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Exploiting the tunability of stimulated emission depletion microscopy for super-resolution imaging of nuclear structures.

Abstract：

:Imaging of nuclear structures within intact eukaryotic nuclei is imperative to understand the effect of chromatin folding on genome function. Recent developments of super-resolution fluorescence microscopy techniques combine high specificity, sensitivity, and less-invasive sample preparation procedures with the sub-diffraction spatial resolution required to image chromatin at the nanoscale. Here, we present a method to enhance the spatial resolution of a stimulated-emission depletion (STED) microscope based only on the modulation of the STED intensity during the acquisition of a STED image. This modulation induces spatially encoded variations of the fluorescence emission that can be visualized in the phasor plot and used to improve and quantify the effective spatial resolution of the STED image. We show that the method can be used to remove direct excitation by the STED beam and perform dual color imaging. We apply this method to the visualization of transcription and replication foci within intact nuclei of eukaryotic cells.

journal_name

Nat Commun

journal_title

Nature communications

authors

Sarmento MJ,Oneto M,Pelicci S,Pesce L,Scipioni L,Faretta M,Furia L,Dellino GI,Pelicci PG,Bianchini P,Diaspro A,Lanzanò L

doi

10.1038/s41467-018-05963-2

subject

Has Abstract

pub_date

2018-08-24 00:00:00

pages

3415

issue

1

issn

2041-1723

pii

10.1038/s41467-018-05963-2

journal_volume

9

pub_type

杂志文章

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