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Fluorescence Imaging with One-nanometer Accuracy (FIONA)
11:56

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

Fluorescence Imaging with One-Nanometer Accuracy (FIONA).

Paul R Selvin1, Tyler Lougheed, Melinda Tonks Hoffman

  • 1Biophysics Center and Physics Department, University of Illinois, Urbana, IL 61801, USA.

CSH Protocols
|March 2, 2011
PubMed
Summary

Fluorescence imaging with one-nanometer accuracy (FIONA) offers precise single-molecule localization. Variants like SHRIMP, SHREC, DOPI, and bFIONA extend its capabilities for static, dynamic, 3D, and non-fluorescent imaging.

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Fluorescence Imaging with One-nanometer Accuracy (FIONA)
11:56

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

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08:32

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Published on: October 28, 2018

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Nanotechnology

Background:

  • Single-molecule localization techniques are crucial for understanding molecular dynamics.
  • Fluorescence imaging with one-nanometer accuracy (FIONA) provides high-precision localization of single dye molecules.
  • Existing FIONA variants have limitations regarding system dynamics or dye requirements.

Purpose of the Study:

  • To review and compare various advanced FIONA techniques.
  • To highlight the capabilities of different FIONA variants for specific applications.
  • To discuss the potential of non-fluorescent imaging methods.

Main Methods:

  • Total internal reflection fluorescence microscopy
  • Deoxygenation agents and high quantum yield detectors for FIONA
  • Single-molecule high-resolution imaging with photobleaching (SHRIMP) for static systems
  • Single-molecule high-resolution colocalization (SHREC) for dynamic systems
  • Defocused orientation and positional imaging (DOPI) for 3D orientation and localization
  • Bright-field imaging with one-nanometer accuracy (bFIONA) for non-fluorescent imaging

Main Results:

  • FIONA achieves ~1-nm localization accuracy.
  • SHRIMP offers high resolution for static systems using a single dye type.
  • SHREC extends high-resolution imaging to dynamic systems.
  • DOPI determines 3D orientation and achieves few-nanometer localization.
  • bFIONA provides FIONA-like localization without fluorescence.

Conclusions:

  • FIONA and its variants offer powerful tools for nanoscale imaging.
  • Specific FIONA techniques are suited for different biological and physical systems.
  • bFIONA presents an alternative for situations where fluorescence is not feasible.