Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera

We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (∼0.2 μm) and temporal (∼50 ps) resolution and allows rapid data acquisition and reliable and reproducible lifetime...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Review of scientific instruments 2003-05, Vol.74 (5), p.2714-2721
Hauptverfasser: Krishnan, R. V., Saitoh, H., Terada, H., Centonze, V. E., Herman, B.
Format: Artikel
Sprache:eng
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung:We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (∼0.2 μm) and temporal (∼50 ps) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in the literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens including stained pollen grains and fibroblast cells expressing green florescent protein. The lifetime values obtained matched well with those reported earlier by other groups for these same specimens. Potential applications of the present system include the measurement of intracellular physiology and fluorescence resonance energy transfer imaging, which are discussed in the context of live cell imaging.
ISSN:0034-6748
1089-7623
DOI:10.1063/1.1569410