Photostable and Orthogonal Solvatochromic Fluorophores for Simultaneous In Situ Quantification of Multiple Cellular Signaling Molecules.
ACS Chem Biol
; 15(7): 1913-1920, 2020 07 17.
Article
em En
| MEDLINE
| ID: mdl-32525312
ABSTRACT
Ratiometric fluorescence sensors are powerful tools for direct quantification of diverse biological analytes. To overcome a shortage of solvatochromic fluorophores crucial for in situ ratiometric imaging of biological targets, we prepared and characterized a small library of modular fluorophores with diverse spectral properties. Among them, WCB and WCR showed excellent spectral properties, including high photostability, brightness, and solvatochromism, and are ideally suited for dual ratiometric imaging due to their spectral orthogonality. By conjugating WCB and WCR with protein-based lipid sensors, we were able to achieve robust simultaneous in situ quantitative imaging of two metabolically linked signaling lipids, phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate in live cells. This study shows that any combination of signaling molecules can be simultaneously quantified in a spatiotemporally resolved manner by ratiometric imaging with finely tuned solvatochromic fluorophores.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Fosfatos de Fosfatidilinositol
/
Fluorenos
/
Corantes Fluorescentes
Limite:
Animals
Idioma:
En
Revista:
ACS Chem Biol
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Estados Unidos