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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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Updated: Jun 14, 2025

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

Deep learning permits imaging of multiple structures with the same fluorophores.

Luhong Jin¹, Jingfang Liu², Heng Zhang²

¹School of Information Science and Technology, Hangzhou Normal University, Hangzhou, China; Department of Biomedical Engineering, MOE Key Laboratory of Biomedical Engineering, State Key Laboratory of Modern Optical Instrumentation, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, China.

Biophysical Journal

|September 5, 2024

Summary

This study introduces a novel double-structure network (DBSN) to overcome limitations in fluorescence microscopy. DBSN enables the extraction of six subcellular structures from just two fluorescent labels, improving live-cell imaging capabilities.

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Last Updated: Jun 14, 2025

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

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections

Published on: March 30, 2020

Area of Science:

Cell Biology
Microscopy Techniques
Bioimaging

Background:

Fluorescence microscopy is vital for observing cellular dynamics but limited by spectral overlap of dyes.
Current methods restrict the number of simultaneously imaged structures and are time-consuming due to sequential imaging.

Purpose of the Study:

To develop a novel computational method for enhanced multiplexed imaging in fluorescence microscopy.
To overcome the limitations of spectral overlap and sequential imaging in live-cell applications.

Main Methods:

A novel double-structure network (DBSN) was developed, integrating an intensity-balance model and a structure-separation model.
DBSN processes three raw images to extract six distinct subcellular structures using only two fluorescent labels.

Main Results:

The DBSN successfully extracts six distinct subcellular structures from limited fluorescent labels.
The intensity-balance model compensates for uneven fluorescence intensity across different structures.
The structure-separation model enables differentiation of structures labeled with the same fluorescent dye.

Conclusions:

The proposed DBSN significantly advances multiplexed imaging in fluorescence microscopy.
This method overcomes key bottlenecks in current live-cell imaging, offering broad potential in cell biology research.