GSD microscopy: Difference between revisions
Lexic 4712 (talk | contribs) added text from resolft article, added references, deleted stub warnings |
|||
| Line 1: | Line 1: | ||
[[Image:rittweger_gsd2.jpg|thumb|Comparison of resolution between standard confocal microscopy and GSD microscopy. Left: Confocal recording of vacancies in diamonds. Single spots cannot be separated. Right: GSD recording of the same location. Single vacancies are clearly visible. The size of the pointlike vacancies, corresponding to the microscope's resolution, is about 15nm.]] |
|||
{{Wikify|date=April 2010}}\r\n\r\n{{orphan|date=October 2008}} |
|||
Ground State Depletion Microscopy, or GSD Microscopy, is a technology similar to [[STED microscopy| Stimulated Emission Depletion Microscopy]]. It can sharpen the fluorescent spot significantly and overcome the diffraction barrier. It achieves this goal by quenching the excited states of the dye molecules at the outer part of the fluorescence spot to the triplet state or other long-lived states. Therefore the fluorescence of the outer part of the spot is inhibited, and the spatial resolution is improved. |
|||
'''G'''round '''S'''tate '''D'''epletion Microscopy, or GSD Microscopy, is an implementation of the [[RESOLFT]] concept. The method was proposed in 1995<ref>{{cite journal |author=Stefan W. Hell M. Kroug |title=Ground-state-depletion fluorescence microscopy: a concept for breaking the diffraction resolution limit |journal=Applied Physics B: Lasers and Optics |year=1995 |volume=60 |issue=5 |pages=495-497 |doi=10.1007/BF01081333 |url= http://www.springerlink.com/content/h1303753vl272v75/fulltext.pdf}}</ref> and experimentally demonstrated in 2007 <ref>{{cite journal |author=Stefan Bretschneider, Christian Eggeling, Stefan W. Hell |title=Breaking the diffraction barrier in fluorescence microscopy by optical shelving |journal=Physical Review Letters |year=2007 |volume=98 |issue=5 |pages=218103 |doi=10.1103/PhysRevLett.98.218103 |url=http://link.aps.org/doi/10.1103/PhysRevLett.98.218103}}</ref>. It is the second concept to overcome the diffraction barrier in far-field optical microscopy published by [[Stefan Hell]]. Using [[nitrogen-vacancy center]]s in diamonds a resolution of up to 7.8nm was achieved in 2009.<ref>{{cite journal |author=Eva Rittweger, Dominik Wildanger, Stefan W. Hell |title=Far-field fluorescence nanoscopy of diamond color centers by ground state depletion |journal= EPL, A Letters Journal Exploring the Frontiers of Physics |year=2009 |volume=86 |page=14001 |doi=10.1209/0295-5075/86/14001 |
|||
==External links== |
|||
|url=http://www.iop.org/EJ/article/0295-5075/86/1/14001/epl_86_1_14001.pdf}}</ref> |
|||
*[http://www.mpibpc.gwdg.de/abteilungen/200/STED.htm STED-Microscopy] |
|||
==Principle== |
|||
| ⚫ | |||
In GSD microscopy fluorescent markers are used. In one condition the marker can freely be excited from ground state an returns spontaneously via emission of a fluorescence photon. If light of appropriate wavelength is present additionally the dye can be excited to a long lifed dark state, i.e. a state where none fluorescence occurs. As long as the molecule is in the long lifed dark state (e.g. a triplet state), it cannot be excited from the ground state. Switching between these two states (bright and dark) by applying light all preconditions for the [[RESOLFT]] concept and subdiffraction imaging are fullfilled and images with very high resolution can be obtained. |
|||
{{tech-stub}} |
|||
The implementation in a microscope is very similar to [[STED microscopy| Stimulated Emission Depletion Microscopy]]. Using an appropriate ring-like focal spot for the light that brings the molecule into the dark state, the fluorescence can be quenched at the outer part of the focal spot. Therefore fluorescence only still takes place at the center of the microscope's focal spot and the spatial resolution is improved. |
|||
==References== |
|||
<references/> |
|||
==See also== |
|||
* [http://www.nanoscopy.de/ Department of NanoBiophotonics] at the Max-Planck-Institute for Biophysical Chemistry (Göttingen, Germany) |
|||
[[Category:Microscopy]] |
|||
| ⚫ | |||
Revision as of 11:48, 4 May 2010
Ground State Depletion Microscopy, or GSD Microscopy, is an implementation of the RESOLFT concept. The method was proposed in 1995[1] and experimentally demonstrated in 2007 [2]. It is the second concept to overcome the diffraction barrier in far-field optical microscopy published by Stefan Hell. Using nitrogen-vacancy centers in diamonds a resolution of up to 7.8nm was achieved in 2009.[3]
Principle
In GSD microscopy fluorescent markers are used. In one condition the marker can freely be excited from ground state an returns spontaneously via emission of a fluorescence photon. If light of appropriate wavelength is present additionally the dye can be excited to a long lifed dark state, i.e. a state where none fluorescence occurs. As long as the molecule is in the long lifed dark state (e.g. a triplet state), it cannot be excited from the ground state. Switching between these two states (bright and dark) by applying light all preconditions for the RESOLFT concept and subdiffraction imaging are fullfilled and images with very high resolution can be obtained.
The implementation in a microscope is very similar to Stimulated Emission Depletion Microscopy. Using an appropriate ring-like focal spot for the light that brings the molecule into the dark state, the fluorescence can be quenched at the outer part of the focal spot. Therefore fluorescence only still takes place at the center of the microscope's focal spot and the spatial resolution is improved.
References
- ^ Stefan W. Hell M. Kroug (1995). "Ground-state-depletion fluorescence microscopy: a concept for breaking the diffraction resolution limit" (PDF). Applied Physics B: Lasers and Optics. 60 (5): 495–497. doi:10.1007/BF01081333.
- ^ Stefan Bretschneider, Christian Eggeling, Stefan W. Hell (2007). "Breaking the diffraction barrier in fluorescence microscopy by optical shelving". Physical Review Letters. 98 (5): 218103. doi:10.1103/PhysRevLett.98.218103.
{{cite journal}}: CS1 maint: multiple names: authors list (link) - ^ Eva Rittweger, Dominik Wildanger, Stefan W. Hell (2009). "Far-field fluorescence nanoscopy of diamond color centers by ground state depletion" (PDF). EPL, A Letters Journal Exploring the Frontiers of Physics. 86: 14001. doi:10.1209/0295-5075/86/14001.
{{cite journal}}: CS1 maint: multiple names: authors list (link)
See also
- Department of NanoBiophotonics at the Max-Planck-Institute for Biophysical Chemistry (Göttingen, Germany)