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

0506542 - ÚFCH JH 2020 RIV GB eng J - Journal Article
Sarmento, Maria Joäo - Oneto, M. - Pelicci, S. - Pesce, L. - Scipioni, L. - Faretta, M. - Furia, L. - Dellino, G.I. - Pelicci, P.G. - Bianchini, P. - Diaspro, A. - Lanzano, L.
Exploiting the tunability of stimulated emission depletion microscopy for super-resolution imaging of nuclear structures.
Nature Communications. Roč. 9, AUG 2018 (2018), č. článku 3415. E-ISSN 2041-1723
Institutional support: RVO:61388955
Keywords : rna-polymerase-ii * illumination microscopy * chromosome structure * chromatin-structure * sted microscopy * human-cells * dna
OECD category: Physical chemistry
Impact factor: 11.878, year: 2018
Method of publishing: Open access

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.
Permanent Link: http://hdl.handle.net/11104/0297773