Does Super-Resolution Fluorescence Microscopy Obsolete Previous Microscopic Approaches to Protein Co-localization?
| Autor: | Giulia Baldini, Laura J. MacDonald, Brian Storrie |
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| Rok vydání: | 2014 |
| Předmět: |
Microscopy
Confocal Materials science business.industry Super-resolution microscopy RESOLFT Resolution (electron density) STED microscopy Scanning confocal electron microscopy Proteins Article Protein Transport Optics Microscopy Fluorescence Light sheet fluorescence microscopy Microscopy Photoactivated localization microscopy business |
| Zdroj: | Membrane Trafficking ISBN: 9781493923083 |
| Popis: | Conventional microscopy techniques, namely, the confocal microscope or deconvolution processes, are resolution limited to approximately 200-250 nm by the diffraction properties of light as developed by Ernst Abbe in 1873. This diffraction limit is appreciably above the size of most multi-protein complexes, which are typically 20-50 nm in diameter. In the mid-2000s, biophysicists moved beyond the diffraction barrier by structuring the illumination pattern and then applying mathematical principles and algorithms to allow a resolution of approximately 100 nm, sufficient to address protein subcellular co-localization questions. This "breaking" of the diffraction barrier, affording resolution beyond 200 nm, is termed super-resolution microscopy. More recent approaches include single-molecule localization (such as photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM)) and point spread function engineering (such as stimulated emission depletion (STED) microscopy). In this review, we explain basic principles behind currently commercialized super-resolution setups and address advantages and considerations in applying these techniques to protein co-localization in biological systems. |
| Databáze: | OpenAIRE |
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