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Journal Article

Mapping molecules in scanning far-field fluorescence nanoscopy.

MPS-Authors
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Ta,  H.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Keller,  J.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Haltmeier,  M.
Research Group of Statistical Inverse-Problems in Biophysics, MPI for biophysical chemistry, Max Planck Society;

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Munk,  A.
Research Group of Statistical Inverse-Problems in Biophysics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

2181635.pdf
(Publisher version), 882KB

Supplementary Material (public)

2181635_Suppl_1.pdf
(Supplementary material), 3MB

2181635_Suppl_2.avi
(Supplementary material), 10MB

2181635_Suppl_3.zip
(Supplementary material), 51KB

Citation

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.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0028-374B-2
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.