scholarly journals Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

10.3791/54774 ◽  
2016 ◽  
Author(s):  
Yilin Wang ◽  
Pakorn Kanchanawong
Keyword(s):  
Actin Filaments ◽  
Three Dimensional ◽  
Super Resolution ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy ◽  
Super Resolution Microscopy

scholarly journals Three Dimensional Super Resolution Fluorescence Imaging of Single Bacterial Cells by Stereo Photoactivated Localization Microscopy

2009 ◽  
Vol 15 (S2) ◽  
pp. 564-565 ◽  
Author(s):  
J Tang ◽  
A Vaziri ◽  
J Akerboom ◽  
L Looger ◽  
C Shank
Keyword(s):  
Fluorescence Imaging ◽  
Three Dimensional ◽  
Super Resolution ◽  
Bacterial Cells ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

Biological Insight from Super-Resolution Microscopy: What We Can Learn from Localization-Based Images

2018 ◽  
Vol 87 (1) ◽  
pp. 965-989 ◽  
Author(s):  
David Baddeley ◽  
Joerg Bewersdorf
Keyword(s):  
Imaging Modality ◽  
Super Resolution ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy ◽  
Stochastic Optical Reconstruction Microscopy ◽  
Biological Insight ◽  
Biological Discovery ◽  
Optical Reconstruction ◽  
Super Resolution Microscopy ◽  
Fluorescent Molecules

Super-resolution optical imaging based on the switching and localization of individual fluorescent molecules [photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), etc.] has evolved remarkably over the last decade. Originally driven by pushing technological limits, it has become a tool of biological discovery. The initial demand for impressive pictures showing well-studied biological structures has been replaced by a need for quantitative, reliable data providing dependable evidence for specific unresolved biological hypotheses. In this review, we highlight applications that showcase this development, identify the features that led to their success, and discuss remaining challenges and difficulties. In this context, we consider the complex topic of defining resolution for this imaging modality and address some of the more common analytical methods used with this data.

scholarly journals Enlightening G-protein-coupled receptors on the plasma membrane using super-resolution photoactivated localization microscopy

2013 ◽  
Vol 41 (1) ◽  
pp. 191-196 ◽  
Author(s):  
Marco Scarselli ◽  
Paolo Annibale ◽  
Claudio Gerace ◽  
Aleksandra Radenovic
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
Super Resolution ◽  
Molecular Size ◽  
G Protein Coupled Receptors ◽  
Temporal Organization ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy ◽  
Super Resolution Microscopy ◽  
G Protein Coupled

The possibility to visualize and image the arrangement of proteins within the cell at the molecular level has always been an attraction for scientists in biological research. In particular, for signalling molecules such as GPCRs (G-protein-coupled receptors), the existence of protein aggregates such as oligomers or clusters has been the topic of extensive debate. One of the reasons for this lively argument is that the molecular size is below the diffraction-limited resolution of the conventional microscopy, precluding the direct visualization of protein super-structures. On the other hand, new super-resolution microscopy techniques, such as the PALM (photoactivated localization microscopy), allow the limit of the resolution power of conventional optics to be broken and the localization of single molecules to be determined with a precision of 10–20 nm, close to their molecular size. The application of super-resolution microscopy to study the spatial and temporal organization of GPCRs has brought new insights into receptor arrangement on the plasma membrane. Furthermore, the use of this powerful microscopy technique as a quantitative tool opens up the possibility for investigating and quantifying the number of molecules in biological assemblies and determining the protein stoichiometry in signalling complexes.

scholarly journals Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes

2021 ◽  
Vol 22 (4) ◽  
pp. 1903
Author(s):  
Ivona Kubalová ◽  
Alžběta Němečková ◽  
Klaus Weisshart ◽  
Eva Hřibová ◽  
Veit Schubert
Keyword(s):  
Single Molecule ◽  
Imaging Techniques ◽  
Chromatin Condensation ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Wide Field ◽  
Structured Illumination Microscopy ◽  
Metaphase Chromosomes ◽  
Localization Microscopy ◽  
Super Resolution Microscopy

The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200–250 nm laterally, ~500–700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4′,6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution.

Cubic spline-based depth-dependent localization of mitochondria–endoplasmic reticulum contacts by three-dimensional light-sheet super-resolution microscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Yucheng Sun ◽  
Seungah Lee ◽  
Seong Ho Kang
Keyword(s):  
Endoplasmic Reticulum ◽  
Calcium Homeostasis ◽  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet ◽  
Cubic Spline ◽  
Contact Distance ◽  
Super Resolution Microscopy

The contact distance between mitochondria (Mito) and endoplasmic reticulum (ER) has received considerable attention owing to their crucial function in maintaining lipid and calcium homeostasis. Herein, cubic spline algorithm-based depth-dependent...

Super-Resolution Microscopy: SIM, STED and Localization Microscopy

2015 ◽  
pp. 47-60 ◽  
Author(s):  
James Dodgson ◽  
Anatole Chessel ◽  
Susan Cox ◽  
Rafael E. Carazo Salas
Keyword(s):  
Super Resolution ◽  
Localization Microscopy ◽  
Super Resolution Microscopy

scholarly journals Stereo Photoactivated Localization Microscopy for Super-Resolution 3D Bioimaging

2009 ◽  
Vol 96 (3) ◽  
pp. 16a
Author(s):  
Jianyong Tang ◽  
Alipasha Vaziri ◽  
Charles V. Shank
Keyword(s):  
Super Resolution ◽  
Localization Microscopy ◽  
Photoactivated Localization Microscopy

scholarly journals Small-Molecule Labeling of Live Cell Surfaces for Three-Dimensional Super-Resolution Microscopy

2014 ◽  
Vol 136 (40) ◽  
pp. 14003-14006 ◽  
Author(s):  
Marissa K. Lee ◽  
Prabin Rai ◽  
Jarrod Williams ◽  
Robert J. Twieg ◽  
W. E. Moerner
Keyword(s):  
Small Molecule ◽  
Three Dimensional ◽  
Super Resolution ◽  
Live Cell ◽  
Cell Surfaces ◽  
Super Resolution Microscopy
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