The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function
Open accessAbstract
We demonstrate single-molecule fluorescence imaging beyond the optical diffraction limit in 3 dimensions with a wide-field microscope that exhibits a double-helix point spread function (DH-PSF). The DH-PSF design features high and uniform Fisher information and has 2 dominant lobes in the image plane whose angular orientation rotates with the axial ( z ) position of the emitter. Single fluorescent molecules in a thick polymer sample are localized in single 500-ms acquisitions with 10- to 20-nm precision over a large depth of field (2 μm) by finding the center of the 2 DH-PSF lobes. By using a photoactivatable fluorophore, repeated imaging of sparse subsets with a DH-PSF microscope provides superresolution imaging of high concentrations of molecules in all 3 dimensions. The combination of optical PSF design and digital postprocessing with photoactivatable fluorophores opens up avenues for improving 3D imaging resolution beyond the Rayleigh diffraction limit.
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Cites
Precise Nanometer Localization Analysis for Individual Fluorescent Probes
Thompson, R. E., Larson, D. R., & Webb, W. W. (2002) · Biophysical Journal · doi:10.1016/s0006-3495(02)75618-x · Cited by 2,320
Cited by
A guide to super-resolution fluorescence microscopy
Schermelleh, L., Heintzmann, R., & Leonhardt, H. (2010) · Journal of Cell Biology · doi:10.1083/jcb.201002018 · Cited by 1,255
Shim, S.-H., Xia, C., Zhong, G., Babcock, H. P., Vaughan, J. C., Huang, B., Wang, X., Xu, C., Bi, G.-Q., & Zhuang, X. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1201882109 · Cited by 514
Quantitative evaluation of software packages for single-molecule localization microscopy
Sage, D., Kirshner, H., Pengo, T., Stuurman, N., Min, J., Manley, S., & Unser, M. (2015) · Nature Methods · doi:10.1038/nmeth.3442 · Cited by 396
Single-molecule spectroscopy and imaging over the decades
Moerner, W. E., Shechtman, Y., & Wang, Q. (2015) · Faraday Discussions · doi:10.1039/c5fd00149h · Cited by 103
Self-citationMulticolour Single Molecule Imaging in Cells with Near Infra-Red Dyes
Tynan, C. J., Clarke, D. T., Coles, B. C., Rolfe, D. J., Martin-Fernandez, M. L., & Webb, S. E. D. (2012) · PLoS ONE · doi:10.1371/journal.pone.0036265 · Cited by 40
Metadata
- Authors
- Pavani, S. R. P., Thompson, M. A., Biteen, J. S., Lord, S. J., Liu, N., Twieg, R. J., Piestun, R., & Moerner, W. E.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2009-03-03
- Volume
- 106
- Issue
- 9
- Pages
- 2995-2999
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W4231350070
- Cited by
- 1,040
- References
- 1
- FWCI
- 30.56
- Type
- journal article
- Language
- en
- Open access
- green
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- Pavani, S. R. P., Thompson, M. A., Biteen, J. S., Lord, S. J., Liu, N., Twieg, R. J., Piestun, R., & Moerner, W. E. (2009). Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0900245106
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