Two-color nanoscopy of three-dimensional volumes by 4Pi detection of stochastically switched fluorophores.
Nat Methods
; 8(4): 353-9, 2011 Apr.
Article
em En
| MEDLINE
| ID: mdl-21399636
ABSTRACT
We demonstrate three-dimensional (3D) super-resolution imaging of stochastically switched fluorophores distributed across whole cells. By evaluating the higher moments of the diffraction spot provided by a 4Pi detection scheme, single markers can be simultaneously localized with <10 nm precision in three dimensions in a layer of 650 nm thickness at an arbitrarily selected depth in the sample. By splitting the fluorescence light into orthogonal polarization states, our 4Pi setup also facilitates the 3D nanoscopy of multiple fluorophores. Offering a combination of multicolor recording, nanoscale resolution and extended axial depth, our method substantially advances the noninvasive 3D imaging of cells and of other transparent materials.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Corantes Fluorescentes
/
Microscopia de Fluorescência
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article