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Time-Domain Fluorescence Lifetime Imaging of cAMP Levels with EPAC-Based FRET Sensors.
Kukk, Olga; Klarenbeek, Jeffrey; Jalink, Kees.
Afiliación
  • Kukk O; The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Klarenbeek J; The Netherlands Cancer Institute, Amsterdam, The Netherlands.
  • Jalink K; The Netherlands Cancer Institute, Amsterdam, The Netherlands. k.jalink@nki.nl.
Methods Mol Biol ; 2483: 105-116, 2022.
Article en En | MEDLINE | ID: mdl-35286672
Second messenger molecules in eukaryotic cells relay the signals from activated cell surface receptors to intracellular effector proteins. FRET-based sensors are ideal to visualize and measure the often rapid changes of second messenger concentrations in time and place. Fluorescence Lifetime Imaging (FLIM) is an intrinsically quantitative technique for measuring FRET. Given the recent development of commercially available, sensitive and photon-efficient FLIM instrumentation, it is becoming the method of choice for FRET detection in signaling studies. Here, we describe a detailed protocol for time domain FLIM, using the EPAC-based FRET sensor to measure changes in cellular cAMP levels with high spatiotemporal resolution as an example.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transferencia Resonante de Energía de Fluorescencia / Imagen Óptica Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article País de afiliación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transferencia Resonante de Energía de Fluorescencia / Imagen Óptica Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article País de afiliación: Países Bajos
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