Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy
The morphological features of α-synuclein (AS) amyloid aggregation in vitro and in cells were elucidated at the nanoscale by far-field subdiffraction fluorescence localization microscopy. Labeling AS with rhodamine spiroamide probes allowed us to image AS fibrillar structures by fluorescence stochas...
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todo:paper_00063495_v102_n7_p1598_Roberti2023-10-03T14:05:02Z Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy Roberti, M.J. Fölling, J. Celej, M.S. Bossi, M. Jovin, T.M. Jares-Erijman, E.A. alpha synuclein nanomaterial rhodamine article chemistry color fluorescence microscopy HeLa cell human intracellular space metabolism methodology molecular imaging protein multimerization protein secondary structure alpha-Synuclein Color HeLa Cells Humans Intracellular Space Microscopy, Fluorescence Molecular Imaging Nanostructures Protein Multimerization Protein Structure, Secondary Rhodamines The morphological features of α-synuclein (AS) amyloid aggregation in vitro and in cells were elucidated at the nanoscale by far-field subdiffraction fluorescence localization microscopy. Labeling AS with rhodamine spiroamide probes allowed us to image AS fibrillar structures by fluorescence stochastic nanoscopy with an enhanced resolution at least 10-fold higher than that achieved with conventional, diffraction-limited techniques. The implementation of dual-color detection, combined with atomic force microscopy, revealed the propagation of individual fibrils in vitro. In cells, labeled protein appeared as amyloid aggregates of spheroidal morphology and subdiffraction sizes compatible with in vitro supramolecular intermediates perceived independently by atomic force microscopy and cryo-electron tomography. We estimated the number of monomeric protein units present in these minute structures. This approach is ideally suited for the investigation of the molecular mechanisms of amyloid formation both in vitro and in the cellular milieu. © 2012 Biophysical Society. Fil:Roberti, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bossi, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Jares-Erijman, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00063495_v102_n7_p1598_Roberti |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
alpha synuclein nanomaterial rhodamine article chemistry color fluorescence microscopy HeLa cell human intracellular space metabolism methodology molecular imaging protein multimerization protein secondary structure alpha-Synuclein Color HeLa Cells Humans Intracellular Space Microscopy, Fluorescence Molecular Imaging Nanostructures Protein Multimerization Protein Structure, Secondary Rhodamines |
| spellingShingle |
alpha synuclein nanomaterial rhodamine article chemistry color fluorescence microscopy HeLa cell human intracellular space metabolism methodology molecular imaging protein multimerization protein secondary structure alpha-Synuclein Color HeLa Cells Humans Intracellular Space Microscopy, Fluorescence Molecular Imaging Nanostructures Protein Multimerization Protein Structure, Secondary Rhodamines Roberti, M.J. Fölling, J. Celej, M.S. Bossi, M. Jovin, T.M. Jares-Erijman, E.A. Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| topic_facet |
alpha synuclein nanomaterial rhodamine article chemistry color fluorescence microscopy HeLa cell human intracellular space metabolism methodology molecular imaging protein multimerization protein secondary structure alpha-Synuclein Color HeLa Cells Humans Intracellular Space Microscopy, Fluorescence Molecular Imaging Nanostructures Protein Multimerization Protein Structure, Secondary Rhodamines |
| description |
The morphological features of α-synuclein (AS) amyloid aggregation in vitro and in cells were elucidated at the nanoscale by far-field subdiffraction fluorescence localization microscopy. Labeling AS with rhodamine spiroamide probes allowed us to image AS fibrillar structures by fluorescence stochastic nanoscopy with an enhanced resolution at least 10-fold higher than that achieved with conventional, diffraction-limited techniques. The implementation of dual-color detection, combined with atomic force microscopy, revealed the propagation of individual fibrils in vitro. In cells, labeled protein appeared as amyloid aggregates of spheroidal morphology and subdiffraction sizes compatible with in vitro supramolecular intermediates perceived independently by atomic force microscopy and cryo-electron tomography. We estimated the number of monomeric protein units present in these minute structures. This approach is ideally suited for the investigation of the molecular mechanisms of amyloid formation both in vitro and in the cellular milieu. © 2012 Biophysical Society. |
| format |
JOUR |
| author |
Roberti, M.J. Fölling, J. Celej, M.S. Bossi, M. Jovin, T.M. Jares-Erijman, E.A. |
| author_facet |
Roberti, M.J. Fölling, J. Celej, M.S. Bossi, M. Jovin, T.M. Jares-Erijman, E.A. |
| author_sort |
Roberti, M.J. |
| title |
Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| title_short |
Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| title_full |
Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| title_fullStr |
Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| title_full_unstemmed |
Imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| title_sort |
imaging nanometer-sized α-synuclein aggregates by superresolution fluorescence localization microscopy |
| url |
http://hdl.handle.net/20.500.12110/paper_00063495_v102_n7_p1598_Roberti |
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