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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: Jun 7, 2026

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Published on: July 25, 2022

Dispersion-based pulse shaping for multiplexed two-photon fluorescence microscopy.

Guillaume Labroille¹, Rajesh S Pillai, Xavier Solinas

¹Laboratoire d’Optique et Biosciences, Ecole Polytechnique, CNRS, and INSERM U696, 91128 Palaiseau, France.

|October 23, 2010

Summary

This summary is machine-generated.

We developed a simple method for selective two-photon excited fluorescence microscopy using shaped optical pulses. This technique enhances microscopy performance and simplifies experimental setups for clearer imaging in biological samples.

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Transmission of Multiple Signals through an Optical Fiber Using Wavefront Shaping

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Published on: March 20, 2017

Area of Science:

Biomedical Optics
Microscopy
Fluorescence Imaging

Background:

Two-photon excited fluorescence microscopy (TPEFM) offers enhanced imaging depth and reduced phototoxicity compared to single-photon techniques.
Achieving selective excitation in TPEFM is crucial for high-resolution imaging and minimizing photodamage.
Current methods for pulse shaping can be complex and limit experimental flexibility.

Purpose of the Study:

To demonstrate a novel, simplified scheme for generating shaped optical pulses for selective TPEFM.
To improve the performance and efficiency of TPEFM using broadband shaped pulses.
To enable simultaneous, selective imaging in biological specimens.

Main Methods:

Utilized a broadband oscillator to generate a pulse train.
Split the pulse train into two sub-pulses, introducing differential dispersion via varying glass path lengths.
Recombined the sub-pulses to achieve pulse-shape switching at 150 MHz.
Employed time-resolved photon counting detection for simultaneous image acquisition.

Main Results:

Successfully demonstrated selective two-photon excitation using the dispersive pulse-shaping scheme.
Obtained two simultaneous images with selective excitation from a single experimental setup.
Validated the technique in imaging a live embryo, showcasing its biological applicability.
Achieved improved performance in selective microscopy compared to previous broadband pulse-shaping methods.

Conclusions:

The developed dispersive optical component scheme provides an efficient and simplified method for selective TPEFM.
This approach enhances the performance of selective microscopy with broadband shaped pulses.
The technique offers a practical advancement for biological imaging applications requiring precise excitation control.