December 19, 2007

Summary

This summary is machine-generated.

This study introduces a novel multiphoton fluorescence microscope for high-resolution 3D imaging. The advanced system achieves significantly faster imaging speeds, enabling real-time observation of live samples.

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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:

Biophysics
Microscopy
Optical Engineering

Background:

Multiphoton microscopy offers optical sectioning and deep tissue penetration.
High-resolution imaging of dynamic biological processes requires fast acquisition speeds.
Current multiphoton systems can be limited by speed or complexity.

Purpose of the Study:

To develop a real-time, direct-view multiphoton excitation fluorescence microscope.
To achieve high-resolution three-dimensional imaging with enhanced efficiency.
To enable direct visual observation of high-resolution images of live samples.

Main Methods:

Utilized a rotating microlens disk to split laser light into multiple beams.
Employed a mode-locked titanium-sapphire laser for near-infrared excitation.
Integrated a high-speed camera for video-rate image readout.

Main Results:

Achieved axial resolutions of 0.84 µm (oil) and 1.4 µm (water) at 780 nm excitation.
Demonstrated imaging speeds of 225 frames per second.
Showcased a 40-100 fold increase in imaging efficiency compared to single-beam systems.
Enabled direct, real-time, high-resolution visualization of two-photon images.

Conclusions:

The developed multiphoton microscope provides a significant advancement in imaging speed and efficiency.
This technology facilitates high-resolution, real-time observation of biological samples.
The system offers a direct-view capability, enhancing usability for researchers.