scholarly journals Introduction: Modern Imaging in Biology and Medicine: Papers from the Seventh Omaha Imaging Symposium, April 2011

2012 ◽  
Vol 18 (4) ◽  
pp. 728-729
Author(s):  
Richard Hallworth ◽  
Michael G. Nichols
Keyword(s):  
Confocal Microscopy ◽  
Single Molecule ◽  
Total Internal Reflection ◽  
Imaging Techniques ◽  
Special Section ◽  
Biological Imaging ◽  
Internal Reflection ◽  
Biological Research ◽  
Student Life ◽  
Multiphoton Excitation

The ability to see, or visualize, a phenomenon is an essential tool of modern biological research. Our ability to create static and dynamic images has grown exponentially in the 25 or so years since confocal microscopy became readily available. The ingenious and energetic application of insights from optical physics to biological imaging in recent years has brought us far-reaching extensions of simple imaging, including nonlinear (or multiphoton) excitation, total internal reflection imaging, and even single molecule counting techniques. The annual Omaha Imaging Symposium has since 2003 brought together experts in advanced biological imaging techniques for a one-day exposition of how these techniques help move biological science forward. The seventh iteration of the series was held Friday, April 8, 2011, at the Harper Student Life Center of Creighton University, in Omaha, Nebraska. This special section of Microscopy and Micronanalysis consists of papers from speakers at the symposium.

scholarly journals Introduction: New Horizons in Research Imaging. Papers from the Eighth Omaha Imaging Symposium, April 2012

2013 ◽  
Vol 19 (4) ◽  
pp. 777-777
Author(s):  
Richard Hallworth
Keyword(s):  
Imaging Techniques ◽  
Special Section ◽  
Biological Imaging ◽  
Biological Research ◽  
Student Life ◽  
New Horizons

Since 2003, the annual Omaha Imaging Symposium has brought together experts in advanced biological imaging techniques for a one-day exposition of how advanced research imaging techniques foster progress in biological research. The eighth symposium in the series was held Friday, April 20, 2012, at the Harper Student Life Center of Creighton University, Omaha, Nebraska. This special section of Microscopy and Microanalysis consists of articles contributed by speakers at the symposium.

scholarly journals Three-dimensional total-internal reflection fluorescence nanoscopy with nanometric axial resolution by photometric localization of single molecules

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Alan M. Szalai ◽  
Bruno Siarry ◽  
Jerónimo Lukin ◽  
David J. Williamson ◽  
Nicolás Unsain ◽  
...  
Keyword(s):  
Single Molecule ◽  
Total Internal Reflection ◽  
Single Molecules ◽  
Fluorescence Microscope ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Axial Position ◽  
Localization Microscopy ◽  
Localization Precision ◽  
Single Molecule Localization Microscopy

AbstractSingle-molecule localization microscopy enables far-field imaging with lateral resolution in the range of 10 to 20 nanometres, exploiting the fact that the centre position of a single-molecule’s image can be determined with much higher accuracy than the size of that image itself. However, attaining the same level of resolution in the axial (third) dimension remains challenging. Here, we present Supercritical Illumination Microscopy Photometric z-Localization with Enhanced Resolution (SIMPLER), a photometric method to decode the axial position of single molecules in a total internal reflection fluorescence microscope. SIMPLER requires no hardware modification whatsoever to a conventional total internal reflection fluorescence microscope and complements any 2D single-molecule localization microscopy method to deliver 3D images with nearly isotropic nanometric resolution. Performance examples include SIMPLER-direct stochastic optical reconstruction microscopy images of the nuclear pore complex with sub-20 nm axial localization precision and visualization of microtubule cross-sections through SIMPLER-DNA points accumulation for imaging in nanoscale topography with sub-10 nm axial localization precision.

