scholarly journals Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy

10.3791/51100 ◽  
2014 ◽  
Author(s):  
Emily M. Mace ◽  
Jordan S. Orange
Keyword(s):  
Immunological Synapse ◽  
Stimulated Emission ◽  
Dual Color ◽  
Stimulated Emission Depletion

Integrated dual-color stimulated emission depletion (STED) microscopy and fluorescence emission difference (FED) microscopy

2018 ◽  
Vol 423 ◽  
pp. 167-174 ◽  
Author(s):  
Wensheng Wang ◽  
Guangyuan Zhao ◽  
Cuifang Kuang ◽  
Liang Xu ◽  
Shaocong Liu ◽  
...  
Keyword(s):  
Fluorescence Emission ◽  
Stimulated Emission ◽  
Sted Microscopy ◽  
Dual Color ◽  
Stimulated Emission Depletion

scholarly journals Nanoscale distribution of nuclear sites analyzed by superresolution STED-ICCS

2019 ◽  
Author(s):  
M. Oneto ◽  
L. Scipioni ◽  
M.J. Sarmento ◽  
I. Cainero ◽  
S. Pelicci ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Cross Correlation ◽  
Correlation Spectroscopy ◽  
Stimulated Emission ◽  
Nuclear Antigen ◽  
Dynamic Interactions ◽  
Functional Sites ◽  
Dual Color ◽  
Replication Foci ◽  
Stimulated Emission Depletion

AbstractDeciphering the spatiotemporal coordination between nuclear functions is important to understand its role in the maintenance of human genome. In this context, superresolution microscopy has gained considerable interest as it can be used to probe the spatial organization of functional sites in intact single cell nuclei in the 20-250 nm range. Among the methods that quantify colocalization from multicolor images, image cross-correlation spectroscopy (ICCS) offers several advantages, namely it does not require a pre-segmentation of the image into objects and can be used to detect dynamic interactions. However, the combination of ICCS with super-resolution microscopy has not been explored yet.Here we combine dual color stimulated emission depletion (STED) nanoscopy with ICCS (STED-ICCS) to quantify the nanoscale distribution of functional nuclear sites. We show that STED-ICCS provides not only a value of colocalized fraction but also the characteristic distances associated to correlated nuclear sites. As a validation, we quantify the nanoscale spatial distribution of three different pairs of functional nuclear sites in MCF10A cells. As expected, transcription foci and a transcriptionally repressive histone marker (H3K9me3) are not correlated. Conversely, nascent DNA replication foci and the Proliferating cell nuclear antigen (PCNA) protein have a high level of proximity and are correlated at a nanometer distance which is close to the limit of our experimental approach. Finally, transcription foci are found at a distance of 130 nm from replication foci, indicating a spatial segregation at the nanoscale. Overall, our data demonstrate that STED-ICCS can be a powerful tool for the analysis of nanoscale distribution of functional sites in the nucleus.Statement of significanceSeveral methods are available to quantify the proximity of two labeled molecules from dual color images. Among them, image cross-correlation spectroscopy (ICCS) is attractive as it does not require a pre-segmentation of the image into objects and can be used to detect dynamic interactions. Here, we combine for the first time ICCS with superresolution stimulated emission depletion (STED) microscopy (STED-ICCS) to quantify the spatial distribution of functional sites in the nucleus. Our results show that STED-ICCS, in addition to quantifying the colocalized fraction, detects characteristic nanometer distances associated to correlated nuclear sites. This work shows that STED-ICCS can be a powerful tool to quantify the nanoscale distribution of functional sites in the nucleus.

scholarly journals Stimulated Emission Depletion Microscopy with Color Centers in Hexagonal Boron Nitride

ACS Photonics ◽  
2021 ◽  
Author(s):  
Prince Khatri ◽  
Ralph Nicholas Edward Malein ◽  
Andrew J. Ramsay ◽  
Isaac J. Luxmoore
Keyword(s):  
Boron Nitride ◽  
Hexagonal Boron Nitride ◽  
Stimulated Emission ◽  
Color Centers ◽  
Stimulated Emission Depletion

scholarly journals Stimulated Emission Depletion Microscopy Resolves Individual Nitrogen Vacancy Centers in Diamond Nanocrystals

ACS Nano ◽  
2013 ◽  
Vol 7 (12) ◽  
pp. 10912-10919 ◽  
Author(s):  
Silvia Arroyo-Camejo ◽  
Marie-Pierre Adam ◽  
Mondher Besbes ◽  
Jean-Paul Hugonin ◽  
Vincent Jacques ◽  
...  
Keyword(s):  
Stimulated Emission ◽  
Nitrogen Vacancy ◽  
Nitrogen Vacancy Centers ◽  
Stimulated Emission Depletion
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