scholarly journals Correlative super-resolution fluorescence and electron microscopy using conventional fluorescent proteins in vacuo

2017 ◽  
Vol 199 (2) ◽  
pp. 120-131 ◽  
Author(s):  
Christopher J. Peddie ◽  
Marie-Charlotte Domart ◽  
Xenia Snetkov ◽  
Peter O'Toole ◽  
Banafshe Larijani ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Fluorescent Proteins ◽  
Super Resolution ◽  
Fluorescence And Electron Microscopy

scholarly journals Erratum: Corrigendum: Correlative in-resin super-resolution and electron microscopy using standard fluorescent proteins

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Errin Johnson ◽  
Elena Seiradake ◽  
E. Yvonne Jones ◽  
Ilan Davis ◽  
Kay Grünewald ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Fluorescent Proteins ◽  
Super Resolution

scholarly journals A Cyclometalated Iridium (III) Complex as a Microtubule Probe for Correlative Super‐Resolution Fluorescence and Electron Microscopy

2020 ◽  
Vol 32 (39) ◽  
pp. 2003901
Author(s):  
Xiaohe Tian ◽  
Cesare De Pace ◽  
Lorena Ruiz‐Perez ◽  
Bo Chen ◽  
Rina Su ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Super Resolution ◽  
Fluorescence And Electron Microscopy

scholarly journals Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution

2014 ◽  
Vol 127 (20) ◽  
pp. 4351-4355 ◽  
Author(s):  
Anna Löschberger ◽  
Christian Franke ◽  
Georg Krohne ◽  
Sebastian van de Linde ◽  
Markus Sauer
Keyword(s):  
Electron Microscopy ◽  
Nuclear Pore Complex ◽  
Super Resolution ◽  
Nuclear Pore ◽  
Fluorescence And Electron Microscopy

scholarly journals Correlative in-resin super-resolution and electron microscopy using standard fluorescent proteins

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Errin Johnson ◽  
Elena Seiradake ◽  
E. Yvonne Jones ◽  
Ilan Davis ◽  
Kay Grünewald ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Single Molecule ◽  
Fluorescent Proteins ◽  
Super Resolution ◽  
Freeze Substitution ◽  
Microscopy Imaging ◽  
Localization Accuracy ◽  
Culture Cells ◽  
Structural Preservation ◽  
Key Aspects

Abstract We introduce a method for correlative in-resin super-resolution fluorescence and electron microscopy (EM) of biological structures in mammalian culture cells. Cryo-fixed resin embedded samples offer superior structural preservation, performing in-resin super-resolution, however, remains a challenge. We identified key aspects of the sample preparation procedure of high pressure freezing, freeze substitution and resin embedding that are critical for preserving fluorescence and photo-switching of standard fluorescent proteins, such as mGFP, mVenus and mRuby2. This enabled us to combine single molecule localization microscopy with transmission electron microscopy imaging of standard fluorescent proteins in cryo-fixed resin embedded cells. We achieved a structural resolution of 40–50 nm (~17 nm average single molecule localization accuracy) in the fluorescence images without the use of chemical fixation or special fluorophores. Using this approach enabled the correlation of fluorescently labeled structures to the ultrastructure in the same cell at the nanometer level and superior structural preservation.

scholarly journals VIPER is a genetically encoded peptide tag for fluorescence and electron microscopy

2018 ◽  
Vol 115 (51) ◽  
pp. 12961-12966 ◽  
Author(s):  
Julia K. Doh ◽  
Jonathan D. White ◽  
Hannah K. Zane ◽  
Young Hwan Chang ◽  
Claudia S. López ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Biology ◽  
Iron Uptake ◽  
Fluorescent Proteins ◽  
Coiled Coil ◽  
Protein Targets ◽  
Cellular Environment ◽  
Fluorescence And Electron Microscopy ◽  
Specific Proteins ◽  
Peptide Tag

Many discoveries in cell biology rely on making specific proteins visible within their native cellular environment. There are various genetically encoded tags, such as fluorescent proteins, developed for fluorescence microscopy (FM). However, there are almost no genetically encoded tags that enable cellular proteins to be observed by both FM and electron microscopy (EM). Herein, we describe a technology for labeling proteins with diverse chemical reporters, including bright organic fluorophores for FM and electron-dense nanoparticles for EM. Our technology uses versatile interacting peptide (VIP) tags, a class of genetically encoded tag. We present VIPER, which consists of a coiled-coil heterodimer formed between the genetic tag, CoilE, and a probe-labeled peptide, CoilR. Using confocal FM, we demonstrate that VIPER can be used to highlight subcellular structures or to image receptor-mediated iron uptake. Additionally, we used VIPER to image the iron uptake machinery by correlative light and EM (CLEM). VIPER compared favorably with immunolabeling for imaging proteins by CLEM, and is an enabling technology for protein targets that cannot be immunolabeled. VIPER is a versatile peptide tag that can be used to label and track proteins with diverse chemical reporters observable by both FM and EM instrumentation.

scholarly journals Correlative cryo super-resolution light and electron microscopy on mammalian cells using fluorescent proteins

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Maarten W. Tuijtel ◽  
Abraham J. Koster ◽  
Stefan Jakobs ◽  
Frank G. A. Faas ◽  
Thomas H. Sharp
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Fluorescent Proteins ◽  
Light And Electron Microscopy ◽  
Super Resolution
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