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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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Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

Published on: July 17, 2012

Multiplexed fluorescence mediated tomography with temporal and spectral data.

Ying Mu¹, Vivian Pera¹, Mark Niedre¹

¹Northeastern University, Department of Electrical and Computer Engineering, 360 Huntington Avenue, Boston, Massachusetts 02115, United States.

Journal of Biomedical Optics

|October 5, 2016

Summary

This summary is machine-generated.

We developed a new algorithm and instrument for multiplexed fluorescence tomography, enabling simultaneous imaging of four fluorophores in tissue. This technique achieves high spatial resolution with zero cross-talk, advancing biomedical imaging capabilities.

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Area of Science:

Biomedical Optics
Fluorescence Imaging
Tomographic Reconstruction

Background:

Multiplexed fluorescence tomography is crucial for in vivo molecular imaging.
Current methods face challenges with spectral overlap and accurate reconstruction.
Simultaneous imaging of multiple fluorophores with high resolution remains a significant hurdle.

Purpose of the Study:

To develop and validate a novel algorithm and instrument for multiplexed fluorescence tomography.
To enable concurrent imaging of at least four fluorophores in the red and near-infrared spectrum.
To achieve high spatial resolution and eliminate image cross-talk in complex media.

Main Methods:

Developed a fluorescence tomography instrument acquiring spectral and temporal data.
Implemented an algorithm utilizing both spectral and temporal information for demixing.
Utilized extended fluorophore signature libraries to account for target positional variability.
Validated the system using tissue-mimicking phantoms with four spectrally overlapping fluorescent targets.

Main Results:

Successfully demixed and tomographically imaged four concurrent fluorophores.
Achieved zero image cross-talk between the fluorophores.
Demonstrated spatial resolution of 1 mm or better.
Validated performance in challenging tissue-mimicking phantom conditions.

Conclusions:

The developed algorithm and instrument enable robust multiplexed fluorescence tomography.
This technique overcomes spectral overlap limitations for multi-fluorophore imaging.
Offers a promising tool for high-resolution, cross-talk-free molecular imaging in biological tissues.