Super-Resolution Imaging in Fluid Mechanics Using New Illumination Approaches

2020 ◽  
Vol 52 (1) ◽  
pp. 369-393
Author(s):  
Minami Yoda
Keyword(s):  
Fluid Mechanics ◽  
Total Internal Reflection ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Internal Reflection ◽  
Interfacial Flows ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Entire Volume ◽  
Resolution Imaging

Quantifying submillimeter flows using optical diagnostic techniques is often limited by a lack of spatial resolution and optical access. This review discusses two super-resolution imaging techniques, structured illumination microscopy and total internal reflection fluorescence or microscopy, which can visualize bulk and interfacial flows, respectively, at spatial resolutions below the classic diffraction limits. First, we discuss the theory and applications of structured illumination for optical sectioning, i.e., imaging a thin slice of a flow illuminated over its entire volume. Structured illumination can be used to visualize the interior of multiphase flows such as sprays by greatly reducing secondary scattering. Second, the theory underlying evanescent waves is introduced, followed by a review of how total internal reflection microscopy has been used to visualize interfacial flows over the last 15 years. Both techniques, which are starting to be used in fluid mechanics, could significantly improve quantitative imaging of microscale and macroscale flows.

scholarly journals Total Internal Reflection Fluorescence Microscopy (TIRFM) – novel techniques and applications

2020 ◽  
Vol 8 (11) ◽  
Author(s):  
Verena Richter ◽  
Michael Wagner ◽  
Herbert Schneckenburger
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Total Internal Reflection ◽  
Plasma Membranes ◽  
Focal Adhesions ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Thin Layers ◽  
Structured Illumination ◽  
Structured Illumination Microscopy

Total Internal Reflection Fluorescence Microscopy (TIRFM) has been established almost 40 years ago for studies of plasma membranes or membrane proximal sites of living cells. The method is based on light incidence at an angle above the critical angle of total internal reflection and generation of an evanescent electromagnetic field penetrating about 100 nm into a sample and permitting selective excitation of membrane proximal fluorophores. Two techniques are presented here: prism-type TIRFM and objective-type TIRFM with high aperture microscope objective lenses. Furthermore, numerous applications are summarized, e.g. measurement of focal adhesions, cell-substrate topology, endocytosis or exocytosis of vesicles as well as single molecule detection within thin layers. Finally, highly innovative combinations of TIRFM with Förster Resonance Energy Transfer (FRET) measurements as well as with Structured Illumination Microscopy (SIM) and fluorescence reader technologies are presented.

scholarly journals Single-Molecule Total Internal Reflection Fluorescence Microscopy

2016 ◽  
Vol 2016 (5) ◽  
pp. pdb.top077800 ◽  
Author(s):  
Emily M. Kudalkar ◽  
Trisha N. Davis ◽  
Charles L. Asbury
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence

scholarly journals Severing enzymes amplify microtubule arrays through lattice GTP-tubulin incorporation

Science ◽  
2018 ◽  
Vol 361 (6404) ◽  
pp. eaau1504 ◽  
Author(s):  
Annapurna Vemu ◽  
Ewa Szczesna ◽  
Elena A. Zehr ◽  
Jeffrey O. Spector ◽  
Nikolaus Grigorieff ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
High Density ◽  
Guanosine Triphosphate ◽  
Microtubule Severing

Spastin and katanin sever and destabilize microtubules. Paradoxically, despite their destructive activity they increase microtubule mass in vivo. We combined single-molecule total internal reflection fluorescence microscopy and electron microscopy to show that the elemental step in microtubule severing is the generation of nanoscale damage throughout the microtubule by active extraction of tubulin heterodimers. These damage sites are repaired spontaneously by guanosine triphosphate (GTP)–tubulin incorporation, which rejuvenates and stabilizes the microtubule shaft. Consequently, spastin and katanin increase microtubule rescue rates. Furthermore, newly severed ends emerge with a high density of GTP-tubulin that protects them against depolymerization. The stabilization of the newly severed plus ends and the higher rescue frequency synergize to amplify microtubule number and mass. Thus, severing enzymes regulate microtubule architecture and dynamics by promoting GTP-tubulin incorporation within the microtubule shaft.

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