The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.
Confined activation and subdiffractive localization enables whole-cell PALM with genetically expressed probes
Open accessAbstract
We demonstrate three-dimensional (3D) super-resolution microscopy in whole fixed cells using photoactivated localization microscopy (PALM). The use of the bright, genetically expressed fluorescent marker photoactivatable monomeric (m)Cherry (PA-mCherry1) in combination with near diffraction-limited confinement of photoactivation using two-photon illumination and 3D localization methods allowed us to investigate a variety of cellular structures at <50 nm lateral and <100 nm axial resolution. Compared to existing methods, we have substantially reduced excitation and bleaching of unlocalized markers, which allows us to use 3D PALM imaging with high localization density in thick structures. Our 3D localization algorithms, which are based on cross-correlation, do not rely on idealized noise models or specific optical configurations. This allows instrument design to be flexible. By generating appropriate fusion constructs and expressing them in Cos7 cells, we could image invaginations of the nuclear membrane, vimentin fibrils, the mitochondrial network and the endoplasmic reticulum at depths of greater than 8 μm.
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Cites
Matplotlib: a 2D graphics environment
Hunter, J. D. (2007) · Computing in Science & Engineering · doi:10.1109/mcse.2007.55 · Cited by 39,249
Python for scientific computing
Oliphant, T. E. (2007) · Computing in Science & Engineering · doi:10.1109/mcse.2007.58 · Cited by 3,303 · cited 1×
“Our algorithm works with different imaging configurations (for example, with or without a cylindrical lens) and is implemented in a free, platform-independent languag”
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Cited by
Precisely and accurately localizing single emitters in fluorescence microscopy
Deschout, H., Zanacchi, F. C., Mlodzianoski, M., Diaspro, A., Bewersdorf, J., Hess, S. T., & Braeckmans, K. (2014) · Nature Methods · doi:10.1038/nmeth.2843 · Cited by 498
York, A. G., Parekh, S. H., Nogare, D. D., Fischer, R. S., Temprine, K., Mione, M., Chitnis, A. B., Combs, C. A., & Shroff, H. (2012) · Nature Methods · doi:10.1038/nmeth.2025 · Cited by 472
Self-citationMartínez-Mármol, R., Giordano-Santini, R., Kaulich, E., Cho, A.-N., Przybyla, M., Riyadh, M. A., Robinson, E., Chew, K. Y., Amor, R., Meunier, F. A., et al. (2023) · Science Advances · doi:10.1126/sciadv.adg2248 · Cited by 94
Metadata
- Authors
- York, A. G., Ghitani, A., Vaziri, A., Davidson, M. W., & Shroff, H.
- Venue
- Nature Methods
- Published
- 2011-02-13
- Volume
- 8
- Issue
- 4
- Pages
- 327-333
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2075924762
- Cited by
- 172
- References
- 2
- FWCI
- 16.66
- Type
- journal article
- Language
- en
- Open access
- green
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- York, A. G., Ghitani, A., Vaziri, A., Davidson, M. W., & Shroff, H. (2011). Confined activation and subdiffractive localization enables whole-cell PALM with genetically expressed probes. Nature Methods. https://doi.org/10.1038/nmeth.1571
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