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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.

Total Internal Reflection Fluorescence Microscopy

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.

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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Updated: Jun 7, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors

Published on: May 30, 2016

Full-wavelet approach for fluorescence diffuse optical tomography with structured illumination.

Nicolas Ducros¹, Cosimo D'andrea, Gianluca Valentini

¹Instituto di Fotonica e Nanotecnologie (IFN-CNR)-Dipartimento di Fisica, Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 Milan, Italy. nicolas.ducros@polimi.it

|November 3, 2010

Summary

This summary is machine-generated.

We developed a faster fluorescence optical tomography method using wavelet transforms for quicker imaging. This technique significantly reduces scan and computation times, making it ideal for real-time in vivo applications.

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Area of Science:

Biomedical optics
Medical imaging
Fluorescence imaging

Background:

Fluorescence optical tomography (FOT) is a powerful imaging modality.
Traditional FOT reconstruction methods can be time-consuming.
In vivo applications require rapid imaging techniques.

Purpose of the Study:

To develop a fast reconstruction method for fluorescence optical tomography.
To reduce acquisition and computation times in FOT.
To enable efficient in vivo FOT imaging.

Main Methods:

Utilized structured illumination with wavelet-patterned illuminations.
Applied wavelet transform to acquired measurements.
Validated the method using experimental data.

Main Results:

Achieved significant reduction in acquisition time compared to classical scanning.
Demonstrated substantial decrease in computation time.
Successfully validated the method on experimental data.

Conclusions:

The proposed wavelet-based FOT reconstruction is significantly faster.
This method offers a viable solution for time-constrained in vivo imaging.
The approach holds promise for advancing biomedical optical imaging.