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Super-resolution Fluorescence Microscopy

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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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Recurrent Herpetic Stromal Keratitis in Mice, a Model for Studying Human HSK

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Published on: December 18, 2012

Visualizing the replicating HSV-1 virus using STED super-resolution microscopy.

Zhuoran Li^1,2, Ce Fang³, Yuanyuan Su³

¹Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, NO. 32 Jiaochang Donglu, Kunming, Yunnan, 650223, People's Republic of China.

Virology Journal

|April 11, 2016

Summary

This summary is machine-generated.

Super-resolution microscopy reveals herpes simplex virus 1 (HSV-1) genome expansion during replication. Viral replication and transcription are spatially separated within replication compartments, offering new insights into HSV-1 infection dynamics.

Keywords:

FISH HSV-1 replication ICP8 IF RNA Pol II STED

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Published on: April 9, 2017

Area of Science:

Virology
Microscopy
Molecular Biology

Background:

Herpes simplex virus 1 (HSV-1) lytic infection involves viral genome replication, but details remain unclear due to microscopy limitations.
Stimulated emission depletion (STED) microscopy offers super-resolution imaging for enhanced visualization of viral processes.

Purpose of the Study:

To investigate the dynamics of HSV-1 genome replication using advanced microscopy techniques.
To elucidate the spatial organization of viral replication and transcription within infected cells.

Main Methods:

Utilized stimulated emission depletion (STED) microscopy combined with fluorescence in situ hybridization (FISH) and immunofluorescence (IF).
Employed dual-color probes to label specific regions of the HSV-1 genome.
Visualized viral proteins, including ICP8, and host factors like RNA polymerase II.

Main Results:

HSV-1 genome undergoes spatial expansion from a compact to a relaxed state during replication.
Viral single-strand binding protein ICP8 is closely associated with replicating HSV-1 DNA.
Replication and transcription sites are spatially segregated within viral replication compartments, with ICP8-marked replication sites separated from RNA polymerase II.

Conclusions:

Super-resolution microscopy enables observation of replicating HSV-1 genomes.
The HSV-1 genome spatially expands during the replication process.
Viral replication and transcription are compartmentalized within distinct sub-structures.