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  Mapping molecules in scanning far-field fluorescence nanoscopy.

Ta, H., Keller, J., Haltmeier, M., Saka, S. K., Schmied, J., Opazo, F., et al. (2015). Mapping molecules in scanning far-field fluorescence nanoscopy. Nature Communications, 6: 7977. doi:10.1038/ncomms8977.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-374B-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-285F-9
Genre: Journal Article

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 Creators:
Ta, H.1, Author              
Keller, J.1, Author              
Haltmeier, M.2, Author              
Saka, S. K., Author
Schmied, J., Author
Opazo, F., Author
Tinnefeld, P., Author
Munk, A.2, Author              
Hell, S. W.1, Author              
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              
2Research Group of Statistical Inverse-Problems in Biophysics, MPI for biophysical chemistry, Max Planck Society, ou_1113580              

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 Abstract: In fluorescence microscopy, the distribution of the emitting molecule number in space is usually obtained by dividing the measured fluorescence by that of a single emitter. However, the brightness of individual emitters may vary strongly in the sample or be inaccessible. Moreover, with increasing (super-) resolution, fewer molecules are found per pixel, making this approach unreliable. Here we map the distribution of molecules by exploiting the fact that a single molecule emits only a single photon at a time. Thus, by analysing the simultaneous arrival of multiple photons during confocal imaging, we can establish the number and local brightness of typically up to 20 molecules per confocal (diffraction sized) recording volume. Subsequent recording by stimulated emission depletion microscopy provides the distribution of the number of molecules with subdiffraction resolution. The method is applied to mapping the three-dimensional nanoscale organization of internalized transferrin receptors on human HEK293 cells.

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Language(s): eng - English
 Dates: 2015-08-13
 Publication Status: Published online
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1038/ncomms8977
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Title: Nature Communications
Source Genre: Journal
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Pages: 7 Volume / Issue: 6 Sequence Number: 7977 Start / End Page: - Identifier: -