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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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Related Experiment Video

Updated: May 4, 2026

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Published on: April 9, 2014

Deep-tissue transcriptomics and subcellular imaging at high spatial resolution.

Valentina Gandin¹, Jun Kim¹, Liang-Zhong Yang¹

¹Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.

Science (New York, N.Y.)

|February 20, 2025

Summary

This summary is machine-generated.

New cycle hybridization chain reaction (cycleHCR) imaging overcomes limited fluorescence channels for detailed spatial analysis of RNA and proteins in biological specimens, enabling deep tissue research.

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Last Updated: May 4, 2026

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Author Spotlight: Investigating mRNA Spatial Distribution in Drosophila Muscle Tissue

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Area of Science:

Molecular Biology
Microscopy Techniques
Genomics

Background:

Traditional fluorescence microscopy is limited by the number of available color channels, restricting simultaneous spatial analysis of multiple targets.
Analyzing spatial organization of RNA and proteins in complex biological systems requires advanced imaging methods.

Purpose of the Study:

To introduce and validate cycle hybridization chain reaction (cycleHCR) as a novel method for highly multiplexed imaging of RNA and proteins.
To demonstrate the capability of cycleHCR for 3D spatial analysis in whole embryos and subcellular structures.

Main Methods:

Developed cycleHCR by integrating multicycle DNA barcoding with hybridization chain reaction (HCR).
Applied cycleHCR to whole-embryo transcriptomics imaging and combined it with expansion microscopy for subcellular resolution.
Utilized cycleHCR for multiplex RNA and protein imaging in mouse hippocampal slices.

Main Results:

Achieved precise 3D gene expression and cell fate mapping in whole embryos (~310 microm depth).
Revealed intricate subcellular structures (10 types) in mouse embryonic fibroblasts using cycleHCR and expansion microscopy.
Uncovered complex gene expression gradients and cell-type-specific nuclear variations in mouse hippocampal slices.

Conclusions:

cycleHCR provides a versatile and quantitative framework for elucidating spatial regulation in deep tissues.
This method overcomes limitations of traditional fluorescence microscopy for multiplexed imaging.
cycleHCR has significant potential for biological research and diagnostic applications.