A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging
A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically t...
Guardado en:
| Autores principales: | , , , , , , , , , , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_19327447_v120_n23_p12860_Cusido |
| Aporte de: |
| id |
todo:paper_19327447_v120_n23_p12860_Cusido |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_19327447_v120_n23_p12860_Cusido2023-10-03T16:36:02Z A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging Cusido, J. Ragab, S.S. Thapaliya, E.R. Swaminathan, S. Garcia-Amorós, J. Roberti, M.J. Araoz, B. Mazza, M.M.A. Yamazaki, S. Scott, A.M. Raymo, F.M. Bossi, M.L. Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically the ground-state absorption of the coumarin component. Selective excitation of the photochemical product then produces significant fluorescence and allows the detection of activated bioconjugates at the single-molecule level. Such fluorescence activation events can be exploited to resolve temporally individual emitters and reconstruct images of immunolabeled cells with subdiffraction resolution. Relying on a similar conjugation protocol, a model compound, incorporating the same chromophore of the photochemical product, can also be connected covalently to a secondary antibody. Stimulated emission can be exploited to deplete the excited state of the bioconjugated chromophores and switch their fluorescence off. These operating principles for fluorescence switching also permit the imaging of immunolabeled cells with subdiffraction resolution. Thus, these photoswitchable molecules, in combination with the labeling ability of antibodies, can evolve into valuable probes for bioimaging with superresolution. © 2016 American Chemical Society. Fil:Roberti, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Araoz, B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bossi, M.L. 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_19327447_v120_n23_p12860_Cusido |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies |
| spellingShingle |
Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies Cusido, J. Ragab, S.S. Thapaliya, E.R. Swaminathan, S. Garcia-Amorós, J. Roberti, M.J. Araoz, B. Mazza, M.M.A. Yamazaki, S. Scott, A.M. Raymo, F.M. Bossi, M.L. A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| topic_facet |
Biomolecules Chemical activation Chromophores Excited states Fluorescence Ground state Molecules Nitrogen compounds Optical resolving power Fluorescence switching Ground state absorption Molecular skeleton Operating principles Secondary antibodies Selective excitations Single molecule level Super resolution imaging Antibodies |
| description |
A coumarin fluorophore and an oxazine photochrome can be integrated within the same molecular skeleton and connected covalently to a secondary antibody. Illumination of the antibody-dyad conjugate at an appropriate activation wavelength opens the oxazine ring reversibly and shifts bathochromically the ground-state absorption of the coumarin component. Selective excitation of the photochemical product then produces significant fluorescence and allows the detection of activated bioconjugates at the single-molecule level. Such fluorescence activation events can be exploited to resolve temporally individual emitters and reconstruct images of immunolabeled cells with subdiffraction resolution. Relying on a similar conjugation protocol, a model compound, incorporating the same chromophore of the photochemical product, can also be connected covalently to a secondary antibody. Stimulated emission can be exploited to deplete the excited state of the bioconjugated chromophores and switch their fluorescence off. These operating principles for fluorescence switching also permit the imaging of immunolabeled cells with subdiffraction resolution. Thus, these photoswitchable molecules, in combination with the labeling ability of antibodies, can evolve into valuable probes for bioimaging with superresolution. © 2016 American Chemical Society. |
| format |
JOUR |
| author |
Cusido, J. Ragab, S.S. Thapaliya, E.R. Swaminathan, S. Garcia-Amorós, J. Roberti, M.J. Araoz, B. Mazza, M.M.A. Yamazaki, S. Scott, A.M. Raymo, F.M. Bossi, M.L. |
| author_facet |
Cusido, J. Ragab, S.S. Thapaliya, E.R. Swaminathan, S. Garcia-Amorós, J. Roberti, M.J. Araoz, B. Mazza, M.M.A. Yamazaki, S. Scott, A.M. Raymo, F.M. Bossi, M.L. |
| author_sort |
Cusido, J. |
| title |
A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| title_short |
A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| title_full |
A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| title_fullStr |
A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| title_full_unstemmed |
A Photochromic Bioconjugate with Photoactivatable Fluorescence for Superresolution Imaging |
| title_sort |
photochromic bioconjugate with photoactivatable fluorescence for superresolution imaging |
| url |
http://hdl.handle.net/20.500.12110/paper_19327447_v120_n23_p12860_Cusido |
| work_keys_str_mv |
AT cusidoj aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT ragabss aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT thapaliyaer aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT swaminathans aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT garciaamorosj aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT robertimj aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT araozb aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT mazzamma aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT yamazakis aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT scottam aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT raymofm aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT bossiml aphotochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT cusidoj photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT ragabss photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT thapaliyaer photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT swaminathans photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT garciaamorosj photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT robertimj photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT araozb photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT mazzamma photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT yamazakis photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT scottam photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT raymofm photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging AT bossiml photochromicbioconjugatewithphotoactivatablefluorescenceforsuperresolutionimaging |
| _version_ |
1807320544480067584 |