In Situ Single-Molecule Imaging with Attoliter Detection Using Objective Total Internal Reflection Confocal Microscopy†

Biochemistry ◽  
2006 ◽  
Vol 45 (13) ◽  
pp. 4058-4068 ◽  
Author(s):  
Thomas P. Burghardt ◽  
Katalin Ajtai ◽  
Julian Borejdo
Keyword(s):  
Confocal Microscopy ◽  
Single Molecule ◽  
Total Internal Reflection ◽  
Internal Reflection ◽  
Single Molecule Imaging

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 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.

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