Two-photon imaging of multiple fluorescent proteins by phase-shaping and linear unmixing with a single broadband laser.
Opt Express
; 21(14): 17256-64, 2013 Jul 15.
Article
em En
| MEDLINE
| ID: mdl-23938572
ABSTRACT
Imaging multiple fluorescent proteins (FPs) by two-photon microscopy has numerous applications for studying biological processes in thick and live samples. Here we demonstrate a setup utilizing a single broadband laser and a phase-only pulse-shaper to achieve imaging of three FPs (mAmetrine, TagRFPt, and mKate2) in live mammalian cells. Phase-shaping to achieve selective excitation of the FPs in combination with post-imaging linear unmixing enables clean separation of the fluorescence signal of each FP. This setup also benefits from low overall cost and simple optical alignment, enabling easy adaptation in a regular biomedical research laboratory.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Aumento da Imagem
/
Microscopia de Fluorescência por Excitação Multifotônica
/
Lasers
/
Proteínas Luminescentes
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article